U.S. patent application number 16/020558 was filed with the patent office on 2019-06-06 for cross-species-specific psmaxcd3 bispecific single chain antibody.
The applicant listed for this patent is AMGEN RESEARCH (MUNICH) GMBH. Invention is credited to Petra Fluhr, Susanne Hausmann, Patrick Hoffmann, Roman Kischel, Matthias Klinger, Peter Kufer, Ralf Lutterbuse, Susanne Mangold, Doris Rau, Tobias Raum, Evelyne Schaller, Carola Steiger.
Application Number | 20190169310 16/020558 |
Document ID | / |
Family ID | 42073956 |
Filed Date | 2019-06-06 |
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United States Patent
Application |
20190169310 |
Kind Code |
A1 |
Kufer; Peter ; et
al. |
June 6, 2019 |
CROSS-SPECIES-SPECIFIC PSMAxCD3 BISPECIFIC SINGLE CHAIN
ANTIBODY
Abstract
The present invention relates to a bispecific single chain
antibody molecule comprising a first binding domain capable of
binding to an epitope of human and non-chimpanzee primate CD3
epsilon chain, wherein the epitope is part of an amino acid
sequence comprised in the group consisting of SEQ ID NOs. 2, 4, 6,
and 8, and a second binding domain capable of binding to
prostate-specific membrane antigen (PSMA). The invention also
provides nucleic acids encoding said bispecific single chain
antibody molecule as well as vectors and host cells and a process
for its production. The invention further relates to pharmaceutical
compositions comprising said bispecific single chain antibody
molecule and medical uses of said bispecific single chain anti-body
molecule.
Inventors: |
Kufer; Peter; (Munich,
DE) ; Raum; Tobias; (Munich, DE) ; Kischel;
Roman; (Munich, DE) ; Lutterbuse; Ralf;
(Munich, DE) ; Hoffmann; Patrick; (Munich, DE)
; Rau; Doris; (Munich, DE) ; Mangold; Susanne;
(Munich, DE) ; Klinger; Matthias; (Munich, DE)
; Schaller; Evelyne; (Munich, DE) ; Hausmann;
Susanne; (Munich, DE) ; Fluhr; Petra; (Munich,
DE) ; Steiger; Carola; (Munich, DE) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
AMGEN RESEARCH (MUNICH) GMBH |
Munich |
|
DE |
|
|
Family ID: |
42073956 |
Appl. No.: |
16/020558 |
Filed: |
June 27, 2018 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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13122245 |
Jul 15, 2011 |
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PCT/EP2009/062793 |
Oct 1, 2009 |
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16020558 |
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61101857 |
Oct 1, 2008 |
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Current U.S.
Class: |
1/1 |
Current CPC
Class: |
C07K 16/3069 20130101;
C07K 2317/31 20130101; A61K 2039/505 20130101; C07K 2317/34
20130101; A61P 37/00 20180101; A61P 13/08 20180101; A61P 35/00
20180101 |
International
Class: |
C07K 16/30 20060101
C07K016/30 |
Claims
1. A bispecific single chain antibody molecule comprising a first
binding domain capable of binding to an epitope of human CD3epsilon
(CD3.epsilon.) chain and non-chimpanzee primate-CD3.epsilon. chain,
wherein the epitope is part of the amino acid sequence set forth in
SEQ ID NO: 2, 4, 6, or 8, and comprises at least the amino sequence
Gln-Asp-Gly-Asn-Glu (QDGNE) (SEQ ID NO: 1050), and a second binding
domain capable of binding to prostate-specific membrane antigen
(PSMA).
2. The bispecific single chain antibody molecule of claim 1,
wherein at least one of said first or second binding domain is
CDR-grafted, humanized or human.
3. The bispecific single chain antibody molecule according to claim
1, wherein the first binding domain comprises a VL region selected
from the group consisting of: (a) a VL region comprising a CDR-L1
as depicted in SEQ ID NO: 27, a CDR-L2 as depicted in SEQ ID NO:
28, and a CDR-L3 as depicted in SEQ ID NO: 29; (b) a VL region
comprising a CDR-L1 as depicted in SEQ ID NO: 117, a CDR-L2 as
depicted in SEQ ID NO: 118, and a CDR-L3 as depicted in SEQ ID NO:
119; and (c) a VL region comprising a CDR-L1 as depicted in SEQ ID
NO: 153, a CDR-L2 as depicted in SEQ ID NO: 154, and a CDR-L3 as
depicted in SEQ ID NO: 155.
4. The bispecific single chain antibody molecule according to claim
1, wherein the first binding domain comprises a VH region selected
from the group consisting of: (a) a VH region comprising a CDR-H1
as depicted in SEQ ID NO: 12, a CDR-H2 as depicted in SEQ ID NO:
13, and a CDR-H3 as depicted in SEQ ID NO: 14; (b) a VH region
comprising a CDR-H1 as depicted in SEQ ID NO: 30, a CDR-H2 as
depicted in SEQ ID NO: 31, and a CDR-H3 as depicted in SEQ ID NO:
32; (c) a VH region comprising a CDR-H1 as depicted in SEQ ID NO:
48, a CDR-H2 as depicted in SEQ ID NO: 49.sub.1 and a CDR-H3 as
depicted in SEQ ID NO: 50; (d) a VH region comprising a CDR-H1 as
depicted in SEQ ID NO: 66, a CDR-H2 as depicted in SEQ ID NO: 67,
and a CDR-H3 as depicted in SEQ ID NO: 68; (e) a VH region
comprising a CDR-H1 as depicted in SEQ ID NO: 84, a CDR-H2 as
depicted in SEQ ID NO: 85, and a CDR-H3 as depicted in SEQ ID NO:
86; (f) a VH region comprising a CDR-H1 as depicted in SEQ ID NO:
102, a CDR-H2 as depicted in SEQ ID NO: 103, and a CDR-H3 as
depicted in SEQ ID NO: 104; (g) a VH region comprising a CDR-H1 as
depicted in SEQ ID NO: 120, a CDR-H2 as depicted in SEQ ID NO: 121,
and a CDR-H3 as depicted in SEQ ID NO: 122; (h) a VH region
comprising a CDR-H1 as depicted in SEQ ID NO: 138, a CDR-H2 as
depicted in SEQ ID NO: 139, and a CDR-H3 as depicted in SEQ ID NO:
140; (i) a VH region comprising a CDR-H1 as depicted in SEQ ID NO:
156, a CDR-H2 as depicted in SEQ ID NO: 157, and a CDR-H3 as
depicted in SEQ ID NO: 158; and (j) a VH region comprising a CDR-H1
as depicted in SEQ ID NO: 174, a CDR-H2 as depicted in SEQ ID NO:
175, and a CDR-H3 as depicted in SEQ ID NO: 176.
5. The bispecific single chain antibody molecule according to claim
1, wherein the first binding domain comprises a VL region as
depicted in SEQ ID NO: 35, 39, 125, 129, 161 or 165.
6. The bispecific single chain antibody molecule according to claim
1, wherein the first binding domain comprises a VH region as
depicted in SEQ ID NO: 15, 19, 33, 37, 51, 55, 69, 73, 87, 91, 105,
109, 123, 127, 141, 145, 159, 163, 177 or 181.
7. The bispecific single chain antibody molecule according to claim
1, wherein the first binding domain is selected from the group
consisting of: (a) a first binding domain comprising a VL region as
depicted in SEQ ID NO: 17 or 21 and a VH region as depicted in SEQ
ID NO: 15 or 19; (b) a first binding domain comprising a VL region
as depicted in SEQ ID NO: 35 or 39 and a VH region as depicted in
SEQ ID NO: 33 or 37; (c) a first binding domain comprising a VL
region as depicted in SEQ ID NO: 53 or 57 and a VH region as
depicted in SEQ ID NO: 51 or 55; (d) a first binding domain
comprising a VL region as depicted in SEQ ID NO: 71 or 75 and a VH
region as depicted in SEQ ID NO: 69 or 73; (e) a first binding
domain comprising a VL region as depicted in SEQ ID NO: 89 or 93
and a VH region as depicted in SEQ ID NO: 87 or 91; (f) a first
binding domain comprising a VL region as depicted in SEQ ID NO: 107
or 111 and a VH region as depicted in SEQ ID NO: 105 or 109; (g) a
first binding domain comprising a VL region as depicted in SEQ ID
NO: 125 or 129 and a VH region as depicted in SEQ ID NO: 123 or
127; (h) a first binding domain comprising a VL region as depicted
in SEQ ID NO: 143 or 147 and a VH region as depicted in SEQ ID NO:
141 or 145; (i) a first binding domain comprising a VL region as
depicted in SEQ ID NO: 161 or 165 and a VH region as depicted in
SEQ ID NO: 159 or 163; and (j) a first binding domain comprising a
VL region as depicted in SEQ ID NO: 179 or 183 and a VH region as
depicted in SEQ ID NO: 177 or 181.
8. The bispecific single chain antibody molecule according to claim
1, wherein the first binding domain comprises the amino acid
sequence set forth in SEQ ID NO: 23, 25, 41, 43, 59, 61, 77, 79,
95, 97, 113, 115, 131, 133, 149, 151, 167, 169, 185 or 187.
9. The bispecific single chain antibody molecule according to claim
1, wherein the PSMA is human PSMA and/or a non-chimpanzee primate
PSMA.
10. The bispecific single chain antibody molecule according to
claim 9, wherein the the second binding domain is selected from the
group consisting of: a) a second binding domain comprising CDR H1-3
of SEQ ID NO: 394-396 and CDR L1-3 of SEQ ID NO: 389-391; b) a
second binding domain comprising CDR H1-3 of SEQ ID NO: 408-410 and
CDR L1-3 of SEQ ID NO: 403-405; c) a second binding domain
comprising CDR H1-3 of SEQ ID NO: 422-424 and CDR L1-3 of SEQ ID
NO: 417-419; d) a second binding domain comprising CDR H1-3 of SEQ
ID NO: 436-438 and CDR L1-3 of SEQ ID NO: 431-433; e) a second
binding domain comprising CDR H1-3 of SEQ ID NO: 445-447 and CDR
L1-3 of SEQ ID NO: 450-452; f) a second binding domain comprising
CDR H1-3 of SEQ ID NO: 464-466 and CDR L1-3 of SEQ ID NO: 459-461;
g) a second binding domain comprising CDR H1-3 of SEQ ID NO:
478-480 and CDR L1-3 of SEQ ID NO: 473-475; h) a second binding
domain comprising CDR H1-3 of SEQ ID NO: 492-494 and CDR L1-3 of
SEQ ID NO: 487-489; i) a second binding domain comprising CDR H1-3
of SEQ ID NO: 506-508 and CDR L1-3 of SEQ ID NO: 501-503; j) a
second binding domain comprising CDR H1-3 of SEQ ID NO: 520-522 and
CDR L1-3 of SEQ ID NO: 515-517; k) a second binding domain
comprising CDR H1-3 of SEQ ID NO: 534-536 and CDR L1-3 of SEQ ID
NO: 529-531; l) a second binding domain comprising CDR H1-3 of SEQ
ID NO: 548-550 and CDR L1-3of SEQ ID NO: 543-545; m) a second
binding domain comprising CDR H1-3 of SEQ ID NO: 562-564 and CDR
L1-3 of SEQ ID NO: 557-559; n) a second binding domain comprising
CDR H1-3 of SEQ ID NO: 576-578 and CDR L1-3 of SEQ ID NO: 571-573;
o) a second binding domain comprising CDR H1-3 of SEQ ID NO:
590-592 and CDR L1-3 of SEQ ID NO: 585-587; p) a second binding
domain comprising CDR H1-3 of SEQ ID NO: 604-606 and CDR L1-3 of
SEQ ID NO: 599-601; q) a second binding domain comprising CDR H1-3
of SEQ ID NO: 618-620 and CDR L1-3 of SEQ ID NO: 613-615; r) a
second binding domain comprising CDR H1-3 of SEQ ID NO: 632-634 and
CDR L1-3 of SEQ ID NO: 627-629; s) a second binding domain
comprising CDR H1-3 of SEQ ID NO: 646-648 and CDR L1-3 of SEQ ID
NO: 641-643; t) a second binding domain comprising CDR H1-3 of SEQ
ID NO: 660-662 and CDR L1-3 of SEQ ID NO: 655-657; u) a second
binding domain comprising CDR H1-3 of SEQ ID NO: 674-676 and CDR
L1-3 of SEQ ID NO: 669-671; v) a second binding domain comprising
CDR H1-3 of SEQ ID NO: 688-690 and CDR L1-3 of SEQ ID NO: 683-685;
w) a second binding domain comprising CDR H1-3 of SEQ ID NO:
702-704 and CDR L1-3 of SEQ ID NO: 697-699; x) a second binding
domain comprising CDR H1-3 of SEQ ID NO: 716-718 and CDR L1-3 of
SEQ ID NO: 711-713; y) a second binding domain comprising CDR H1-3
of SEQ ID NO: 729-731 and CDR L1-3 of SEQ ID NO: 724-726; z) a
second binding domain comprising CDR H1-3 of SEQ ID NO: 788-790 and
CDR L1-3 of SEQ ID NO: 793-795; aa) a second binding domain
comprising CDR H1-3 of SEQ ID NO: 806-808 and CDR L1-3 of SEQ ID
NO: 811-813; ab) a second binding domain comprising CDR H1-3 of SEQ
ID NO: 852-854 and CDR L1-3 of SEQ ID NO: 857-859; ac) a second
binding domain comprising CDR H1-3 of SEQ ID NO: 838-840 and CDR
L1-3 of SEQ ID NO: 843-845; ad) a second binding domain comprising
CDR H1-3 of SEQ ID NO: 824-826 and CDR L1-3 of SEQ ID NO: 829-831;
ae) a second binding domain comprising CDR H1-3 of SEQ ID NO:
774-776 and CDR L1-3 of SEQ ID NO: 779-781; af) a second binding
domain comprising CDR H1-3 of SEQ ID NO: 688-690 and CDR L1-3 of
SEQ ID NO. 683-685; ag) a second binding domain comprising CDR H1-3
of SEQ ID NO: 870-872 and CDR L1-3 of SEQ ID NO: 875-877; ah) a
second binding domain comprising CDR H1-3 of SEQ ID NO: 888-890 and
CDR L1-3 of SEQ ID NO: 893-895; ai) a second binding domain
comprising CDR H1-3 of SEQ ID NO: 924-926 and CDR L1-3 of SEQ ID
NO: 929-931; aj) a second binding domain comprising CDR H1-3 of SEQ
ID NO: 1019-1021 and CDR L1-3 of SEQ ID NO: 1025-1027; ak) a second
binding domain comprising CDR H1-3 of SEQ ID NO: 1006-1008 and CDR
L1-3 of SEQ ID NO: 1011-1013; al) a second binding domain
comprising CDR H1-3 of SEQ ID NO: 906-908 and CDR L1-3 of SEQ ID
NO: 911-913; am) a second binding domain comprising CDR H1-3 of SEQ
ID NO: 992-994 and CDR L1-3 of SEQ ID NO: 997-999; an) a second
binding domain comprising CDR H1-3 of SEQ ID NO: 942-944 and CDR
L1-3 of SEQ ID NO: 947-949; ao) a second binding domain comprising
CDR H1-3 of SEQ ID NO: 960-962 and CDR L1-3 of SEQ ID NO: 965-967;
and ap) a second binding domain comprising CDR H1-3 of SEQ ID NO:
978-980 and CDR L1-3 of SEQ ID NO: 983-985.
11. The bispecific single chain antibody molecule of claim 10,
wherein the binding domains are arranged in the order VH PSMA-VL
PSMA-VH CD3-VL CD3 or VL PSMA-VH CD3-VL CD3.
12. The bispecific single chain antibody molecule according to
claim 11, wherein the bispecific single chain antibody molecule
comprises an amino acid sequence selected from the group consisting
of: (a) an amino acid sequence as depicted in any of SEQ ID NO:
399, 413, 427, 441, 455, 469, 483, 497, 511, 525, 539, 553, 567,
581, 595, 609, 623, 637, 651, 665, 679, 693, 707, 721, 734, 799,
817, 863, 849, 835, 785, 899, 935, 1017, 1031, 917, 1003, 953, 971,
or 989; (b) an amino acid sequence encoded by a nucleic acid
sequence as depicted in any of SEQ ID NO: 400, 414, 428, 442, 456,
470, 484, 498, 512, 526, 540, 554, 568, 582, 596, 610, 624, 638,
652, 666, 680, 694, 708, 736, 735, 818, 864, 850, 836, 786, 882,
900, 936, 1018, 1032, 918, 1004, 954, 972, 990, 804, 822, 868, 886,
904, 940, 922, 958, or 976; and (c) an amino acid sequence at least
90% identical to the amino acid sequence of (a) or (b).
13. A nucleic acid comprising a nucleotide sequence encoding the
bispecific single chain antibody molecule of claim 1.
14. A vector comprising the nucleic acid of claim 13.
15-16. (canceled)
17. A host transformed or transfected with the vector of claim
14.
18. A process for producing a bispecific single chain antibody
molecule comprising culturing the host of claim 17 under conditions
suitable to express the bispecific single chain antibody molecule
and, optionally recovering the bispecific single chain antibody
molecule from the culture.
19. A pharmaceutical composition comprising the bispecific single
chain antibody molecule according to claim 1.
20-27. (canceled)
28. A method for preventing, treating, or ameliorating a disease in
a subject in the need thereof, said method comprising the step of
administering an effective amount of the pharmaceutical composition
of claim 19.
29. The method of claim 28, wherein said disease is cancer.
30. The method or use of claim 29, wherein said cancer is a solid
tumor, preferably a carcinoma or prostate cancer.
31. The method of claim 28, wherein said pharmaceutical composition
is administered in combination with an additional drug.
32-33. (canceled)
34. The method of claim 28, wherein said subject is a human.
35. A kit comprising the bispecific single chain antibody molecule
of claim 1.
Description
[0001] The present invention relates to a bispecific single chain
antibody molecule comprising a first binding domain capable of
binding to an epitope of human and non-chimpanzee primate CD3
epsilon chain, wherein the epitope is part of an amino acid
sequence comprised in the group consisting of SEQ ID NOs. 2, 4, 6,
and 8, and a second binding domain capable of binding to
prostate-specific membrane antigen (PSMA). The invention also
provides nucleic acids encoding said bispecific single chain
antibody molecule as well as vectors and host cells and a process
for its production. The invention further relates to pharmaceutical
compositions comprising said bispecific single chain antibody
molecule and medical uses of said bispecific single chain antibody
molecule.
[0002] T cell recognition is mediated by clonotypically distributed
alpha beta and gamma delta T cell receptors (TcR) that interact
with the peptide-loaded molecules of the peptide MHC (pMHC) (Davis
& Bjorkman, Nature 334 (1988), 395-402). The antigen-specific
chains of the TcR do not possess signalling domains but instead are
coupled to the conserved multisubunit signaling apparatus CD3
(Call, Cell 111 (2002), 967-979, Alarcon, Immunol. Rev. 191 (2003),
38-46, Malissen Immunol. Rev. 191 (2003), 7-27). The mechanism by
which TcR ligation is directly communicated to the signalling
apparatus remains a fundamental question in T cell biology
(Alarcon, loc. cit.; Davis, Cell 110 (2002), 285-287). It seems
clear that sustained T cell responses involve coreceptor
engagement, TcR oligomerization, and a higher order arrangement of
TcR-pMHC complexes in the immunological synapse (Davis & van
der Merwe, Curr. Biol. 11 (2001), R289-R291, Davis, Nat. Immunol. 4
(2003), 217-224). However very early TcR signalling occurs in the
absence of these events and may involve a ligand-induced
conformational change in CD3 epsilon (Alarcon, loc. cit., Davis
(2002), loc. cit., Gil, J. Biol. Chem. 276 (2001), 11174-11179,
Gil, Cell 109 (2002), 901-912). The epsilon, gamma, delta and zeta
subunits of the signaling complex associate with each other to form
a CD3 epsilon-gamma heterodimer, a CD3 epsilon-delta
.quadrature.heterodimer, and a CD3 zeta-zeta homodimer (Call, loc.
cit.). Various studies have revealed that the CD3 molecules are
important for the proper cell surface expression of the alpha beta
TcR and normal T cell development (Berkhout, J. Biol. Chem. 263
(1988), 8528-8536, Wang, J. Exp. Med. 188 (1998), 1375-1380,
Kappes, Curr. Opin. Immunol. 7 (1995), 441-447). The solution
structure of the ectodomain fragments of the mouse CD3 epsilon
gamma heterodimer showed that the epsilon gamma subunits are both
C2-set Ig domains that interact with each other to form an unusual
side-to-side dimer configuration (Sun, Cell 105 (2001), 913-923).
Although the cysteine-rich stalk appears to play an important role
in driving CD3 dimerization (Su, loc. cit., Borroto, J. Biol. Chem.
273 (1998), 12807-12816), interaction by means of the extracellular
domains of CD3 epsilon and CD3 gamma is sufficient for assembly of
these proteins with TcR beta (Manolios, Eur. J. Immunol. 24 (1994),
84-92, Manolios & Li, Immunol. Cell Biol. 73 (1995), 532-536).
Although still controversial, the dominant stoichiometry of the TcR
most likely comprises one alpha beta TcR, one CD3 epsilon gamma
heterodimer, one CD3 epsilon delta heterodimer and one CD3 zeta
zeta homodimer (Call, loc. cit.). Given the central role of the
human CD3 epsilon gamma heterodimer in the immune response, the
crystal structure of this complex bound to the therapeutic antibody
OKT3 has recently been elucidated (Kjer-Nielsen, PNAS 101, (2004),
7675-7680).
[0003] A number of therapeutic strategies modulate T cell immunity
by targeting TcR signaling, particularly the anti-human CD3
monoclonal antibodies (mAbs) that are widely used clinically in
immunosuppressive regimes. The CD3-specific mouse mAb OKT3 was the
first mAb licensed for use in humans (Sgro, Toxicology 105 (1995),
23-29) and is widely used clinically as an immunosuppressive agent
in transplantation (Chatenoud, Clin. Transplant 7 (1993), 422-430,
Chatenoud, Nat. Rev. Immunol. 3 (2003), 123-132, Kumar, Transplant.
Proc. 30 (1998), 1351-1352), type 1 diabetes (Chatenoud (2003),
loc. cit.), and psoriasis (Utset, J. Rheumatol. 29 (2002),
1907-1913). Moreover, anti-CD3 mAbs can induce partial T cell
signalling and clonal anergy (Smith, J. Exp. Med. 185 (1997),
1413-1422). OKT3 has been described in the literature as a potent T
cell mitogen (Van Wauve, J. Immunol. 124 (1980), 2708-18) as well
as a potent T cell killer (Wong, Transplantation 50 (1990), 683-9).
OKT3 exhibits both of these activities in a time-dependent fashion;
following early activation of T cells leading to cytokine release,
upon further administration OKT3 later blocks all known T cell
functions. It is due to this later blocking of T cell function that
OKT3 has found such wide application as an immunosuppressant in
therapy regimens for reduction or even abolition of allograft
tissue rejection.
[0004] OKT3 reverses allograft tissue rejection most probably by
blocking the function of all T cells, which play a major role in
acute rejection. OKT3 reacts with and blocks the function of the
CD3 complex in the membrane of human T cells, which is associated
with the antigen recognition structure of T cells (TCR) and is
essential for signal transduction. Which subunit of the TCR/CD3 is
bound by OKT3 has been the subject of multiple studies. Though some
evidence has pointed to a specificity of OKT3 for the
epsilon-subunit of the TCR/CD3 complex (Tunnacliffe, Int. Immunol.
1 (1989), 546-50; Kjer-Nielsen, PNAS 101, (2004), 7675-7680).
Further evidence has shown that OKT3 binding of the TCR/CD3 complex
requires other subunits of this complex to be present (Salmeron, J.
Immunol. 147 (1991), 3047-52).
[0005] Other well known antibodies specific for the CD3 molecule
are listed in Tunnacliffe, Int. Immunol. 1 (1989), 546-50. As
indicated above, such CD3 specific antibodies are able to induce
various T cell responses such as lymphokine production (Von Wussow,
J. Immunol. 127 (1981), 1197; Palacious, J. Immunol. 128 (1982),
337), proliferation (Van Wauve, J. Immunol. 124 (1980), 2708-18)
and suppressor-T cell induction (Kunicka, in "Lymphocyte Typing II"
1 (1986), 223). That is, depending on the experimental conditions,
CD3 specific monoclonal antibody can either inhibit or induce
cytotoxicity (Leewenberg, J. Immunol. 134 (1985), 3770; Phillips,
J. Immunol. 136 (1986) 1579; Platsoucas, Proc. Natl. Acad. Sci. USA
78 (1981), 4500; Itoh, Cell. Immunol. 108 (1987), 283-96; Mentzer,
J. Immunol. 135 (1985), 34; Landegren, J. Exp. Med. 155 (1982),
1579; Choi (2001), Eur. J. Immunol. 31, 94-106; Xu (2000), Cell
Immunol. 200, 16-26; Kimball (1995), Transpl. Immunol. 3,
212-221).
[0006] Although many of the CD3 antibodies described in the art
have been reported to recognize the CD3 epsilon subunit of the CD3
complex, most of them bind in fact to conformational epitopes and,
thus, only recognize CD3 epsilon in the native context of the TCR.
Conformational epitopes are characterized by the presence of two or
more discrete amino acid residues which are separated in the
primary sequence, but come together on the surface of the molecule
when the polypeptide folds into the native protein/antigen (Sela,
(1969) Science 166, 1365 and Laver, (1990) Cell 61, 553-6). The
conformational epitopes bound by CD3 epsilon antibodies described
in the art may be separated in two groups. In the major group, said
epitopes are being formed by two CD3 subunits, e.g. of the CD3
epsilon chain and the CD3 gamma or CD3 delta chain. For example, it
has been found in several studies that the most widely used CD3
epsilon monoclonal antibodies OKT3, WT31, UCHT1, 7D6 and Leu-4 did
not bind to cells singly transfected with the CD3-epsilon chain.
However, these antibodies stained cells doubly transfected with a
combination of CD3 epsilon plus either CD3 gamma or CD3 delta
(Tunnacliffe, loc. cit.; Law, Int. Immunol. 14 (2002), 389-400;
Salmeron, J. Immunol. 147 (1991), 3047-52; Coulie, Eur. J. Immunol.
21 (1991), 1703-9). In a second smaller group, the conformational
epitope is being formed within the CD3 epsilon subunit itself. A
member of this group is for instance mAb APA 1/1 which has been
raised against denatured CD3 epsilon (Risueno, Blood 106 (2005),
601-8). Taken together, most of the CD3 epsilon antibodies
described in the art recognize conformational epitopes located on
two or more subunits of CD3. The discrete amino acid residues
forming the three-dimensional structure of these epitopes may
hereby be located either on the CD3 epsilon subunit itself or on
the CD3 epsilon subunit and other CD3 subunits such as CD3 gamma or
CD3 delta.
[0007] Another problem with respect to CD3 antibodies is that many
CD3 antibodies have been found to be species-specific. Anti-CD3
monoclonal antibodies--as holds true generally for any other
monoclonal antibodies--function by way of highly specific
recognition of their target molecules. They recognize only a single
site, or epitope, on their target CD3 molecule. For example, one of
the most widely used and best characterized monoclonal antibodies
specific for the CD3 complex is OKT-3. This antibody reacts with
chimpanzee CD3 but not with the CD3 homolog of other primates, such
as macaques, or with dog CD3 (Sandusky et al., J. Med. Primatol. 15
(1986), 441-451). Similarly, WO2005/118635 or WO2007/033230
describe human monoclonal CD3 epsilon antibodies which react with
human CD3 epsilon but not with CD3 epsilon of mouse, rat, rabbit or
non-chimpanzee primates such as rhesus monkey, cynomolgus monkey or
baboon monkey. The anti-CD3 monoclonal antibody UCHT-1 is also
reactive with CD3 from chimpanzee but not with CD3 from macaques
(own data). On the other hand, there are also examples of
monoclonal antibodies, which recognize macaque antigens, but not
their human counterparts. One example of this group is monoclonal
antibody FN-18 directed to CD3 from macaques (Uda et al., J. Med.
Primatol. 30 (2001), 141-147). Interestingly, it has been found
that peripheral lymphocytes from about 12% of cynomolgus monkeys
lacked reactivity with anti-rhesus monkey CD3 monoclonal antibody
(FN-18) due to a polymorphism of the CD3 antigen in macaques. Uda
et al. described a substitution of two amino acids in the CD3
sequence of cynomolgus monkeys, which are not reactive with FN-18
antibodies, as compared to CD3 derived from animals, which are
reactive with FN-18 antibodies (Uda et al., J Med Primatol. 32
(2003), 105-10; Uda et al., J Med Primatol. 33 (2004), 34-7).
[0008] The discriminatory ability, i.e. the species specificity,
inherent not only to CD3 monoclonal antibodies (and fragments
thereof), but to monoclonal antibodies in general, is a significant
impediment to their development as therapeutic agents for the
treatment of human diseases. In order to obtain market approval any
new candidate medication must pass through rigorous testing. This
testing can be subdivided into preclinical and clinical phases:
Whereas the latter--further subdivided into the generally known
clinical phases I, II and III--is performed in human patients, the
former is performed in animals. The aim of pre-clinical testing is
to prove that the drug candidate has the desired activity and most
importantly is safe. Only when the safety in animals and possible
effectiveness of the drug candidate has been established in
preclinical testing this drug candidate will be approved for
clinical testing in humans by the respective regulatory authority.
Drug candidates can be tested for safety in animals in the
following three ways, (i) in a relevant species, i.e. a species
where the drug candidates can recognize the ortholog antigens, (ii)
in a transgenic animal containing the human antigens and (iii) by
use of a surrogate for the drug candidate that can bind the
ortholog antigens present in the animal. Limitations of transgenic
animals are that this technology is typically limited to rodents.
Between rodents and man there are significant differences in the
physiology and the safety results cannot be easily extrapolated to
humans. The limitations of a surrogate for the drug candidate are
the different composition of matter compared to the actual drug
candidate and often the animals used are rodents with the
limitation as discussed above. Therefore, preclinical data
generated in rodents are of limited predictive power with respect
to the drug candidate. The approach of choice for safety testing is
the use of a relevant species, preferably a lower primate. The
limitation now of monoclonal antibodies suitable for therapeutic
intervention in man described in the art is that the relevant
species are higher primates, in particular chimpanzees. Chimpanzees
are considered as endangered species and due to their human-like
nature, the use of such animals for drug safety testing has been
banned in Europe and is highly restricted elsewhere. CD3 has also
been successfully used as a target for bispecific single chain
antibodies in order to redirect cytotoxic T cells to pathological
cells, resulting in the depletion of the diseased cells from the
respective organism (WO 99/54440; WO 04/106380). For example,
Bargou et al. (Science 321 (2008):974-7) have recently reported on
the clinical activity of a CD19.times.CD3 bispecific antibody
construct called blinatumomab, which has the potential to engage
all cytotoxic T cells in human patients for lysis of cancer cells.
Doses as low as 0.005 milligrams per square meter per day in
non-Hodgkin's lymphoma patients led to an elimination of target
cells in blood. Partial and complete tumor regressions were first
observed at a dose level of 0.015 milligrams, and all seven
patients treated at a dose level of 0.06 milligrams experienced a
tumor regression. Blinatumomab also led to clearance of tumor cells
from bone marrow and liver. Though this study established clinical
proof of concept for the therapeutic potency of the bispecific
single chain antibody format in treating blood-cell derived cancer,
there is still need for successful concepts for therapies of other
cancer types.
[0009] In 2008, an estimated 186,320 men will be newly diagnosed
with prostate cancer in the United States and about 28,660 men will
die from the disease. The most recent report available on cancer
mortality shows that, in 2004, the overall death rate from prostate
cancer among American men was 25 per 100,000. In the late 1980s,
the widespread adoption of the prostate-specific antigen (PSA) test
represented a major improvement in the management of prostate
cancer. This test measures the amount of PSA protein in the blood,
which is often elevated in patients with prostate cancer. In 1986,
the U.S. Food and Drug Administration approved the use of the PSA
test to monitor patients with prostate cancer and, in 1994,
additionally approved its use as a screening test for this disease.
Due to the widespread implementation of PSA testing in the United
States, approximately 90 percent of all prostate cancers are
currently diagnosed at an early stage, and, consequently, men are
surviving longer after diagnosis. However, the results of two
ongoing clinical trials, the NCI-sponsored Prostate, Lung,
Colorectal, and Ovarian (PLCO) screening trial and the European
Study of Screening for Prostate Cancer (ERSPC) will be needed to
determine whether PSA screening actually saves lives. Ongoing
clinical trials over the past 25 years have investigated the
effectiveness of natural and synthetic compounds in the prevention
of prostate cancer. For example, the Prostate Cancer Prevention
Trial (PCPT), which enrolled nearly 19,000 healthy men, found that
finasteride, a drug approved for the treatment of benign prostatic
hyperplasia (BPH), which is a noncancerous enlargement of the
prostate, reduced the risk of developing prostate cancer by 25
percent. Another trial, the Selenium and Vitamin E Cancer
Prevention Trial (SELECT), is studying more than 35,000 men to
determine whether daily supplements of selenium and vitamin E can
reduce the incidence of prostate cancer in healthy men. Other
prostate cancer prevention trials are currently evaluating the
protective potential of multivitamins, vitamins C and D, soy, green
tea, and lycopene, which is a natural compound found in tomatoes.
One study, reported in 2005, showed that specific genes were fused
in 60 to 80 percent of the prostate tumors analyzed. This study
represents the first observation of non-random gene rearrangements
in prostate cancer. This genetic alteration may eventually be used
as a biomarker to aid in the diagnosis and, possibly, treatment of
this disease. Other studies have shown that genetic variations in a
specific region of chromosome 8 can increase a man's risk of
developing prostate cancer. These genetic variations account for
approximately 25 percent of the prostate cancers that occur in
white men. They are the first validated genetic variants that
increase the risk of developing prostate cancer and may help
scientists better understand the genetic causes of this disease.
There is also ongoing research that examines how proteins
circulating in a patient's blood can be used to improve the
diagnosis of prostate and other cancers. In 2005, scientists
identified a group of specific proteins that are produced by a
patient's immune system in response to prostate tumors. These
proteins, a type of autoantibody, were able to detect the presence
of prostate cancer cells in blood specimens with greater than 90
percent accuracy. When used in combination with PSA, these and
other blood proteins may eventually be used to reduce the number of
false-positive results obtained with PSA testing alone and,
therefore, reduce the large number of unnecessary prostate biopsies
that are performed each year due to false-positive PSA test
results.
[0010] Apart from PSA, several other markers for prostate cancer
have been identified, including e.g. the six-transmembrane
epithelial antigen of the prostate (STEAP) (Hubert et al., PNAS 96
(1999), 14523-14528), the prostate stem cell antigen (PSCA) (Reiter
et al., Proc. Nat. Acad. Sci. 95: 1735-1740, 1998) and the
prostate-specific membrane antigen (PSMA; PSM) (Israeli et al.,
Cancer Res. 53 (1993). PSMA was originally defined by the
monoclonal antibody (MAb) 7E11 derived from immunization with a
partially purified membrane preparation from the lymph node
prostatic adenocarcinoma (LNCaP) cell line (Horoszewicz et al.,
Anticancer Res. 7 (1987), 927-35). A 2.65-kb cDNA fragment encoding
the PSMA protein was cloned and subsequently mapped to chromosome
11p11.2 (Israeli et al., loc. cit.; O'Keefe et al., Biochem.
Biophys. Acta 1443 (1998), 113-127). Initial analysis of PSMA
demonstrated widespread expression within the cells of the
prostatic secretory epithelium. Immunohistochemical staining
demonstrated that PSMA was absent to moderately expressed in
hyperplastic and benign tissues, while malignant tissues stained
with the greatest intensity (Horoszewicz et al., loc. cit.).
Subsequent investigations have recapitulated these results and
evinced PSMA expression as a universal feature in practically every
prostatic tissue examined to date. These reports further
demonstrate that expression of PSMA increases precipitously
proportional to tumor aggressiveness (Burger et al., Int. J. Cancer
100 (2002), 228-237; Chang et al., Cancer Res. 59 (1999), 3192-98;
Chang et al., Urology 57 (2001), 1179-83), Kawakami and Nakayama,
Cancer Res. 57 (1997), 2321-24; Liu et al., Cancer Res. 57 (1997),
3629-34; Lopes et al., Cancer Res. 50 (1990), 6423-29; Silver et
al., Clin. Cancer Res. 9 (2003), 6357-62; Sweat et al., Urology 52
(1998), 637-40; Troyer et al., Int. J. Cancer 62 (1995), 552-558;
Wright et al., Urology 48 (1996), 326-334). Consistent with the
correlation between PSMA expression and tumor stage, increased
levels of PSMA are associated with androgen-independent prostate
cancer (PCa). Analysis of tissue samples from patients with
prostate cancer has demonstrated elevated PSMA levels after
physical castration or androgen-deprivation therapy. Unlike
expression of prostate specific antigen, which is downregulated
after androgen ablation, PSMA expression is significantly increased
in both primary and metastatic tumor specimens (Kawakami et al.,
Wright et al., loc. cit.). Consistent with the elevated expression
in androgen-independent tumors, PSMA transcription is also known to
be downregulated by steroids, and administration of testosterone
mediates a dramatic reduction in PSMA protein and mRNA levels
(Israeli et al., Cancer Res. 54 (1994), 1807-11; Wright et al.,
loc. cit.). PSMA is also highly expressed in secondary prostatic
tumors and occult metastatic disease. Immunohistochemical analysis
has revealed relatively intense and homogeneous expression of PSMA
within metastatic lesions localized to lymph nodes, bone, soft
tissue, and lungs compared with benign prostatic tissues (Chang et
al. (2001), loc. cit.; Murphy et al., Cancer 78 (1996), 809-818;
Sweat et al., loc. cit.). Some reports have also indicated limited
PSMA expression in extraprostatic tissues, including a subset of
renal proximal tubules, some cells of the intestinal brush-border
membrane, and rare cells in the colonic crypts (Chang et al.
(1999), Horoszewicz et al., Israeli et al. (1994), Lopes et al.,
Troyer et al., loc. cit.). However, the levels of PSMA in these
tissues are generally two to three orders of magnitude less than
those observed in the prostate (Sokoloff et al., Prostate 43
(2000), 150-157). PSMA is also expressed in the tumor-associated
neovasculature of most solid cancers examined yet is absent in the
normal vascular endothelium (Chang et al. (1999), Liu et al.,
Silver et al., loc. cit.). Although the significance of PSMA
expression within the vasculature is unknown, the specificity for
tumor-associated endothelium makes PSMA a potential target for the
treatment of many forms of malignancy.
[0011] Though there has been put much effort in identifying novel
targets for therapeutic approaches for cancer, cancer is yet one of
the most frequently diagnosed diseases. In light of this, there is
still need for effective treatments for cancer.
[0012] The present invention provides for a bispecific single chain
antibody molecule comprising a first binding domain capable of
binding to an epitope of human and non-chimpanzee primate
CD3.epsilon. (epsilon) chain, wherein the epitope is part of an
amino acid sequence comprised in the group consisting of SEQ ID
NOs. 2, 4, 6, and 8; and a second binding domain capable of binding
to prostate-specific membrane antigen (PSMA).
[0013] Though T cell-engaging bispecific single chain antibodies
described in the art have great therapeutic potential for the
treatment of malignant diseases, most of these bispecific molecules
are limited in that they are species specific and recognize only
human antigen, and--due to genetic similarity--likely the
chimpanzee counterpart. The advantage of the present invention is
the provision of a bispecific single chain antibody comprising a
binding domain exhibiting cross-species specificity to human and
non-chimpanzee primate of the CD3 epsilon chain.
[0014] In the present invention, an N-terminal 1-27 amino acid
residue polypeptide fragment of the extracellular domain of CD3
epsilon was surprisingly identified which--in contrast to all other
known epitopes of CD3 epsilon described in the art--maintains its
three-dimensional structural integrity when taken out of its native
environment in the CD3 complex (and optionally fused to a
heterologous amino acid sequence such as EpCAM or an immunoglobulin
Fc part). The present invention, therefore, provides for a
bispecific single chain antibody molecule comprising a first
binding domain capable of binding to an epitope of an N-terminal
1-27 amino acid residue polypeptide fragment of the extracellular
domain of CD3 epsilon (which CD3 epsilon is, for example, taken out
of its native environment and/or comprised by (presented on the
surface of) a T-cell) of human and at least one non-chimpanzee
primate CD3 epsilon chain, wherein the epitope is part of an amino
acid sequence comprised in the group consisting of SEQ ID NOs. 2,
4, 6, and 8; and a second binding domain capable of binding to
prostate-specific membrane antigen (PSMA). Preferred non-chimpanzee
primates are mentioned herein elsewhere. At least one (or a
selection thereof or all) primate(s) selected from Callithrix
jacchus; Saguinus oedipus, Saimiri sciureus, and Macaca
fascicularis (either SEQ ID 1047 or 1048 or both), is (are)
particularily preferred. Macaca mulatta, also known as Rhesus
Monkey is also envisaged as another preferred primate. It is thus
envisaged that antibodies of the invention bind to (are capable of
binding to) the context independent epitope of an N-terminal 1-27
amino acid residue polypeptide fragment of the extracellular domain
of CD3 epsilon of human and Callithrix jacchus, Saguinus oedipus,
Saimiri sciureus, and Macaca fascicularis (either SEQ ID 1047 or
1048 or both), and optionally also to Macaca mulatta. A bispecific
single chain antibody molecule comprising a first binding domain as
defined herein can be obtained (is obtainable by) or can be
manufactured in accordance with the protocol set out in the
appended Examples (in particular Example 2). To this end, it is
envisaged to (a) immunize mice with an N-terminal 1-27 amino acid
residue polypeptide fragment of the extracellular domain of CD3
epsilon of human and/or Saimiri sciureus; (b) generation of an
immune murine antibody scFv library; (c) identification of CD3
epsilon specific binders by testing the capability to bind to at
least SEQ ID NOs. 2, 4, 6, and 8.
[0015] The context-independence of the CD3 epitope provided in this
invention corresponds to the first 27 N-terminal amino acids of CD3
epsilon or functional fragments of this 27 amino acid stretch. The
phrase "context-independent," as used herein in relation to the CD3
epitope means that binding of the herein described inventive
binding molecules/antibody molecules does not lead to a change or
modification of the conformation, sequence, or structure
surrounding the antigenic determinant or epitope. In contrast, the
CD3 epitope recognized by a conventional CD3 binding molecule (e.g.
as disclosed in WO 99/54440 or WO 04/106380) is localized on the
CD3 epsilon chain C-terminally to the N-terminal 1-27 amino acids
of the context-independent epitope, where it only takes the correct
conformation if it is embedded within the rest of the epsilon chain
and held in the right sterical position by heterodimerization of
the epsilon chain with either the CD3 gamma or delta chain.
[0016] Anti-CD3 binding domains as part of a PSMA.times.CD3
bispecific single chain molecule as provided herein and generated
(and directed) against a context-independent CD3 epitope provide
for a surprising clinical improvement with regard to T cell
redistribution and, thus, a more favourable safety profile. Without
being bound by theory, since the CD3 epitope is
context-independent, forming an autonomous selfsufficient subdomain
without much influence on the rest of the CD3 complex, the CD3
binding domain of the PSMA.times.CD3 bispecific single chain
molecule provided herein induces less allosteric changes in CD3
conformation than the conventional CD3 binding molecules (like
molecules provided in WO 99/54440 or WO 04/106380), which recognize
context-dependent CD3 epitopes.
[0017] The context-independence of the CD3 epitope which is
recognized by the CD3 binding domain of the PSMA.times.CD3
bispecific single chain antibody of the invention is associated
with less or no T cell redistribution (T cell redistribution
equates with an initial episode of drop and subsequent recovery of
absolute T cell counts) during the starting phase of treatment with
said PSMA.times.CD3 bispecific single chain antibody of the
invention. This results in a better safety profile of the
PSMA.times.CD3 bispecific single chain antibody of the invention
compared to conventional CD3 binding molecules known in the art,
which recognize context-dependent CD3 epitopes. Particularly,
because T cell redistribution during the starting phase of
treatment with CD3 binding molecules is a major risk factor for
adverse events, like CNS adverse events, the PSMA.times.CD3
bispecific single chain antibody of the invention by recognizing a
context-independent rather than a context-dependent CD3 epitope has
a substantial safety advantage over the CD3 binding molecules known
in the art. Patients with such CNS adverse events related to T cell
redistribution during the starting phase of treatment with
conventional CD3 binding molecules usually suffer from confusion
and disorientation, in some cases also from urinary incontinence.
Confusion is a change in mental status in which the patient is not
able to think with his or her usual level of clarity. The patient
usually has difficulties to concentrate and thinking is not only
blurred and unclear but often significantly slowed down. Patients
with CNS adverse events related to T cell redistribution during the
starting phase of treatment with conventional CD3 binding molecules
may also suffer from loss of memory. Frequently, the confusion
leads to the loss of ability to recognize people, places, time or
the date. Feelings of disorientation are common in confusion, and
the decision-making ability is impaired. CNS adverse events related
to T cell redistribution during the starting phase of treatment
with conventional CD3 binding molecules may further comprise
blurred speech and/or word finding difficulties. This disorder may
impair both, the expression and understanding of language as well
as reading and writing. Besides urinary incontinence, vertigo and
dizziness may also accompany CNS adverse events related to T cell
redistribution during the starting phase of treatment with
conventional CD3 binding molecules in some patients.
[0018] The maintenance of the three-dimensional structure within
the mentioned 27 amino acid N-terminal polypeptide fragment of CD3
epsilon can be used for the generation of, preferably human,
binding domains which are capable of binding to the N-terminal CD3
epsilon polypeptide fragment in vitro and to the native (CD3
epsilon subunit of the) CD3 complex on T cells in vivo with the
same binding affinity. These data strongly indicate that the
N-terminal fragment as described herein forms a tertiary
conformation, which is similar to its structure normally existing
in vivo. A very sensitive test for the importance of the structural
integrity of the amino acids 1-27 of the N-terminal polypeptide
fragment of CD3 epsilon was performed. Individual amino acids of
amino acids 1-27 of the N-terminal polypeptide fragment of CD3
epsilon were changed to alanine (alanine scanning) to test the
sensitivity of the amino acids 1-27 of the N-terminal polypeptide
fragment of CD3 epsilon for minor disruptions. The CD3 specific
binding domains as part of the PSMA.times.CD3 bispecific single
chain antibody of the invention were used to test for binding to
the alanine-mutants of amino acids 1-27 of the N-terminal
polypeptide fragment of CD3 epsilon (see appended Example 5).
Individual exchanges of the first five amino acid residues at the
very N-terminal end of the fragment and two of the amino acids at
positions 23 and 25 of the amino acids 1-27 of the N-terminal
polypeptide fragment of CD3 epsilon were critical for binding of
the antibody molecules. The substitution of amino acid residues in
the region of position 1-5 comprising the residues Q (Glutamine at
position 1), D (Aspartic acid at position 2), G (Glycine at
position 3), N (Asparagine at position 4), and E (Glutamic acid at
position 5) to Alanine abolished binding of the, preferably human,
PSMA.times.CD3 bispecific single chain antibody of the invention to
said fragment. While, for at least some of the, preferably human,
PSMA.times.CD3 bispecific single chain antibody of the invention,
two amino acid residues at the C-terminus of the mentioned fragment
T (Threonine at position 23) and I (Isoleucine at position 25)
reduced the binding energy to the, preferably human, PSMA.times.CD3
bispecific single chain antibody of the invention.
[0019] Unexpectedly, it has been found that the thus isolated,
preferably human, PSMA.times.CD3 bispecific single chain antibody
of the invention not only recognizes the human N-terminal fragment
of CD3 epsilon, but also the corresponding homologous fragments of
CD3 epsilon of various primates, including New-World Monkeys
(Marmoset, Callithrix jacchus; Saguinus oedipus; Saimiri sciureus)
and Old-World Monkeys (Macaca fascicularis, also known as
Cynomolgus Monkey; or Macaca mulatta, also known as Rhesus Monkey).
Thus, multi-primate specificity of the PSMA.times.CD3 bispecific
single chain antibody of the invention was detected. The following
sequence analyses confirmed that human and primates share a highly
homologous sequence stretch at the N-terminus of the extracellular
domain of CD3 epsilon.
[0020] The amino acid sequence of the aformentioned N-terminal
fragments of CD3 epsilon are depicted in SEQ ID No. 2 (human), SEQ
ID No. 4 (Callithrix jacchus); SEQ ID No. 6 (Saguinus oedipus); SEQ
ID No. 8 (Saimiri sciureus); SEQ ID No. 1047
QDGNEEMGSITQTPYQVSISGTTILTC or SEQ ID No. 1048
QDGNEEMGSITQTPYQVSISGTTVILT (Macaca fascicularis, also known as
Cynomolgus Monkey), and SEQ ID No. 1049 QDGNEEMGSITQTPYHVSISGTTVILT
(Macaca mulatta, also known as Rhesus Monkey).
[0021] The second binding domain of the PSMA.times.CD3 bispecific
single chain antibody of the invention binds to the
prostate-specific membrane antigen (PSMA). Preferably, the second
binding domain of the PSMA.times.CD3 bispecific single chain
antibody binds to the human PSMA or a non-chimpanzee primate PSMA;
more preferred it binds to the human PSMA and a non-chimpanzee
primate PSMA and therefore is cross-species specific; even more
preferred to the human PSMA and the macaque PSMA (and therefore is
cross-species specific as well). Particularly preferred, the
macaque PSMA is the Cynomolgus monkey PSMA and/or the Rhesus monkey
PSMA. However, it is not excluded from the scope of the present
invention, that the second binding domain may also bind to PSMA
homologs of other species, such as to the PSMA homolog in
rodents.
[0022] Prostate cancer is the second most cancer in men. For 2008,
it is estimated that 186,320 men will be newly diagnosed with
prostate cancer in the United States and about 28,660 men will die
from the disease. Prostate cancer risk is strongly related to age:
very few cases are registered in men under 50 and three-quarters of
cases occur in men over 65 years. The largest number of cases is
diagnosed in those aged 70-74. Currently, the growth rate of the
older population is significantly higher than that of the total
population. By 2025-2030, projections indicate that the population
over 60 will be growing 3.5 times as rapidly as the total
population. The proportion of older persons is projected to more
than double worldwide over the next half century, which means that
a further increase in incidence of diagnosed prostate cancer has to
be expected for the future. The highly restricted expression of
PSMA and its upregulation in advanced stages and metastatic disease
of prostate cancer as well as its role as neoantigen on tumor
vasculature of many different types of other solid tumors qualifies
PSMA as attractive target antigen for antibody-based cancer
therapy. As shown in the following examples, the PSMA.times.CD3
bispecific single chain antibody of the invention provides an
advantageous tool in order to kill PSMA-expressing human cancer
cells, as exemplified by the human prostate cancer cell line LNCaP.
In addition, the cytotoxic activity of the PSMA.times.CD3
bispecific single chain antibody of the invention is higher than
the cytotoxic activity of antibodies described in the art. Since
preferably both the CD3 and the PSMA binding domain of the
PSMA.times.CD3 bispecific single chain antibody of the invention
are cross-species specific, i.e. reactive with the human and
non-chimpanzee primates antigens, it can be used for preclinical
evaluation of safety, activity and/or pharmacokinetic profile of
these binding domains in primates and--in the identical form--as
drug in humans.
[0023] Advantageously, the present invention provides also
PSMA.times.CD3 bispecific single chain antibodies comprising a
second binding domain which binds both to the human PSMA and to the
macaque PSMA homolog, i.e. the homolog of a non-chimpanzee primate.
In a preferred embodiment, the bispecific single chain antibody
thus comprises a second binding domain exhibiting cross-species
specificity to the human and a non-chimpanzee primate PSMA. In this
case, the identical bispecific single chain antibody molecule can
be used both for preclinical evaluation of safety, activity and/or
pharmacokinetic profile of these binding domains in primates and as
drug in humans. Put in other words, the same molecule can be used
in preclinical animal studies as well as in clinical studies in
humans. This leads to highly comparable results and a
much-increased predictive power of the animal studies compared to
species-specific surrogate molecules. Since both the CD3 and the
PSMA binding domain of the PSMA.times.CD3 bispecific single chain
antibody of the invention are cross-species specific, i.e. reactive
with the human and non-chimpanzee primates' antigens, it can be
used both for preclinical evaluation of safety, activity and/or
pharmacokinetic profile of these binding domains in primates
and--in the identical form--as drug in humans. It will be
understood that in a preferred embodiment, the cross-species
specificity of the first and second binding domain of the
antibodies of the invention is identical.
[0024] It has been found in the present invention that it is
possible to generate a, preferably human, PSMA.times.CD3 bispecific
single chain antibody wherein the identical molecule can be used in
preclinical animal testing, as well as clinical studies and even in
therapy in human. This is due to the unexpected identification of
the, preferably human, PSMA.times.CD3 bispecific single chain
antibody, which, in addition to binding to human CD3 epsilon and
PSMA, respectively, (and due to genetic similarity likely to the
chimpanzee counterpart), also binds to the homologs of said
antigens of non-chimpanzee primates, including New-World Monkeys
and Old-World Monkeys. As shown in the following Examples, said
preferably human, PSMA.times.CD3 bispecific single chain antibody
of the invention can be used as therapeutic agent against various
diseases, including, but not limited, to cancer. The PSMA.times.CD3
bispecific single chain antibody is particularly advantageous for
the therapy of cancer, preferably solid tumors, more preferably
carcinomas and prostate cancer. In view of the above, the need to
construct a surrogate PSMA.times.CD3 bispecific single chain
antibody for testing in a phylogenetic distant (from humans)
species disappears. As a result, the identical molecule can be used
in animal preclinical testing as is intended to be administered to
humans in clinical testing as well as following market approval and
therapeutic drug administration. The ability to use the same
molecule for preclinical animal testing as in later administration
to humans virtually eliminates, or at least greatly reduces, the
danger that the data obtained in preclinical animal testing have
limited applicability to the human case. In short, obtaining
preclinical safety data in animals using the same molecule as will
actually be administered to humans does much to ensure the
applicability of the data to a human-relevant scenario. In
contrast, in conventional approaches using surrogate molecules,
said surrogate molecules have to be molecularly adapted to the
animal test system used for preclinical safety assessment. Thus,
the molecule to be used in human therapy in fact differs in
sequence and also likely in structure from the surrogate molecule
used in preclinical testing in pharmacokinetic parameters and/or
biological activity, with the consequence that data obtained in
preclinical animal testing have limited
applicability/transferability to the human case. The use of
surrogate molecules requires the construction, production,
purification and characterization of a completely new construct.
This leads to additional development costs and time necessary to
obtain that molecule. In sum, surrogates have to be developed
separately in addition to the actual drug to be used in human
therapy, so that two lines of development for two molecules have to
be carried out. Therefore, a major advantage of the, preferably
human, PSMA.times.CD3 bispecific single chain antibody of the
invention exhibiting cross-species specificity described herein is
that the identical molecule can be used for therapeutic agents in
humans and in preclinical animal testing.
[0025] It is preferred that at least one of said first or second
binding domains of the bispecific single chain antibody of the
invention is CDR-grafted, humanized or human, as set forth in more
detail below. Preferably, both the first and second binding domains
of the bispecific single chain antibody of the invention are
CDR-grafted, humanized or human. For the preferably human,
PSMA.times.CD3 bispecific single chain antibody of the invention,
the generation of an immune reaction against said binding molecule
is excluded to the maximum possible extent upon administration of
the molecule to human patients.
[0026] Another major advantage of the, preferably human,
PSMA.times.CD3 bispecific single chain antibody of the invention is
its applicability for preclinical testing in various primates. The
behavior of a drug candidate in animals should ideally be
indicative of the expected behavior of this drug candidate upon
administration to humans. As a result, the data obtained from such
preclinical testing should therefore generally have a highly
predictive power for the human case. However, as learned from the
tragic outcome of the recent Phase I clinical trial on TGN1412 (a
CD28 monoclonal antibody), a drug candidate may act differently in
a primate species than in humans: Whereas in preclinical testing of
said antibody no or only limited adverse effects have been observed
in animal studies performed with cynomolgus monkeys, six human
patients developed multiple organ failure upon administration of
said antibody (Lancet 368 (2006), 2206-7). The results of these
dramatic, non-desired negative events suggest that it may not be
sufficient to limit preclinical testing to only one (non-chimpanzee
primate) species. The fact that the PSMA.times.CD3 bispecific
single chain antibody of the invention binds to a series of
New-World and Old-World Monkeys may help to overcome the problems
faced in the case mentioned above. Accordingly, the present
invention provides means and methods for minimizing species
differences in effects when drugs for human therapy are being
developed and tested.
[0027] With the, preferably human, cross-species specific
PSMA.times.CD3 bispecific single chain antibody of the invention it
is also no longer necessary to adapt the test animal to the drug
candidate intended for administration to humans, such as e.g. the
creation of transgenic animals. The, preferably human,
PSMA.times.CD3 bispecific single chain antibody of the invention
exhibiting cross-species specificity according to the uses and the
methods of invention can be directly used for preclinical testing
in non-chimpanzee primates, without any genetic manipulation of the
animals. As well known to those skilled in the art, approaches in
which the test animal is adapted to the drug candidate always bear
the risk that the results obtained in the preclinical safety
testing are less representative and predictive for humans due to
the modification of the animal. For example, in transgenic animals,
the proteins encoded by the transgenes are often highly
over-expressed. Thus, data obtained for the biological activity of
an antibody against this protein antigen may be limited in their
predictive value for humans in which the protein is expressed at
much lower, more physiological levels.
[0028] A further advantage of the uses of the preferably human
PSMA.times.CD3 bispecific single chain antibody of the invention
exhibiting cross-species specificity is the fact that chimpanzees
as an endangered species are avoided for animal testing.
[0029] Chimpanzees are the closest relatives to humans and were
recently grouped into the family of hominids based on the genome
sequencing data (Wildman et al., PNAS 100 (2003), 7181). Therefore,
data obtained with chimpanzee is generally considered to be highly
predictive for humans. However, due to their status as endangered
species, the number of chimpanzees, which can be used for medical
experiments, is highly restricted. As stated above, maintenance of
chimpanzees for animal testing is therefore both costly and
ethically problematic. The uses of the, preferably human,
PSMA.times.CD3 bispecific single chain antibody of the invention
avoid both ethical objections and financial burden during
preclinical testing without prejudicing the quality, i.e.
applicability, of the animal testing data obtained. In light of
this, the uses of the, preferably human, PSMA.times.CD3 bispecific
single chain antibody of the invention provide for a reasonable
alternative for studies in chimpanzees.
[0030] A still further advantage of the, preferably human,
PSMA.times.CD3 bispecific single chain antibody of the invention is
the ability of extracting multiple blood samples when using it as
part of animal preclinical testing, for example in the course of
pharmacokinetic animal studies. Multiple blood extractions can be
much more readily obtained with a non-chimpanzee primate than with
lower animals, e.g. a mouse. The extraction of multiple blood
samples allows continuous testing of blood parameters for the
determination of the biological effects induced by the, preferably
human, PSMA.times.CD3 bispecific single chain antibody of the
invention. Furthermore, the extraction of multiple blood samples
enables the researcher to evaluate the pharmacokinetic profile of
the, preferably human, PSMA.times.CD3 bispecific single chain
antibody of the invention as defined herein. In addition, potential
side effects, which may be induced by said, preferably human,
PSMA.times.CD3 bispecific single chain antibody of the invention
reflected in blood parameters can be measured in different blood
samples extracted during the course of the administration of said
antibody. This allows the determination of the potential toxicity
profile of the, preferably human, PSMA.times.CD3 bispecific single
chain antibody of the invention as defined herein.
[0031] The advantages of the, preferably human, PSMA.times.CD3
bispecific single chain antibody of the invention as defined herein
exhibiting cross-species specificity may be briefly summarized as
follows:
[0032] First, the, preferably human, PSMA.times.CD3 bispecific
single chain antibody of the invention as defined herein used in
preclinical testing is the same as the one used in human therapy.
Thus, it is no longer necessary to develop two independent
molecules, which may differ in their pharmacokinetic properties and
biological activity. This is highly advantageous in that e.g. the
pharmacokinetic results are more directly transferable and
applicable to the human setting than e.g. in conventional surrogate
approaches.
[0033] Second, the uses of the, preferably human, PSMA.times.CD3
bispecific single chain antibody of the invention as defined herein
for the preparation of therapeutics in human is less cost- and
labor-intensive than surrogate approaches.
[0034] Third, the, preferably human, PSMA.times.CD3 bispecific
single chain antibody of the invention as defined herein can be
used for preclinical testing not only in one primate species, but
in a series of different primate species, thereby limiting the risk
of potential species differences between primates and human.
[0035] Fourth, chimpanzee as an endangered species for animal
testing can be avoided if desired.
[0036] Fifth, multiple blood samples can be extracted for extensive
pharmacokinetic studies. Sixth, due to the human origin of the,
preferably human, binding molecules according to a preferred
embodiment of the invention, the generation of an immune reaction
against said binding molecules is minimalized when administered to
human patients. Induction of an immune response with antibodies
specific for a drug candidate derived from a non-human species as
e.g. a mouse leading to the development of human-anti-mouse
antibodies (HAMAs) against therapeutic molecules of murine origin
is excluded.
[0037] Last but not least, the therapeutic use of the
PSMA.times.CD3 bispecific single chain antibody of the invention
provides a novel and inventive therapeutic approach for cancer,
preferably solid tumors, more preferably carcinomas and prostate
cancer. As shown in the following examples, the PSMA.times.CD3
bispecific single chain antibody of the invention provides an
advantageous tool in order to kill PSMA-expressing human prostate
cancer cells. Moreover, the cytotoxic activity of the
PSMA.times.CD3 bispecific single chain antibody of the invention is
higher than the activity of antibodies described in the art.
[0038] As noted herein above, the present invention provides
polypeptides, i.e. bispecific single chain antibodies, comprising a
first binding domain capable of binding to an epitope of human and
non-chimpanzee primate CD3.epsilon. chain and a second binding
domain capable of binding to PSMA. The second binding domain
preferably binds to human PSMA and a non-chimpanzee primate PSMA.
The advantage of bispecific single chain antibody molecules as drug
candidates fulfilling the requirements of the preferred bispecific
single chain antibody of the invention is the use of such molecules
in preclinical animal testing as well as in clinical studies and
even for therapy in human. In a preferred embodiment of the
cross-species specific bispecific single chain antibodies of the
invention the second binding domain binding to PSMA is human. In a
cross-species specific bispecific molecule according to the
invention the binding domain binding to an epitope of human and
non-chimpanzee primate CD3 epsilon chain is located in the order
VH-VL or VL-VH at the N-terminus or the C-terminus of the
bispecific molecule. Examples for cross-species specific bispecific
molecules according to the invention in different arrangements of
the VH- and the VL-chain in the first and the second binding domain
are described in the appended examples.
[0039] As used herein, a "bispecific single chain antibody" denotes
a single polypeptide chain comprising two binding domains. Each
binding domain comprises one variable region from an antibody heavy
chain ("VH region"), wherein the VH region of the first binding
domain specifically binds to the CD3.epsilon. molecule, and the VH
region of the second binding domain specifically binds to PSMA. The
two binding domains are optionally linked to one another by a short
polypeptide spacer. A non-limiting example for a polypeptide spacer
is Gly-Gly-Gly-Gly-Ser (G-G-G-G-S) and repeats thereof. Each
binding domain may additionally comprise one variable region from
an antibody light chain ("VL region"), the VH region and VL region
within each of the first and second binding domains being linked to
one another via a polypeptide linker, for example of the type
disclosed and claimed in EP 623679 B1, but in any case long enough
to allow the VH region and VL region of the first binding domain
and the VH region and VL region of the second binding domain to
pair with one another such that, together, they are able to
specifically bind to the respective first and second binding
domains.
[0040] The term "protein" is well known in the art and describes
biological compounds. Proteins comprise one or more amino acid
chains (polypeptides), whereby the amino acids are bound among one
another via a peptide bond. The term "polypeptide" as used herein
describes a group of molecules, which consists of more than 30
amino acids. In accordance with the invention, the group of
polypeptides comprises "proteins" as long as the proteins consist
of a single polypeptide chain. Also in line with the definition the
term "polypeptide" describes fragments of proteins as long as these
fragments consist of more than 30 amino acids. Polypeptides may
further form multimers such as dimers, trimers and higher
oligomers, i.e. consisting of more than one polypeptide molecule.
Polypeptide molecules forming such dimers, trimers etc. may be
identical or non-identical. The corresponding higher order
structures of such multimers are, consequently, termed homo- or
heterodimers, homo- or heterotrimers etc. An example for a
hereteromultimer is an antibody molecule, which, in its naturally
occurring form, consists of two identical light polypeptide chains
and two identical heavy polypeptide chains. The terms "polypeptide"
and "protein" also refer to naturally modified
polypeptides/proteins wherein the modification is effected e.g. by
post-translational modifications like glycosylation, acetylation,
phosphorylation and the like. Such modifications are well known in
the art.
[0041] The term "binding domain" characterizes in connection with
the present invention a domain of a polypeptide which specifically
binds to/interacts with a given target structure/antigen/epitope.
Thus, the binding domain is an "antigen-interaction-site". The term
"antigen-interaction-site" defines, in accordance with the present
invention, a motif of a polypeptide, which is able to specifically
interact with a specific antigen or a specific group of antigens,
e.g. the identical antigen in different species. Said
binding/interaction is also understood to define a "specific
recognition". The term "specifically recognizing" means in
accordance with this invention that the antibody molecule is
capable of specifically interacting with and/or binding to at least
two, preferably at least three, more preferably at least four amino
acids of an antigen, e.g. the human CD3 antigen as defined herein.
Such binding may be exemplified by the specificity of a
"lock-and-key-principle". Thus, specific motifs in the amino acid
sequence of the binding domain and the antigen bind to each other
as a result of their primary, secondary or tertiary structure as
well as the result of secondary modifications of said structure.
The specific interaction of the antigen-interaction-site with its
specific antigen may result as well in a simple binding of said
site to the antigen. Moreover, the specific interaction of the
binding domain/antigen-interaction-site with its specific antigen
may alternatively result in the initiation of a signal, e.g. due to
the induction of a change of the conformation of the antigen, an
oligomerization of the antigen, etc. A preferred example of a
binding domain in line with the present invention is an antibody.
The binding domain may be a monoclonal or polyclonal antibody or
derived from a monoclonal or polyclonal antibody.
[0042] The term "antibody" comprises derivatives or functional
fragments thereof which still retain the binding specificity.
Techniques for the production of antibodies are well known in the
art and described, e.g. in Harlow and Lane "Antibodies, A
Laboratory Manual", Cold Spring Harbor Laboratory Press, 1988 and
Harlow and Lane "Using Antibodies: A Laboratory Manual" Cold Spring
Harbor Laboratory Press, 1999. The term "antibody" also comprises
immunoglobulins (Ig's) of different classes (i.e. IgA, IgG, IgM,
IgD and IgE) and subclasses (such as IgG1, IgG2 etc.).
[0043] The definition of the term "antibody" also includes
embodiments such as chimeric, single chain and humanized
antibodies, as well as antibody fragments, like, inter alia, Fab
fragments. Antibody fragments or derivatives further comprise
F(ab').sub.2, Fv, scFv fragments or single domain antibodies,
single variable domain antibodies or immunoglobulin single variable
domain comprising merely one variable domain, which might be VH or
VL, that specifically bind to an antigen or epitope independently
of other V regions or domains; see, for example, Harlow and Lane
(1988) and (1999), loc. cit. Such immunoglobulin single variable
domain encompasses not only an isolated antibody single variable
domain polypeptide, but also larger polypeptides that comprise one
or more monomers of an antibody single variable domain polypeptide
sequence.
[0044] Various procedures are known in the art and may be used for
the production of such antibodies and/or fragments. Thus, the
(antibody) derivatives can also be produced by peptidomimetics.
Further, techniques described for the production of single chain
antibodies (see, inter alia, U.S. Pat. No. 4,946,778) can be
adapted to produce single chain antibodies specific for elected
polypeptide(s). Also, transgenic animals may be used to express
humanized or human antibodies specific for polypeptides and fusion
proteins of this invention. For the preparation of monoclonal
antibodies, any technique, providing antibodies produced by
continuous cell line cultures can be used. Examples for such
techniques include the hybridoma technique (Kohler and Milstein
Nature 256 (1975), 495-497), the trioma technique, the human B-cell
hybridoma technique (Kozbor, Immunology Today 4 (1983), 72) and the
EBV-hybridoma technique to produce human monoclonal antibodies
(Cole et al., Monoclonal Antibodies and Cancer Therapy, Alan R.
Liss, Inc. (1985), 77-96). Surface plasmon resonance as employed in
the BIAcore system can be used to increase the efficiency of phage
antibodies which bind to an epitope of a target polypeptide, such
as CD3 epsilon or PSMA (Schier, Human Antibodies Hybridomas 7
(1996), 97-105; Malmborg, J. Immunol. Methods 183 (1995), 7-13). It
is also envisaged in the context of this invention that the term
"antibody" comprises antibody constructs, which may be expressed in
a host as described herein below, e.g. antibody constructs which
may be transfected and/or transduced via, inter alia, viruses or
plasm id vectors.
[0045] The term "specific interaction" as used in accordance with
the present invention means that the binding domain does not or
does not significantly cross-react with polypeptides which have
similar structure as those bound by the binding domain, and which
might be expressed by the same cells as the polypeptide of
interest. Cross-reactivity of a panel of binding domains under
investigation may be tested, for example, by assessing binding of
said panel of binding domains under conventional conditions (see,
e.g., Harlow and Lane, Antibodies: A Laboratory Manual, Cold Spring
Harbor Laboratory Press, 1988 and Using Antibodies: A Laboratory
Manual, Cold Spring Harbor Laboratory Press, 1999). Examples for
the specific interaction of a binding domain with a specific
antigen comprise the specificity of a ligand for its receptor. Said
definition particularly comprises the interaction of ligands, which
induce a signal upon binding to its specific receptor. Examples for
said interaction, which is also particularly comprised by said
definition, is the interaction of an antigenic determinant
(epitope) with the binding domain (antigenic binding site) of an
antibody.
[0046] The term "cross-species specificity" or "interspecies
specificity" as used herein means binding of a binding domain
described herein to the same target molecule in humans and
non-chimpanzee primates. Thus, "cross-species specificity" or
"interspecies specificity" is to be understood as an interspecies
reactivity to the same molecule "X" expressed in different species,
but not to a molecule other than "X". Cross-species specificity of
a monoclonal antibody recognizing e.g. human CD3 epsilon, to a
non-chimpanzee primate CD3 epsilon, e.g. macaque CD3 epsilon, can
be determined, for instance, by FACS analysis. The FACS analysis is
carried out in a way that the respective monoclonal antibody is
tested for binding to human and non-chimpanzee primate cells, e.g.
macaque cells, expressing said human and non-chimpanzee primate CD3
epsilon antigens, respectively. An appropriate assay is shown in
the following examples. The above-mentioned subject matter applies
mutatis mutandis for the PSMA antigen: Cross-species specificity of
a monoclonal antibody recognizing e.g. human PSMA, to a
non-chimpanzee primate PSMA, e.g. macaque PSMA, can be determined,
for instance, by FACS analysis. The FACS analysis is carried out in
a way that the respective monoclonal antibody is tested for binding
to human and non-chimpanzee primate cells, e.g. macaque cells,
expressing said human and non-chimpanzee primate PSMA antigens,
respectively.
[0047] As used herein, CD3 epsilon denotes a molecule expressed as
part of the T cell receptor and has the meaning as typically
ascribed to it in the prior art. In human, it encompasses in
individual or independently combined form all known CD3 subunits,
for example CD3 epsilon, CD3 delta, CD3 gamma, CD3 zeta, CD3 alpha
and CD3 beta. The non-chimpanzee primate, non-human CD3 antigens as
referred to herein are, for example, Macaca fascicularis CD3 and
Macaca mulatta CD3. In Macaca fascicularis, it encompasses CD3
epsilon FN-18 negative and CD3 epsilon FN-18 positive, CD3 gamma
and CD3 delta. In Macaca mulatta, it encompasses CD3 epsilon, CD3
gamma and CD3 delta. Preferably, said CD3 as used herein is CD3
epsilon.
[0048] The human CD3 epsilon is indicated in GENBANK.RTM. Accession
No. NM_000733 and comprises SEQ ID NO. 1. The human CD3 gamma is
indicated in GENBANK.RTM. Accession NO. NM_000073. The human CD3
delta is indicated in GENBANK.RTM. Accession No. NM_000732.
[0049] The CD3 epsilon "FN-18 negative" of Macaca fascicularis
(i.e. CD3 epsilon not recognized by monoclonal antibody FN-18 due
to a polymorphism as set forth above) is indicated in GENBANK.RTM.
Accession No. AB073994.
[0050] The CD3 epsilon "FN-18 positive" of Macaca fascicularis
(i.e. CD3 epsilon recognized by monoclonal antibody FN-18) is
indicated in GENBANK.RTM. Accession No. AB073993. The CD3 gamma of
Macaca fascicularis is indicated in GENBANK.RTM. Accession No.
AB073992. The CD3 delta of Macaca fascicularis is indicated in
GENBANK.RTM. Accession No. AB073991.
[0051] The nucleic acid sequences and amino acid sequences of the
respective CD3 epsilon, gamma and delta homologs of Macaca mulatta
can be identified and isolated by recombinant techniques described
in the art (Sambrook et al. Molecular Cloning: A Laboratory Manual;
Cold Spring Harbor Laboratory Press, 3.sup.rd edition 2001). This
applies mutatis mutandis to the CD3 epsilon, gamma and delta
homologs of other non-chimpanzee primates as defined herein. The
identification of the amino acid sequence of Callithrix jacchus,
Saimiri sciureus and Saguinus oedipus is described in the appended
examples. The amino acid sequence of the extracellular domain of
the CD3 epsilon of Callithrix jacchus is depicted in SEQ ID NO: 3,
the one of Saguinus oedipus is depicted in SEQ ID NO: 5 and the one
of Saimiri sciureus is depicted in SEQ ID NO: 7.
[0052] The human PSMA is indicated in GENBANK.RTM. Accession No.
`AY101595`. The cloning of the PSMA homolog of macaque is
demonstrated in the following examples, the corresponding cDNA and
amino acid sequences are shown in SEQ ID NOs. 385 and 386,
respectively.
[0053] In line with the above, the term "epitope" defines an
antigenic determinant, which is specifically bound/identified by a
binding domain as defined herein. The binding domain may
specifically bind to/interact with conformational or continuous
epitopes, which are unique for the target structure, e.g. the human
and non-chimpanzee primate CD3 epsilon chain or the human and
non-chimpanzee primate PSMA. A conformational or discontinuous
epitope is characterized for polypeptide antigens by the presence
of two or more discrete amino acid residues which are separated in
the primary sequence, but come together on the surface of the
molecule when the polypeptide folds into the native protein/antigen
(Sela, (1969) Science 166, 1365 and Laver, (1990) Cell 61, 553-6).
The two or more discrete amino acid residues contributing to the
epitope are present on separate sections of one or more polypeptide
chain(s). These residues come together on the surface of the
molecule when the polypeptide chain(s) fold(s) into a
three-dimensional structure to constitute the epitope. In contrast,
a continuous or linear epitope consists of two or more discrete
amino acid residues, which are present in a single linear segment
of a polypeptide chain. Within the present invention, a
"context-dependent" CD3 epitope refers to the conformation of said
epitope. Such a context-dependent epitope, localized on the epsilon
chain of CD3, can only develop its correct conformation if it is
embedded within the rest of the epsilon chain and held in the right
position by heterodimerization of the epsilon chain with either CD3
gamma or delta chain. In contrast, a context-independent CD3
epitope as provided herein refers to an N-terminal 1-27 amino acid
residue polypeptide or a functional fragment thereof of CD3
epsilon. This N-terminal 1-27 amino acid residue polypeptide or a
functional fragment thereof maintains its three-dimensional
structural integrity and correct conformation when taken out of its
native environment in the CD3 complex. The context-independency of
the N-terminal 1-27 amino acid residue polypeptide or a functional
fragment thereof, which is part of the extracellular domain of CD3
epsilon, represents, thus, an epitope which is completely different
to the epitopes of CD3 epsilon described in connection with a
method for the preparation of human binding molecules in WO
2004/106380. Said method used solely expressed recombinant CD3
epsilon. The conformation of this solely expressed recombinant CD3
epsilon differed from that adopted in its natural form, that is,
the form in which the CD3 epsilon subunit of the TCR/CD3 complex
exists as part of a noncovalent complex with either the CD3 delta
or the CD3-gamma subunit of the TCR/CD3 complex. When such solely
expressed recombinant CD3 epsilon protein is used as an antigen for
selection of antibodies from an antibody library, antibodies
specific for this antigen are identified from the library although
such a library does not contain antibodies with specificity for
self-antigens/autoantigens. This is due to the fact that solely
expressed recombinant CD3 epsilon protein does not exist in vivo;
it is not an autoantigen. Consequently, subpopulations of B cells
expressing antibodies specific for this protein have not been
depleted in vivo; an antibody library constructed from such B cells
would contain genetic material for antibodies specific for solely
expressed recombinant CD3 epsilon protein.
[0054] However, since the context-independent N-terminal 1-27 amino
acid residue polypeptide or a functional fragment thereof is an
epitope, which folds in its native form, binding domains in line
with the present invention cannot be identified by methods based on
the approach described in WO 2004/106380. Therefore, it could be
verified in tests that binding molecules as disclosed in WO
2004/106380 are not capable of binding to the N-terminal 1-27 amino
acid residues of the CD3 epsilon chain. Hence, conventional
anti-CD3 binding molecules or anti-CD3 antibody molecules (e.g. as
disclosed in WO 99/54440) bind CD3 epsilon chain at a position
which is more C-terminally located than the context-independent
N-terminal 1-27 amino acid residue polypeptide or a functional
fragment provided herein. Prior art antibody molecules OKT3 and
UCHT-1 have also a specificity for the epsilon-subunit of the
TCR/CD3 complex between amino acid residues 35 to 85 and,
accordingly, the epitope of these antibodies is also more
C-terminally located. In addition, UCHT-1 binds to the CD3 epsilon
chain in a region between amino acid residues 43 to 77
(Tunnacliffe, Int. Immunol. 1 (1989), 546-50; Kjer-Nielsen, PNAS
101, (2004), 7675-7680; Salmeron, J. Immunol. 147 (1991), 3047-52).
Therefore, prior art anti-CD3 molecules do not bind to and are not
directed against the herein defined context-independent N-terminal
1-27 amino acid residue epitope (or a functional fragment thereof).
In particular, the state of the art fails to provide anti-CD3
molecules which specifically binds to the context-independent
N-terminal 1-27 amino acid residue epitope and which are
cross-species specific, i.e. bind to human and non-chimpanzee
primate CD3 epsilon.
[0055] For the generation of a, preferably human, binding domain
comprised in a bispecific single chain antibody molecule of the
invention, e.g. monoclonal antibodies binding to both the human and
non-chimpanzee primate CD3 epsilon (e.g. macaque CD3 epsilon) or
monoclonal antibodies binding to both the human and non-chimpanzee
primate PSMA can be used.
[0056] As used herein, "human" and "man" refers to the species Homo
sapiens. As far as the medical uses of the constructs described
herein are concerned, human patients are to be treated with the
same molecule.
[0057] It is preferred that at least one of said first or second
binding domains of the bispecific single chain antibody of the
invention is CDR-grafted, humanized or human. Preferably, both the
first and second binding domains of the bispecific single chain
antibody of the invention are CDR-grafted, humanized or human.
[0058] The term "human" antibody as used herein is to be understood
as meaning that the bispecific single chain antibody as defined
herein, comprises (an) amino acid sequence(s) contained in the
human germline antibody repertoire. For the purposes of definition
herein, said bispecific single chain antibody may therefore be
considered human if it consists of such (a) human germline amino
acid sequence(s), i.e. if the amino acid sequence(s) of the
bispecific single chain antibody in question is (are) identical to
(an) expressed human germline amino acid sequence(s). A bispecific
single chain antibody as defined herein may also be regarded as
human if it consists of (a) sequence(s) that deviate(s) from its
(their) closest human germline sequence(s) by no more than would be
expected due to the imprint of somatic hypermutation. Additionally,
the antibodies of many non-human mammals, for example rodents such
as mice and rats, comprise VH CDR3 amino acid sequences which one
may expect to exist in the expressed human antibody repertoire as
well. Any such sequence(s) of human or non-human origin which may
be expected to exist in the expressed human repertoire would also
be considered "human" for the purposes of the present
invention.
[0059] As used herein, the term "humanized", "humanization",
"human-like" or grammatically related variants thereof are used
interchangeably to refer to a bispecific single chain antibody
comprising in at least one of its binding domains at least one
complementarity determining region ("CDR") from a non-human
antibody or fragment thereof. Humanization approaches are described
for example in WO 91/09968 and U.S. Pat. No. 6,407,213. As
non-limiting examples, the term encompasses the case in which a
variable region of at least one binding domain comprises a single
CDR region, for example the third CDR region of the VH (CDRH3),
from another non-human animal, for example a rodent, as well as the
case in which a or both variable region/s comprise at each of their
respective first, second and third CDRs the CDRs from said
non-human animal. In the event that all CDRs of a binding domain of
the bispecific single chain antibody have been replaced by their
corresponding equivalents from, for example, a rodent, one
typically speaks of "CDR-grafting", and this term is to be
understood as being encompassed by the term "humanized" or
grammatically related variants thereof as used herein. The term
"humanized" or grammatically related variants thereof also
encompasses cases in which, in addition to replacement of one or
more CDR regions within a VH and/or VL of the first and/or second
binding domain further mutation/s (e.g. substitutions) of at least
one single amino acid residue/s within the framework ("FR") regions
between the CDRs has/have been effected such that the amino acids
at that/those positions correspond/s to the amino acid/s at
that/those position/s in the animal from which the CDR regions used
for replacement is/are derived. As is known in the art, such
individual mutations are often made in the framework regions
following CDR-grafting in order to restore the original binding
affinity of the non-human antibody used as a CDR-donor for its
target molecule. The term "humanized" may further encompass (an)
amino acid substitution(s) in the CDR regions from a non-human
animal to the amino acid(s) of a corresponding CDR region from a
human antibody, in addition to the amino acid substitutions in the
framework regions as described above.
[0060] As used herein, the term "homolog" or "homology" is to be
understood as follows: Homology among proteins and DNA is often
concluded on the basis of sequence similarity, especially in
bioinformatics. For example, in general, if two or more genes have
highly similar DNA sequences, it is likely that they are
homologous. But sequence similarity may arise from different
ancestors: short sequences may be similar by chance, and sequences
may be similar because both were selected to bind to a particular
protein, such as a transcription factor. Such sequences are similar
but not homologous. Sequence regions that are homologous are also
called conserved. This is not to be confused with conservation in
amino acid sequences in which the amino acid at a specific position
has changed but the physio-chemical properties of the amino acid
remain unchanged. Homologous sequences are of two types:
orthologous and paralogous. Homologous sequences are orthologous if
they were separated by a speciation event: when a species diverges
into two separate species, the divergent copies of a single gene in
the resulting species are said to be orthologous. Orthologs, or
orthologous genes, are genes in different species that are similar
to each other because they originated from a common ancestor. The
strongest evidence that two similar genes are orthologous is the
result of a phylogenetic analysis of the gene lineage. Genes that
are found within one clade are orthologs, descended from a common
ancestor. Orthologs often, but not always, have the same function.
Orthologous sequences provide useful information in taxonomic
classification studies of organisms. The pattern of genetic
divergence can be used to trace the relatedness of organisms. Two
organisms that are very closely related are likely to display very
similar DNA sequences between two orthologs. Conversely, an
organism that is further removed evolutionarily from another
organism is likely to display a greater divergence in the sequence
of the orthologs being studied. Homologous sequences are paralogous
if they were separated by a gene duplication event: if a gene in an
organism is duplicated to occupy two different positions in the
same genome, then the two copies are paralogous. A set of sequences
that are paralogous are called paralogs of each other. Paralogs
typically have the same or similar function, but sometimes do not:
due to lack of the original selective pressure upon one copy of the
duplicated gene, this copy is free to mutate and acquire new
functions. An example can be found in rodents such as rats and
mice. Rodents have a pair of paralogous insulin genes, although it
is unclear if any divergence in function has occurred. Paralogous
genes often belong to the same species, but this is not necessary:
for example, the hemoglobin gene of humans and the myoglobin gene
of chimpanzees are paralogs. This is a common problem in
bioinformatics: when genomes of different species have been
sequenced and homologous genes have been found, one can not
immediately conclude that these genes have the same or similar
function, as they could be paralogs whose function has
diverged.
[0061] As used herein, a "non-chimpanzee primate" or "non-chimp
primate" or grammatical variants thereof refers to any primate
animal (i.e. not human) other than chimpanzee, i.e. other than an
animal of belonging to the genus Pan, and including the species Pan
paniscus and Pan troglodytes, also known as Anthropopithecus
troglodytes or Simia satyrus. It will be understood, however, that
it is possible that the antibodies of the invention can also bind
with their first and/or second binding domain to the respective
epitopes/fragments etc. of said chimpanzees. The intention is
merely to avoid animal tests which are carried out with
chimpanzees, if desired. It is thus also envisaged that in another
embodiment the antibodies of the present invention also bind with
their first and/or second binding domain to the respective epitopes
of chimpanzees. A "primate", "primate species", "primates" or
grammatical variants thereof denote/s an order of eutherian mammals
divided into the two suborders of prosimians and anthropoids and
comprising apes, monkeys and lemurs. Specifically, "primates" as
used herein comprises the suborder Strepsirrhini (non-tarsier
prosimians), including the infraorder Lemuriformes (itself
including the superfamilies Cheirogaleoidea and Lemuroidea), the
infraorder Chiromyiformes (itself including the family
Daubentoniidae) and the infraorder Lorisiformes (itself including
the families Lorisidae and Galagidae). "Primates" as used herein
also comprises the suborder Haplorrhini, including the infraorder
Tarsiiformes (itself including the family Tarsiidae), the
infraorder Simiiformes (itself including the Platyrrhini, or
New-World monkeys, and the Catarrhini, including the
Cercopithecidea, or Old-World Monkeys).
[0062] The non-chimpanzee primate species may be understood within
the meaning of the invention to be a lemur, a tarsier, a gibbon, a
marmoset (belonging to New-World Monkeys of the family Cebidae) or
an Old-World Monkey (belonging to the superfamily
Cercopithecoidea).
[0063] As used herein, an "Old-World Monkey" comprises any monkey
falling in the superfamily Cercopithecoidea, itself subdivided into
the families: the Cercopithecinae, which are mainly African but
include the diverse genus of macaques which are Asian and North
African; and the Colobinae, which include most of the Asian genera
but also the African colobus monkeys.
[0064] Specifically, within the subfamily Cercopithecinae, an
advantageous non-chimpanzee primate may be from the Tribe
Cercopithecini, within the genus Allenopithecus (Allen's Swamp
Monkey, Allenopithecus nigroviridis); within the genus Miopithecus
(Angolan Talapoin, Miopithecus talapoin; Gabon Talapoin,
Miopithecus ogouensis); within the genus Erythrocebus (Patas
Monkey, Erythrocebus patas); within the genus Chlorocebus (Green
Monkey, Chlorocebus sabaceus; Grivet, Chlorocebus aethiops; Bale
Mountains Vervet, Chlorocebus djamdjamensis; Tantalus Monkey,
Chlorocebus tantalus; Vervet Monkey, Chlorocebus pygerythrus;
Malbrouck, Chlorocebus cynosuros); or within the genus
Cercopithecus (Dryas Monkey or Salongo Monkey, Cercopithecus dryas;
Diana Monkey, Cercopithecus diana; Roloway Monkey, Cercopithecus
roloway; Greater Spot-nosed Monkey, Cercopithecus nictitans; Blue
Monkey, Cercopithecus mitis; Silver Monkey, Cercopithecus doggetti;
Golden Monkey, Cercopithecus kandti; Sykes's Monkey, Cercopithecus
albogularis; Mona Monkey, Cercopithecus mona; Campbell's Mona
Monkey, Cercopithecus campbelli; Lowe's Mona Monkey, Cercopithecus
lowei; Crested Mona Monkey, Cercopithecus pogonias; Wolf's Mona
Monkey, Cercopithecus wolfi; Dent's Mona Monkey, Cercopithecus
denti; Lesser Spot-nosed Monkey, Cercopithecus petaurista;
White-throated Guenon, Cercopithecus erythrogaster; Sclater's
Guenon, Cercopithecus sclateri; Red-eared Guenon, Cercopithecus
erythrotis; Moustached Guenon, Cercopithecus cephus; Red-tailed
Monkey, Cercopithecus ascanius; L'Hoest's Monkey, Cercopithecus
lhoesti; Preuss's Monkey, Cercopithecus preussi; Sun-tailed Monkey,
Cercopithecus solatus; Hamlyn's Monkey or Owl-faced Monkey,
Cercopithecus hamlyni; De Brazza's Monkey, Cercopithecus
neglectus).
[0065] Alternatively, an advantageous non-chimpanzee primate, also
within the subfamily Cercopithecinae but within the Tribe
Papionini, may be from within the genus Macaca (Barbary Macaque,
Macaca sylvanus; Lion-tailed Macaque, Macaca silenus; Southern
Pig-tailed Macaque or Beruk, Macaca nemestrina; Northern Pig-tailed
Macaque, Macaca leonina; Pagai Island Macaque or Bokkoi, Macaca
pagensis; Siberut Macaque, Macaca siberu; Moor Macaque, Macaca
maura; Booted Macaque, Macaca ochreata; Tonkean Macaque, Macaca
tonkeana; Heck's Macaque, Macaca hecki; Gorontalo Macaque, Macaca
nigriscens; Celebes Crested Macaque or Black "Ape", Macaca nigra;
Cynomolgus monkey or Crab-eating Macaque or Long-tailed Macaque or
Kera, Macaca fascicularis; Stump-tailed Macaque or Bear Macaque,
Macaca arctoides; Rhesus Macaque, Macaca mulatta; Formosan Rock
Macaque, Macaca cyclopis; Japanese Macaque, Macaca fuscata; Toque
Macaque, Macaca sinica; Bonnet Macaque, Macaca radiata; Barbary
Macaque, Macaca sylvanmus; Assam Macaque, Macaca assamensis;
Tibetan Macaque or Milne-Edwards' Macaque, Macaca thibetana;
Arunachal Macaque or Munzala, Macaca munzala); within the genus
Lophocebus (Gray-cheeked Mangabey, Lophocebus albigena; Lophocebus
albigena albigena; Lophocebus albigena osmani; Lophocebus albigena
johnstoni; Black Crested Mangabey, Lophocebus aterrimus;
Opdenbosch's Mangabey, Lophocebus opdenboschi; Highland Mangabey,
Lophocebus kipunji); within the genus Papio (Hamadryas Baboon,
Papio hamadryas; Guinea Baboon, Papio papio; Olive Baboon, Papio
anubis; Yellow Baboon, Papio cynocephalus; Chacma Baboon, Papio
ursinus); within the genus Theropithecus (Gelada, Theropithecus
gelada); within the genus Cercocebus (Sooty Mangabey, Cercocebus
atys; Cercocebus atys atys; Cercocebus atys lunulatus; Collared
Mangabey, Cercocebus torquatus; Agile Mangabey, Cercocebus agilis;
Golden-bellied Mangabey, Cercocebus chrysogaster; Tana River
Mangabey, Cercocebus galeritus; Sanje Mangabey, Cercocebus sanjei);
or within the genus Mandrillus (Mandrill, Mandrillus sphinx; Drill,
Mandrillus leucophaeus).
[0066] Most preferred is Macaca fascicularis (also known as
Cynomolgus monkey and, therefore, in the Examples named
"Cynomolgus") and Macaca mulatta (rhesus monkey, named
"rhesus").
[0067] Within the subfamily Colobinae, an advantageous
non-chimpanzee primate may be from the African group, within the
genus Colobus (Black Colobus, Colobus satanas; Angola Colobus,
Colobus angolensis; King Colobus, Colobus polykomos; Ursine
Colobus, Colobus vellerosus; Mantled Guereza, Colobus guereza);
within the genus Piliocolobus (Western Red Colobus, Piliocolobus
badius; Piliocolobus badius badius; Piliocolobus badius temminckii;
Piliocolobus badius waldronae; Pennant's Colobus, Piliocolobus
pennantii; Piliocolobus pennantii pennantii; Piliocolobus pennantii
epieni; Piliocolobus pennantii bouvieri; Preuss's Red Colobus,
Piliocolobus preussi; Thollon's Red Colobus, Piliocolobus tholloni;
Central African Red Colobus, Piliocolobus foai; Piliocolobus foai
foai; Piliocolobus foai ellioti; Piliocolobus foai oustaleti;
Piliocolobus foai semlikiensis; Piliocolobus foai parmentierorum;
Ugandan Red Colobus, Piliocolobus tephrosceles; Uzyngwa Red
Colobus, Piliocolobus gordonorum; Zanzibar Red Colobus,
Piliocolobus kirkii; Tana River Red Colobus, Piliocolobus
rufomitratus); or within the genus Procolobus (Olive Colobus,
Procolobus verus).
[0068] Within the subfamily Colobinae, an advantageous
non-chimpanzee primate may alternatively be from the Langur (leaf
monkey) group, within the genus Semnopithecus (Nepal Gray Langur,
Semnopithecus schistaceus; Kashmir Gray Langur, Semnopithecus ajax;
Tarai Gray Langur, Semnopithecus hector, Northern Plains Gray
Langur, Semnopithecus entellus; Black-footed Gray Langur,
Semnopithecus hypoleucos; Southern Plains Gray Langur,
Semnopithecus dussumieri; Tufted Gray Langur, Semnopithecus priam);
within the T. vetulus group or the genus Trachypithecus
(Purple-faced Langur, Trachypithecus vetulus; Nilgiri Langur,
Trachypithecus johnii); within the T. cristatus group of the genus
Trachypithecus (Javan Lutung, Trachypithecus auratus; Silvery Leaf
Monkey or Silvery Lutung, Trachypithecus cristatus; Indochinese
Lutung, Trachypithecus germaini; Tenasserim Lutung, Trachypithecus
barbel); within the T. obscurus group of the genus Trachypithecus
(Dusky Leaf Monkey or Spectacled Leaf Monkey, Trachypithecus
obscurus; Phayre's Leaf Monkey, Trachypithecus phayrei); within the
T. pileatus group of the genus Trachypithecus (Capped Langur,
Trachypithecus pileatus; Shortridge's Langur, Trachypithecus
shortridgei; Gee's Golden Langur, Trachypithecus geei); within the
T. francoisi group of the genus Trachypithecus (Francois' Langur,
Trachypithecus francoisi; Hatinh Langur, Trachypithecus
hatinhensis; White-headed Langur, Trachypithecus poliocephalus;
Laotian Langur, Trachypithecus laotum; Delacour's Langur,
Trachypithecus delacouri; Indochinese Black Langur, Trachypithecus
ebenus); or within the genus Presbytis (Sumatran Surili, Presbytis
melalophos; Banded Surili, Presbytis femoralis; Sarawak Surili,
Presbytis chrysomelas; White-thighed Surili, Presbytis siamensis;
White-fronted Surili, Presbytis frontata; Javan Surili, Presbytis
comata; Thomas's Langur, Presbytis thomasi; Hose's Langur,
Presbytis hosei; Maroon Leaf Monkey, Presbytis rubicunda; Mentawai
Langur or Joja, Presbytis potenziani; Natuna Island Surili,
Presbytis natunae).
[0069] Within the subfamily Colobinae, an advantageous
non-chimpanzee primate may alternatively be from the Odd-Nosed
group, within the genus Pygathrix (Red-shanked Douc, Pygathrix
nemaeus; Black-shanked Douc, Pygathrix nigripes; Gray-shanked Douc,
Pygathrix cinerea); within the genus Rhinopithecus (Golden
Snub-nosed Monkey, Rhinopithecus roxellana; Black Snub-nosed
Monkey, Rhinopithecus bieti; Gray Snub-nosed Monkey, Rhinopithecus
brelichi; Tonkin Snub-nosed Langur, Rhinopithecus avunculus);
within the genus Nasalis (Proboscis Monkey, Nasalis larvatus); or
within the genus Simias (Pig-tailed Langur, Simias concolor).
[0070] As used herein, the term "marmoset" denotes any New-World
Monkeys of the genus Callithrix, for example belonging to the
Atlantic marmosets of subgenus Callithrix (sic!) (Common Marmoset,
Callithrix (Callithrix) jacchus; Black-tufted Marmoset, Callithrix
(Callithrix) penicillata; Wied's Marmoset, Callithrix (Callithrix)
kuhlii; White-headed Marmoset, Callithrix (Callithrix) geoffroyi;
Buffy-headed Marmoset, Callithrix (Callithrix) flaviceps;
Buffy-tufted Marmoset, Callithrix (Callithrix) aurita); belonging
to the Amazonian marmosets of subgenus Mico (Rio Acari Marmoset,
Callithrix (Mico) acariensis; Manicore Marmoset, Callithrix (Mico)
manicorensis; Silvery Marmoset, Callithrix (Mico) argentata; White
Marmoset, Callithrix (Mico) leucippe; Emilia's Marmoset, Callithrix
(Mico) emiliae; Black-headed Marmoset, Callithrix (Mico) nigriceps;
Marca's Marmoset, Callithrix (Mico)marcai; Black-tailed Marmoset,
Callithrix (Mico) melanura; Santarem Marmoset, Callithrix (Mico)
humeralifera; Maues Marmoset, Callithrix (Mico) mauesi;
Gold-and-white Marmoset, Callithrix (Mico) chrysoleuca;
Hershkovitz's Marmoset, Callithrix (Mico) intermedia; l Satere
Marmoset, Callithrix (Mico) saterei); Roosmalens' Dwarf Marmoset
belonging to the subgenus Callibella (Callithrix (Callibella)
humilis); or the Pygmy Marmoset belonging to the subgenus Cebuella
(Callithrix (Cebuella) pygmaea).
[0071] Other genera of the New-World Monkeys comprise tamarins of
the genus Saguinus (comprising the S. oedipus-group, the S. midas
group, the S. nigricollis group, the S. mystax group, the S.
bicolor group and the S. inustus group) and squirrel monkeys of the
genus Samiri (e.g. Saimiri sciureus, Saimiri oerstedii, Saimiri
ustus, Saimiri boliviensis, Saimiri vanzolini)
[0072] In a preferred embodiment of the bispecific single chain
antibody molecule of the invention, the non-chimpanzee primate is
an old world monkey. In a more preferred embodiment of the
polypeptide, the old world monkey is a monkey of the Papio genus
Macaque genus. Most preferably, the monkey of the Macaque genus is
Assamese macaque (Macaca assamensis), Barbary macaque (Macaca
sylvanus), Bonnet macaque (Macaca radiata), Booted or
Sulawesi-Booted macaque (Macaca ochreata), Sulawesi-crested macaque
(Macaca nigra), Formosan rock macaque (Macaca cyclopsis), Japanese
snow macaque or Japanese macaque (Macaca fuscata), Cynomologus
monkey or crab-eating macaque or long-tailed macaque or Java
macaque (Macaca fascicularis), Lion-tailed macaque (Macaca
silenus), Pigtailed macaque (Macaca nemestrina), Rhesus macaque
(Macaca mulatta), Tibetan macaque (Macaca thibetana), Tonkean
macaque (Macaca tonkeana), Toque macaque (Macaca sinica),
Stump-tailed macaque or Red-faced macaque or Bear monkey (Macaca
arctoides), or Moor macaque (Macaca maurus). Most preferably, the
monkey of the Papio genus is Hamadryas Baboon, Papio hamadryas;
Guinea Baboon, Papio papio; Olive Baboon, Papio anubis; Yellow
Baboon, Papio cynocephalus; Chacma Baboon, Papio ursinus.
[0073] In an alternatively preferred embodiment of the bispecific
single chain antibody molecule of the invention, the non-chimpanzee
primate is a new world monkey. In a more preferred embodiment of
the polypeptide, the new world monkey is a monkey of the Callithrix
genus (marmoset), the Saguinus genus or the Samiri genus. Most
preferably, the monkey of the Callithrix genus is Callithrix
jacchus, the monkey of the Saguinus genus is Saguinus oedipus and
the monkey of the Samiri genus is Saimiri sciureus.
[0074] The term "cell surface antigen" as used herein denotes a
molecule, which is displayed on the surface of a cell. In most
cases, this molecule will be located in or on the plasma membrane
of the cell such that at least part of this molecule remains
accessible from outside the cell in tertiary form. A non-limiting
example of a cell surface molecule, which is located in the plasma
membrane is a transmembrane protein comprising, in its tertiary
conformation, regions of hydrophilicity and hydrophobicity. Here,
at least one hydrophobic region allows the cell surface molecule to
be embedded, or inserted in the hydrophobic plasma membrane of the
cell while the hydrophilic regions extend on either side of the
plasma membrane into the cytoplasm and extracellular space,
respectively. Non-limiting examples of cell surface molecules which
are located on the plasma membrane are proteins which have been
modified at a cysteine residue to bear a palmitoyl group, proteins
modified at a C-terminal cysteine residue to bear a farnesyl group
or proteins which have been modified at the C-terminus to bear a
glycosyl phosphatidyl inositol ("GPI") anchor. These groups allow
covalent attachment of proteins to the outer surface of the plasma
membrane, where they remain accessible for recognition by
extracellular molecules such as antibodies. Examples of cell
surface antigens are CD3 epsilon and PSMA. As described herein
above, PSMA is a cell surface antigen which is a target for therapy
of cancer, including, but not limited to solid tumors, preferably
carcinomas and prostate cancer.
[0075] In light of this, PSMA can also be characterized as a tumor
antigen. The term "tumor antigen" as used herein may be understood
as those antigens that are presented on tumor cells. These antigens
can be presented on the cell surface with an extracellular part,
which is often combined with a transmembrane and cytoplasmic part
of the molecule. These antigens can sometimes be presented only by
tumor cells and never by the normal ones. Tumor antigens can be
exclusively expressed on tumor cells or might represent a tumor
specific mutation compared to normal cells. In this case, they are
called tumor-specific antigens. More common are antigens that are
presented by tumor cells and normal cells, and they are called
tumor-associated antigens. These tumor-associated antigens can be
overexpressed compared to normal cells or are accessible for
antibody binding in tumor cells due to the less compact structure
of the tumor tissue compared to normal tissue. One example for a
tumor antigen in line with the present invention is PSMA.
[0076] As described herein above the bispecific single chain
antibody molecule of the invention binds with the first binding
domain to an epitope of human and non-chimpanzee primate
CD3.epsilon. (epsilon) chain, wherein the epitope is part of an
amino acid sequence comprised in the group consisting of 27 amino
acid residues as depicted in SEQ ID NOs. 2, 4, 6, or 8 or a
functional fragment thereof.
[0077] In line with the present invention it is preferred for the
bispecific single chain antibody molecule of the invention that
said epitope is part of an amino acid sequence comprising 26, 25,
24, 23, 22, 21, 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8,
7, 6 or 5 amino acids.
[0078] More preferably, wherein said epitope comprises at least the
amino acid sequence Gln-Asp-Gly-Asn-Glu (Q-D-G-N-E).
[0079] Within the present invention, a functional fragment of the
N-terminal 1-27 amino acid residues means that said functional
fragment is still a context-independent epitope maintaining its
three-dimensional structural integrity when taken out of its native
environment in the CD3 complex (and fused to a heterologous amino
acid sequence such as EpCAM or an immunoglobulin Fc part, e.g. as
shown in Example 3.1). The maintenance of the three-dimensional
structure within the 27 amino acid N-terminal polypeptide or
functional fragment thereof of CD3 epsilon can be used for the
generation of binding domains which bind to the N-terminal CD3
epsilon polypeptide fragment in vitro and to the native (CD3
epsilon subunit of the) CD3 complex on T cells in vivo with the
same binding affinity. Within the present invention, a functional
fragment of the N-terminal 1-27 amino acid residues means that CD3
binding domains provided herein can still bind to such functional
fragments in a context-independent manner. The person skilled in
the art is aware of methods for epitope mapping to determine which
amino acid residues of an epitope are recognized by such anti-CD3
binding domains (e.g. alanine scanning; see appended examples).
[0080] In one embodiment of the invention, the bispecific single
chain antibody molecule of the invention comprises a (first)
binding domain capable of binding to an epitope of human and
non-chimpanzee primate CD3.epsilon. chain and a second binding
domain capable of binding to the cell surface antigen PSMA.
[0081] Within the present invention it is further preferred that
the second binding domain binds to the human cell surface antigen
PSMA and/or a non-chimpanzee primate PSMA. Particularly preferred,
the second binding domain binds to the human PSMA and a
non-chimpanzee primate PSMA, preferably a macaque PSMA. It is to be
understood, that the second binding domain binds to at least one
non-chimpanzee primate PSMA, however, it may also bind to two,
three or more, non-chimpanzee primate PSMA homologs. For example,
the second binding domain may bind to the Cynomolgus monkey PSMA
and to the Rhesus monkey PSMA.
[0082] The present invention including all methods, uses, kits etc.
described herein, also relates to the seconed binding domains as
such (i.e. not in the context of a bispecific single chain
antibody). "As such" further includes antibody formats other than
the bispecific single chain antibodies as described herein, for
example antibody fragments (comprising the second domain),
humanized antibodies, fusion proteins comprising the second domain
etc. Antibody formats other than the bispecific single chain
antibodies of the present invention are also described herein
above.
[0083] For the generation of the second binding domain of the
bispecific single chain antibody molecule of the invention, e.g.
bispecific single chain antibodies as defined herein, monoclonal
antibodies binding to both of the respective human and/or
non-chimpanzee primate cell surface antigen such as PSMA can be
utilized. Appropriate binding domains for the bispecific
polypeptide as defined herein e.g. can be derived from
cross-species specific monoclonal antibodies by recombinant methods
described in the art. A monoclonal antibody binding to a human cell
surface antigen and to the homolog of said cell surface antigen in
a non-chimpanzee primate can be tested by FACS assays as set forth
above. It is evident to those skilled in the art that cross-species
specific antibodies can also be generated by hybridoma techniques
described in the literature (Milstein and Kohler, Nature 256
(1975), 495-7). For example, mice may be alternately immunized with
human and non-chimpanzee primate cell surface antigen, such as
PSMA. From these mice, cross-species specific antibody-producing
hybridoma cells are isolated via hybridoma technology and analysed
by FACS as set forth above. The generation and analysis of
bispecific polypeptides such as bispecific single chain antibodies
exhibiting cross-species specificity as described herein is shown
in the following examples. The advantages of the bispecific single
chain antibodies exhibiting cross-species specificity include the
points enumerated herein.
[0084] It is particularly preferred for the bispecific single chain
antibody molecule of the invention that the first binding domain
capable of binding to an epitope of human and non-chimpanzee
primate CD3.epsilon. chain comprises a VL region comprising CDR-L1,
CDR-L2 and CDR-L3 selected from: [0085] (a) CDR-L1 as depicted in
SEQ ID NO. 27, CDR-L2 as depicted in SEQ ID NO. 28 and CDR-L3 as
depicted in SEQ ID NO. 29; [0086] (b) CDR-L1 as depicted in SEQ ID
NO. 117, CDR-L2 as depicted in SEQ ID NO. 118 and CDR-L3 as
depicted in SEQ ID NO. 119; and [0087] (c) CDR-L1 as depicted in
SEQ ID NO. 153, CDR-L2 as depicted in SEQ ID NO. 154 and CDR-L3 as
depicted in SEQ ID NO. 155.
[0088] The variable regions, i.e. the variable light chain ("L" or
"VL") and the variable heavy chain ("H" or "VH") are understood in
the art to provide the binding domain of an antibody. This variable
regions harbor the complementary determining regions.
[0089] The term "complementary determining region" (CDR) is well
known in the art to dictate the antigen specificity of an antibody.
The term "CDR-L" or "L CDR" or "LCDR" refers to CDRs in the VL,
whereas the term "CDR-H" or "H CDR" or HCDR'' refers to the CDRs in
the VH.
[0090] In an alternatively preferred embodiment of the bispecific
single chain antibody molecule of the invention the first binding
domain capable of binding to an epitope of human and non-chimpanzee
primate CD3.epsilon. chain comprises a VH region comprising CDR-H
1, CDR-H2 and CDR-H3 selected from: [0091] (a) CDR-H1 as depicted
in SEQ ID NO. 12, CDR-H2 as depicted in SEQ ID NO. 13 and CDR-H3 as
depicted in SEQ ID NO. 14; [0092] (b) CDR-H1 as depicted in SEQ ID
NO. 30, CDR-H2 as depicted in SEQ ID NO. 31 and CDR-H3 as depicted
in SEQ ID NO. 32; [0093] (c) CDR-H1 as depicted in SEQ ID NO. 48,
CDR-H2 as depicted in SEQ ID NO. 49 and CDR-H3 as depicted in SEQ
ID NO. 50; [0094] (d) CDR-H1 as depicted in SEQ ID NO. 66, CDR-H2
as depicted in SEQ ID NO. 67 and CDR-H3 as depicted in SEQ ID NO.
68; [0095] (e) CDR-H1 as depicted in SEQ ID NO. 84, CDR-H2 as
depicted in SEQ ID NO. 85 and CDR-H3 as depicted in SEQ ID NO. 86;
[0096] (f) CDR-H1 as depicted in SEQ ID NO. 102, CDR-H2 as depicted
in SEQ ID NO. 103 and CDR-H3 as depicted in SEQ ID NO. 104; [0097]
(g) CDR-H1 as depicted in SEQ ID NO. 120, CDR-H2 as depicted in SEQ
ID NO. 121 and CDR-H3 as depicted in SEQ ID NO. 122; [0098] (h)
CDR-H1 as depicted in SEQ ID NO. 138, CDR-H2 as depicted in SEQ ID
NO. 139 and CDR-H3 as depicted in SEQ ID NO. 140; [0099] (i) CDR-H1
as depicted in SEQ ID NO. 156, CDR-H2 as depicted in SEQ ID NO. 157
and CDR-H3 as depicted in SEQ ID NO. 158; and [0100] (j) CDR-H1 as
depicted in SEQ ID NO. 174, CDR-H2 as depicted in SEQ ID NO. 175
and CDR-H3 as depicted in SEQ ID NO. 176.
[0101] It is further preferred that the binding domain capable of
binding to an epitope of human and non-chimpanzee primate
CD3.epsilon. chain comprises a VL region selected from the group
consisting of a VL region as depicted in SEQ ID NO. 35, 39, 125,
129, 161 or 165.
[0102] It is alternatively preferred that the first binding domain
capable of binding to an epitope of human and non-chimpanzee
primate CD3.epsilon. chain comprises a VH region selected from the
group consisting of a VH region as depicted in SEQ ID NO. 15, 19,
33, 37, 51, 55, 69, 73, 87, 91, 105, 109, 123, 127, 141, 145, 159,
163, 177 or 181.
[0103] More preferably, the bispecific single chain antibody
molecule of the invention is characterized by the first binding
domain capable of binding to an epitope of human and non-chimpanzee
primate CD3.epsilon. chain, which comprises a VL region and a VH
region selected from the group consisting of: [0104] (a) a VL
region as depicted in SEQ ID NO. 17 or 21 and a VH region as
depicted in SEQ ID NO. 15 or 19; [0105] (b) a VL region as depicted
in SEQ ID NO. 35 or 39 and a VH region as depicted in SEQ ID NO. 33
or 37; [0106] (c) a VL region as depicted in SEQ ID NO. 53 or 57
and a VH region as depicted in SEQ ID NO. 51 or 55; [0107] (d) a VL
region as depicted in SEQ ID NO. 71 or 75 and a VH region as
depicted in SEQ ID NO. 69 or 73; [0108] (e) a VL region as depicted
in SEQ ID NO. 89 or 93 and a VH region as depicted in SEQ ID NO. 87
or 91; [0109] (f) a VL region as depicted in SEQ ID NO. 107 or 111
and a VH region as depicted in SEQ ID NO. 105 or 109; [0110] (g) a
VL region as depicted in SEQ ID NO. 125 or 129 and a VH region as
depicted in SEQ ID NO. 123 or 127; [0111] (h) a VL region as
depicted in SEQ ID NO. 143 or 147 and a VH region as depicted in
SEQ ID NO. 141 or 145; [0112] (i) a VL region as depicted in SEQ ID
NO. 161 or 165 and a VH region as depicted in SEQ ID NO. 159 or
163; and [0113] (j) a VL region as depicted in SEQ ID NO. 179 or
183 and a VH region as depicted in SEQ ID NO. 177 or 181.
[0114] According to a preferred embodiment of the bispecific single
chain antibody molecule of the invention the pairs of VH-regions
and VL-regions in the first binding domain binding to CD3 epsilon
are in the format of a single chain antibody (scFv). The VH and VL
regions are arranged in the order VH-VL or VL-VH. It is preferred
that the VH-region is positioned N-terminally to a linker sequence.
The VL-region is positioned C-terminally of the linker sequence.
Put in other words, the domain arrangement in the CD3 binding
domain of the bispecific single chain antibody molecule of the
invention is preferably VH-VL, with said CD3 binding domain located
C-terminally to the second (cell surface antigen, such as PSMA)
binding domain. Preferably the VH-VL comprises or is SEQ ID NO.
185.
[0115] A preferred embodiment of the above described bispecific
single chain antibody molecule of the invention is characterized by
the first binding domain capable of binding to an epitope of human
and non-chimpanzee primate CD3.epsilon. chain comprising an amino
acid sequence selected from the group consisting of SEQ ID NOs: 23,
25, 41, 43, 59, 61, 77, 79, 95, 97, 113, 115, 131, 133, 149, 151,
167, 169, 185 or 187.
[0116] The invention further relates to an above described
bispecific single chain antibody, wherein the second binding domain
binds to the cell surface antigen PSMA.
[0117] According to a preferred embodiment of the invention an
above characterized bispecific single chain antibody molecule
comprises a group of the following sequences as CDR H1, CDR H2, CDR
H3, CDR L1, CDR L2 and CDR L3 in the second binding domain selected
from the group consisting of: [0118] a) CDR H1-3 of SEQ ID NO:
394-396 and CDR L1-3 of SEQ ID NO: 389-391; [0119] b) CDR H1-3 of
SEQ ID NO: 408-410 and CDR L1-3 of SEQ ID NO: 403-405; [0120] c)
CDR H1-3 of SEQ ID NO: 422-424 and CDR L1-3 of SEQ ID NO: 417-419;
[0121] d) CDR H1-3 of SEQ ID NO: 436-438 and CDR L1-3 of SEQ ID NO:
431-433; [0122] e) CDR H1-3 of SEQ ID NO: 445-447 and CDR L1-3 of
SEQ ID NO: 450-452; [0123] f) CDR H1-3 of SEQ ID NO: 464-466 and
CDR L1-3 of SEQ ID NO: 459-461; [0124] g) CDR H1-3 of SEQ ID NO:
478-480 and CDR L1-3 of SEQ ID NO: 473-475; [0125] h) CDR H1-3 of
SEQ ID NO: 492-494 and CDR L1-3 of SEQ ID NO: 487-489; [0126] i)
CDR H1-3 of SEQ ID NO: 506-508 and CDR L1-3 of SEQ ID NO: 501-503;
[0127] j) CDR H1-3 of SEQ ID NO: 520-522 and CDR L1-3 of SEQ ID NO:
515-517; [0128] k) CDR H1-3 of SEQ ID NO: 534-536 and CDR L1-3 of
SEQ ID NO: 529-531; [0129] l) CDR H1-3 of SEQ ID NO: 548-550 and
CDR L1-3 of SEQ ID NO: 543-545; [0130] m) CDR H1-3 of SEQ ID NO:
562-564 and CDR L1-3 of SEQ ID NO: 557-559; [0131] n) CDR H1-3 of
SEQ ID NO: 576-578 and CDR L1-3 of SEQ ID NO: 571-573; [0132] o)
CDR H1-3 of SEQ ID NO: 590-592 and CDR L1-3 of SEQ ID NO: 585-587;
[0133] p) CDR H1-3 of SEQ ID NO: 604-606 and CDR L1-3 of SEQ ID NO:
599-601; [0134] q) CDR H1-3 of SEQ ID NO: 618-620 and CDR L1-3 of
SEQ ID NO: 613-615; [0135] r) CDR H1-3 of SEQ ID NO: 632-634 and
CDR L1-3 of SEQ ID NO: 627-629; [0136] s) CDR H1-3 of SEQ ID NO:
646-648 and CDR L1-3 of SEQ ID NO: 641-643; [0137] t) CDR H1-3 of
SEQ ID NO: 660-662 and CDR L1-3 of SEQ ID NO: 655-657; [0138] u)
CDR H1-3 of SEQ ID NO: 674-676 and CDR L1-3 of SEQ ID NO: 669-671;
[0139] v) CDR H1-3 of SEQ ID NO: 688-690 and CDR L1-3 of SEQ ID NO:
683-685; [0140] w) CDR H1-3 of SEQ ID NO: 702-704 and CDR L1-3 of
SEQ ID NO: 697-699; [0141] x) CDR H1-3 of SEQ ID NO: 716-718 and
CDR L1-3 of SEQ ID NO: 711-713; and [0142] y) CDR H1-3 of SEQ ID
NO: 729-731 and CDR L1-3 of SEQ ID NO: 724-726.
[0143] The sequences of the corresponding VL- and VH-regions of the
second binding domain of the bispecific single chain antibody
molecule of the invention as well as of the respective scFvs are
shown in the sequence listing.
[0144] In the bispecific single chain antibody molecule of the
invention the binding domains are arranged in the order
VL-VH-VH-VL, VL-VH-VL-VH, VH-VL-VH-VL or VH-VL-VL-VH, as
exemplified in the appended examples. Preferably, the binding
domains are arranged in the order VH PSMA-VL PSMA-VH CD3-VL CD3 or
VL PSMA-VH PSMA-VH CD3-VL CD3.
[0145] A particularly preferred embodiment of the invention
concerns an above characterized polypeptide, wherein the bispecific
single chain antibody molecule comprises a sequence selected from:
[0146] (a) an amino acid sequence as depicted in any of SEQ ID NOs:
399, 413, 427, 441, 455, 469, 483, 497, 511, 525, 539, 553, 567,
581, 595, 609, 623, 637, 651, 665, 679, 693, 707, 721, 734, 799,
817, 863, 849, 835, 785, 899, 935, 1017, 1031, 917, 1003, 953, 971
or 989; [0147] (b) an amino acid sequence encoded by a nucleic acid
sequence as depicted in any of SEQ ID NOs: 400, 414, 428, 442, 456,
470, 484, 498, 512, 526, 540, 554, 568, 582, 596, 610, 624, 638,
652, 666, 680, 694, 708, 736 735, 800, 818, 864, 850, 836, 786,
882, 900, 936, 1018, 1032, 918, 1004, 954, 972, 990, 804, 822, 868,
886, 904, 940, 922, 958 or 976; [0148] (c) an amino acid sequence
at least 90% identical, more preferred at least 95% identical, most
preferred at least 96% identical to the amino acid sequence of (a)
or (b).
[0149] The invention relates to a bispecific single chain antibody
molecule comprising an amino acid sequence as depicted in any of
SEQ ID NOs: 399, 413, 427, 441, 455, 469, 483, 497, 511, 525, 539,
553, 567, 581, 595, 609, 623, 637, 651, 665, 679, 693, 707, 721,
734, 799, 817, 863, 849, 835, 785, 899, 935, 1017, 1031, 917, 1003,
953, 971 or 989, as well as to amino acid sequences at least 85%
identical, preferably 90%, more preferred at least 95% identical,
most preferred at least 96, 97, 98, or 99% identical to the amino
acid sequence of SEQ ID NOs: 399, 413, 427, 441, 455, 469, 483,
497, 511, 525, 539, 553, 567, 581, 595, 609, 623, 637, 651, 665,
679, 693, 707, 721, 734, 799, 817, 863, 849, 835, 785, 899, 935,
1017, 1031, 917, 1003, 953, 971 or 989. The invention relates also
to the corresponding nucleic acid sequences as depicted in any of
SEQ ID NOs: 400, 414, 428, 442, 456, 470, 484, 498, 512, 526, 540,
554, 568, 582, 596, 610, 624, 638, 652, 666, 680, 694, 708, 736
735, 800, 818, 864, 850, 836, 786, 882, 900, 936, 1018, 1032, 918,
1004, 954, 972, 990, 804, 822, 868, 886, 904, 940, 922, 958 or 976
as well as to nucleic acid sequences at least 85% identical,
preferably 90%, more preferred at least 95% identical, most
preferred at least 96, 97, 98, or 99% identical to the nucleic acid
sequences shown in SEQ ID NOs: 400, 414, 428, 442, 456, 470, 484,
498, 512, 526, 540, 554, 568, 582, 596, 610, 624, 638, 652, 666,
680, 694, 708, 736, 735, 800, 818, 864, 850, 836, 786, 882, 900,
936, 1018, 1032, 918, 1004, 954, 972, 990, 804, 822, 868, 886, 904,
940, 922, 958 or 976. It is to be understood that the sequence
identity is determined over the entire nucleotide or amino acid
sequence. For sequence alignments, for example, the programs Gap or
BestFit can be used (Needleman and Wunsch J. Mol. Biol. 48 (1970),
443-453; Smith and Waterman, Adv. Appl. Math 2 (1981), 482-489),
which is contained in the GCG software package (Genetics Computer
Group, 575 Science Drive, Madison, Wis., USA 53711 (1991). It is a
routine method for those skilled in the art to determine and
identify a nucleotide or amino acid sequence having e.g. 85% (90%,
95%, 96%, 97%, 98% or 99%) sequence identity to the nucleotide or
amino acid sequences of the bispecific single chain antibody of the
invention by using e.g. one of the above mentioned programs. For
example, according to Crick's Wobble hypothesis, the 5' base on the
anti-codon is not as spatially confined as the other two bases, and
could thus have non-standard base pairing. Put in other words: the
third position in a codon triplet may vary so that two triplets
which differ in this third position may encode the same amino acid
residue. Said hypothesis is well known to the person skilled in the
art (see e.g.--http_colon-slash-slash
en.wikipedia.org/wiki/Wobble_Hypothesis; Crick, J Mol Biol 19
(1966): 548-55).
[0150] Preferred domain arrangements in the PSMA.times.CD3
bispecific single chain antibody constructs of the invention are
shown in the following examples.
[0151] In a preferred embodiment of the invention, the bispecific
single chain antibodies are cross-species specific for CD3 epsilon
and for the human and non-chimpanzee primate cell surface antigen
PSMA, recognized by their second binding domain.
[0152] In an alternative embodiment the present invention provides
a nucleic acid sequence encoding an above described bispecific
single chain antibody molecule of the invention.
[0153] The present invention also relates to a vector comprising
the nucleic acid molecule of the present invention.
[0154] Many suitable vectors are known to those skilled in
molecular biology, the choice of which would depend on the function
desired and include plasmids, cosmids, viruses, bacteriophages and
other vectors used conventionally in genetic engineering. Methods
which are well known to those skilled in the art can be used to
construct various plasmids and vectors; see, for example, the
techniques described in Sambrook et al. (loc cit.) and Ausubel,
Current Protocols in Molecular Biology, Green Publishing Associates
and Wiley Interscience, N.Y. (1989), (1994). Alternatively, the
polynucleotides and vectors of the invention can be reconstituted
into liposomes for delivery to target cells. As discussed in
further details below, a cloning vector was used to isolate
individual sequences of DNA. Relevant sequences can be transferred
into expression vectors where expression of a particular
polypeptide is required. Typical cloning vectors include
pBluescript SK, pGEM, pUC9, pBR322 and pGBT9. Typical expression
vectors include pTRE, pCAL-n-EK, pESP-1, pPOP13CAT.
[0155] Preferably said vector comprises a nucleic acid sequence
which is a regulatory sequence operably linked to said nucleic acid
sequence defined herein.
[0156] The term "regulatory sequence" refers to DNA sequences,
which are necessary to effect the expression of coding sequences to
which they are ligated. The nature of such control sequences
differs depending upon the host organism. In prokaryotes, control
sequences generally include promoter, ribosomal binding site, and
terminators. In eukaryotes generally control sequences include
promoters, terminators and, in some instances, enhancers,
transactivators or transcription factors. The term "control
sequence" is intended to include, at a minimum, all components the
presence of which are necessary for expression, and may also
include additional advantageous components.
[0157] The term "operably linked" refers to a juxtaposition wherein
the components so described are in a relationship permitting them
to function in their intended manner. A control sequence "operably
linked" to a coding sequence is ligated in such a way that
expression of the coding sequence is achieved under conditions
compatible with the control sequences. In case the control sequence
is a promoter, it is obvious for a skilled person that
double-stranded nucleic acid is preferably used.
[0158] Thus, the recited vector is preferably an expression vector.
An "expression vector" is a construct that can be used to transform
a selected host and provides for expression of a coding sequence in
the selected host. Expression vectors can for instance be cloning
vectors, binary vectors or integrating vectors. Expression
comprises transcription of the nucleic acid molecule preferably
into a translatable mRNA. Regulatory elements ensuring expression
in prokaryotes and/or eukaryotic cells are well known to those
skilled in the art. In the case of eukaryotic cells they comprise
normally promoters ensuring initiation of transcription and
optionally poly-A signals ensuring termination of transcription and
stabilization of the transcript. Possible regulatory elements
permitting expression in prokaryotic host cells comprise, e.g., the
P.sub.L, lac, trp or tac promoter in E. coli, and examples of
regulatory elements permitting expression in eukaryotic host cells
are the AOX1 or GAL1 promoter in yeast or the CMV-, SV40-,
RSV-promoter (Rous sarcoma virus), CMV-enhancer, SV40-enhancer or a
globin intron in mammalian and other animal cells.
[0159] Beside elements, which are responsible for the initiation of
transcription such regulatory elements may also comprise
transcription termination signals, such as the SV40-poly-A site or
the tk-poly-A site, downstream of the polynucleotide. Furthermore,
depending on the expression system used leader sequences capable of
directing the polypeptide to a cellular compartment or secreting it
into the medium may be added to the coding sequence of the recited
nucleic acid sequence and are well known in the art; see also the
appended Examples. The leader sequence(s) is (are) assembled in
appropriate phase with translation, initiation and termination
sequences, and preferably, a leader sequence capable of directing
secretion of translated protein, or a portion thereof, into the
periplasmic space or extracellular medium. Optionally, the
heterologous sequence can encode a fusion protein including an
N-terminal identification peptide imparting desired
characteristics, e.g., stabilization or simplified purification of
expressed recombinant product; see supra. In this context, suitable
expression vectors are known in the art such as Okayama-Berg cDNA
expression vector pcDV1 (Pharmacia), pCDM8, pRc/CMV, pcDNA1, pcDNA3
(In-vitrogene), pEF-DHFR, pEF-ADA or pEF-neo (Mack et al. PNAS
(1995) 92, 7021-7025 and Raum et al. Cancer Immunol Immunother
(2001) 50(3), 141-150) or pSPORT1 (GIBCO BRL).
[0160] Preferably, the expression control sequences will be
eukaryotic promoter systems in vectors capable of transforming of
transfecting eukaryotic host cells, but control sequences for
prokaryotic hosts may also be used. Once the vector has been
incorporated into the appropriate host, the host is maintained
under conditions suitable for high level expression of the
nucleotide sequences, and as desired, the collection and
purification of the bispecific single chain antibody molecule of
the invention may follow; see, e.g., the appended examples.
[0161] An alternative expression system, which can be used to
express a cell cycle interacting protein is an insect system. In
one such system, Autographa californica nuclear polyhedrosis virus
(AcNPV) is used as a vector to express foreign genes in Spodoptera
frugiperda cells or in Trichoplusia larvae. The coding sequence of
a recited nucleic acid molecule may be cloned into a nonessential
region of the virus, such as the polyhedrin gene, and placed under
control of the polyhedrin promoter. Successful insertion of said
coding sequence will render the polyhedrin gene inactive and
produce recombinant virus lacking coat protein coat. The
recombinant viruses are then used to infect S. frugiperda cells or
Trichoplusia larvae in which the protein of the invention is
expressed (Smith, J. Virol. 46 (1983), 584; Engelhard, Proc. Nat.
Acad. Sci. USA 91 (1994), 3224-3227).
[0162] Additional regulatory elements may include transcriptional
as well as translational enhancers. Advantageously, the
above-described vectors of the invention comprise a selectable
and/or scorable marker.
[0163] Selectable marker genes useful for the selection of
transformed cells and, e.g., plant tissue and plants are well known
to those skilled in the art and comprise, for example,
antimetabolite resistance as the basis of selection for dhfr, which
confers resistance to methotrexate (Reiss, Plant Physiol. (Life
Sci. Adv.) 13 (1994), 143-149); npt, which confers resistance to
the aminoglycosides neomycin, kanamycin and paromycin
(Herrera-Estrella, EMBO J. 2 (1983), 987-995) and hygro, which
confers resistance to hygromycin (Marsh, Gene 32 (1984), 481-485).
Additional selectable genes have been described, namely trpB, which
allows cells to utilize indole in place of tryptophan; hisD, which
allows cells to utilize histinol in place of dine (Hartman, Proc.
Natl. Acad. Sci. USA 85 (1988), 8047); mannose-6-phosphate
isomerase which allows cells to utilize mannose (WO 94/20627) and
ODC (ornithine decarboxylase) which confers resistance to the
ornithine decarboxylase inhibitor, 2-(difluoromethyl)-DL-ornithine,
DFMO (McConlogue, 1987, In: Current Communications in Molecular
Biology, Cold Spring Harbor Laboratory ed.) or deaminase from
Aspergillus terreus which confers resistance to Blasticidin S
(Tamura, Biosci. Biotechnol. Biochem. 59 (1995), 2336-2338).
[0164] Useful scorable markers are also known to those skilled in
the art and are commercially available. Advantageously, said marker
is a gene encoding luciferase (Giacomin, Pl. Sci. 116 (1996),
59-72; Scikantha, J. Bact. 178 (1996), 121), green fluorescent
protein (Gerdes, FEBS Lett. 389 (1996), 44-47) or
.beta.-glucuronidase (Jefferson, EMBO J. 6 (1987), 3901-3907). This
embodiment is particularly useful for simple and rapid screening of
cells, tissues and organisms containing a recited vector.
[0165] As described above, the recited nucleic acid molecule can be
used alone or as part of a vector to express the bispecific single
chain antibody molecule of the invention in cells, for, e.g.,
purification but also for gene therapy purposes. The nucleic acid
molecules or vectors containing the DNA sequence(s) encoding any
one of the above described bispecific single chain antibody
molecule of the invention is introduced into the cells which in
turn produce the polypeptide of interest. Gene therapy, which is
based on introducing therapeutic genes into cells by ex-vivo or
in-vivo techniques is one of the most important applications of
gene transfer. Suitable vectors, methods or gene-delivery systems
for in-vitro or in-vivo gene therapy are described in the
literature and are known to the person skilled in the art; see,
e.g., Giordano, Nature Medicine 2 (1996), 534-539; Schaper, Circ.
Res. 79 (1996), 911-919; Anderson, Science 256 (1992), 808-813;
Verma, Nature 389 (1994), 239; Isner, Lancet 348 (1996), 370-374;
Muhlhauser, Circ. Res. 77 (1995), 1077-1086; Onodera, Blood 91
(1998), 30-36; Verma, Gene Ther. 5 (1998), 692-699; Nabel, Ann.
N.Y. Acad. Sci. 811 (1997), 289-292; Verzeletti, Hum. Gene Ther. 9
(1998), 2243-51; Wang, Nature Medicine 2 (1996), 714-716; WO
94/29469; WO 97/00957, U.S. Pat. Nos. 5,580,859; 5,589,466; or
Schaper, Current Opinion in Biotechnology 7 (1996), 635-640; dos
Santos Coura and Nardi Virol J. (2007), 4:99. The recited nucleic
acid molecules and vectors may be designed for direct introduction
or for introduction via liposomes, or viral vectors (e.g.,
adenoviral, retroviral) into the cell. Preferably, said cell is a
germ line cell, embryonic cell, or egg cell or derived there from,
most preferably said cell is a stem cell. An example for an
embryonic stem cell can be, inter alia, a stem cell as described in
Nagy, Proc. Natl. Acad. Sci. USA 90 (1993), 8424-8428.
[0166] The invention also provides for a host transformed or
transfected with a vector of the invention. Said host may be
produced by introducing the above described vector of the invention
or the above described nucleic acid molecule of the invention into
the host. The presence of at least one vector or at least one
nucleic acid molecule in the host may mediate the expression of a
gene encoding the above described single chain antibody
constructs.
[0167] The described nucleic acid molecule or vector of the
invention, which is introduced in the host may either integrate
into the genome of the host or it may be maintained
extrachromosomally.
[0168] The host can be any prokaryote or eukaryotic cell.
[0169] The term "prokaryote" is meant to include all bacteria,
which can be transformed or transfected with DNA or RNA molecules
for the expression of a protein of the invention. Prokaryotic hosts
may include gram negative as well as gram positive bacteria such
as, for example, E. coli, S. typhimurium, Serratia marcescens and
Bacillus subtilis. The term "eukaryotic" is meant to include yeast,
higher plant, insect and preferably mammalian cells. Depending upon
the host employed in a recombinant production procedure, the
protein encoded by the polynucleotide of the present invention may
be glycosylated or may be non-glycosylated. Especially preferred is
the use of a plasmid or a virus containing the coding sequence of
the bispecific single chain antibody molecule of the invention and
genetically fused thereto an N-terminal FLAG-tag and/or C-terminal
His-tag. Preferably, the length of said FLAG-tag is about 4 to 8
amino acids, most preferably 8 amino acids. An above described
polynucleotide can be used to transform or transfect the host using
any of the techniques commonly known to those of ordinary skill in
the art. Furthermore, methods for preparing fused, operably linked
genes and expressing them in, e.g., mammalian cells and bacteria
are well-known in the art (Sambrook, loc cit.). Preferably, said
the host is a bacterium or an insect, fungal, plant or animal cell.
It is particularly envisaged that the recited host may be a
mammalian cell. Particularly preferred host cells comprise CHO
cells, COS cells, myeloma cell lines like SP2/0 or NS/0. As
illustrated in the appended examples, particularly preferred are
CHO-cells as hosts.
[0170] More preferably said host cell is a human cell or human cell
line, e.g. per.c6 (Kroos, Biotechnol. Prog., 2003, 19:163-168).
[0171] In a further embodiment, the present invention thus relates
to a process for the production of a bispecific single chain
antibody molecule of the invention, said process comprising
culturing a host of the invention under conditions allowing the
expression of the bispecific single chain antibody molecule of the
invention and recovering the produced polypeptide from the
culture.
[0172] The transformed hosts can be grown in fermentors and
cultured according to techniques known in the art to achieve
optimal cell growth. The bispecific single chain antibody molecule
of the invention can then be isolated from the growth medium,
cellular lysates, or cellular membrane fractions. The isolation and
purification of the, e.g., microbially expressed bispecific single
chain antibody molecules may be by any conventional means such as,
for example, preparative chromatographic separations and
immunological separations such as those involving the use of
monoclonal or polyclonal antibodies directed, e.g., against a tag
of the bispecific single chain antibody molecule of the invention
or as described in the appended examples.
[0173] The conditions for the culturing of a host, which allow the
expression are known in the art to depend on the host system and
the expression system/vector used in such process. The parameters
to be modified in order to achieve conditions allowing the
expression of a recombinant polypeptide are known in the art. Thus,
suitable conditions can be determined by the person skilled in the
art in the absence of further inventive input.
[0174] Once expressed, the bispecific single chain antibody
molecule of the invention can be purified according to standard
procedures of the art, including ammonium sulfate precipitation,
affinity columns, column chromatography, gel electrophoresis and
the like; see, Scopes, "Protein Purification", Springer-Verlag,
N.Y. (1982). Substantially pure polypeptides of at least about 90
to 95% homogeneity are preferred, and 98 to 99% or more homogeneity
are most preferred, for pharmaceutical uses. Once purified,
partially or to homogeneity as desired, the bispecific single chain
antibody molecule of the invention may then be used therapeutically
(including extracorporeally) or in developing and performing assay
procedures. Furthermore, examples for methods for the recovery of
the bispecific single chain antibody molecule of the invention from
a culture are described in detail in the appended examples. The
recovery can also be achieved by a method for the isolation of the
bispecific single chain antibody molecule of the invention capable
of binding to an epitope of human and non-chimpanzee primate CD3
epsilon (CD3.epsilon..quadrature., the method comprising the steps
of:
[0175] (a) contacting the polypeptide(s) with an N-terminal
fragment of the extracellular domain of CD3.epsilon. of maximal 27
amino acids comprising the amino acid sequence
Gln-Asp-Gly-Asn-Glu-Glu-Met-Gly (SEQ ID NO. 341) or
Gln-Asp-Gly-Asn-Glu-Glu-Ile-Gly (SEQ ID NO. 342), fixed via its
C-terminus to a solid phase;
[0176] (b) eluting the bound polypeptide(s) from said fragment;
and
[0177] (c) isolating the polypeptide(s) from the eluate of (b).
[0178] It is preferred that the polypeptide(s) isolated by the
above method of the invention are of human origin.
[0179] This method or the isolation of the bispecific single chain
antibody molecule of the invention is understood as a method for
the isolation of one or more different polypeptides with the same
specificity for the fragment of the extracellular domain of
CD3.epsilon. comprising at its N-terminus the amino acid sequence
Gln-Asp-Gly-Asn-Glu-Glu-Met-Gly (SEQ ID NO. 341) or
Gln-Asp-Gly-Asn-Glu-Glu-Ile-Gly (SEQ ID NO. 342) from a plurality
of polypeptide candidates as well as a method for the purification
of a polypeptide from a solution. A non-limiting example for the
latter method for the purification of a bispecific single chain
antibody molecule from a solution is e.g. the purification of a
recombinantly expressed bispecific single chain antibody molecule
from a culture supernatant or a preparation from such culture.
[0180] As stated above the fragment used in this method is an
N-terminal fragment of the extracellular domain of the primate
CD3.epsilon. molecule. The amino acid sequence of the extracellular
domain of the CD3.epsilon. molecule of different species is
depicted in SEQ ID NOs: 1, 3, 5 and 7. The two forms of the
N-terminal octamer are depicted in SEQ ID NOs: 341 and 342. It is
preferred that this N-terminus is freely available for binding of
the polypeptides to be identified by the method of the invention.
The term "freely available" is understood in the context of the
invention as free of additional motives such as a His-tag. The
interference of such a His-tag with a binding molecule identified
by the method of the invention is described in the appended
Examples 6 and 20.
[0181] According to this method said fragment is fixed via its
C-terminus to a solid phase. The person skilled in the art will
easily and without any inventive ado elect a suitable solid phase
support dependent from the used embodiment of the method of the
invention. Examples for a solid support comprise but are not
limited to matrices like beads (e.g. agarose beads, sepharose
beads, polystyrol beads, dextran beads), plates (culture plates or
MultiWell plates) as well as chips known e.g. from Biacore.RTM..
The selection of the means and methods for the
fixation/immobilization of the fragment to said solid support
depend on the election of the solid support. A commonly used method
for the fixation/immobilization is a coupling via an
N-hydroxysuccinimide (NHS) ester. The chemistry underlying this
coupling as well as alternative methods for the
fixation/immobilization are known to the person skilled in the art,
e.g. from Hermanson "Bioconjugate Techniques", Academic Press, Inc.
(1996). For the fixation to/immobilization on chromatographic
supports the following means are commonly used: NHS-activated
sepharose (e.g. HiTrap-NHS of GE Life Science-Amersham),
CnBr-activated sepharose (e.g. GE Life Science-Amersham),
NHS-activated dextran beads (Sigma) or activated polymethacrylate.
These reagents may also be used in a batch approach. Moreover,
dextran beads comprising iron oxide (e.g. available from Miltenyi)
may be used in a batch approach. These beads may be used in
combination with a magnet for the separation of the beads from a
solution. Polypeptides can be immobilized on a Biacore chip (e.g.
CM5 chips) by the use of NHS activated carboxymethyldextran.
Further examples for an appropriate solid support are amine
reactive MultiWell plates (e.g. Nunc Immobilizer.TM. plates).
[0182] According to this method said fragment of the extracellular
domain of CD3 epsilon can be directly coupled to the solid support
or via a stretch of amino acids, which might be a linker or another
protein/polypeptide moiety. Alternatively, the extracellular domain
of CD3 epsilon can be indirectly coupled via one or more adaptor
molecule(s).
[0183] Means and methods for the eluation of a peptide or
polypeptide bound to an immobilized epitope are well known in the
art. The same holds true for methods for the isolation of the
identified polypeptide(s) from the eluate.
[0184] A method for the isolation of one or more different
bispecific single chain antibody molecule(s) with the same
specificity for the fragment of the extracellular domain of
CD3.epsilon. comprising at its N-terminus the amino acid sequence
Gln-Asp-Gly-Asn-Glu-Glu-X-Gly (with X being Met or Ile) from a
plurality of polypeptide candidates may comprise one or more steps
of the following methods for the selection of antigen-specific
entities:
[0185] CD3.epsilon. specific binding domains can be selected from
antibody derived repertoires. A phage display library can be
constructed based on standard procedures, as for example disclosed
in "Phage Display: A Laboratory Manual"; Ed. Barbas, Burton, Scott
& Silverman; Cold Spring Harbor Laboratory Press, 2001. The
format of the antibody fragments in the antibody library can be
scFv, but may generally also be a Fab fragment or even a single
domain antibody fragment. For the isolation of antibody fragments
naive antibody fragment libraries may be used. For the selection of
potentially low immunogenic binding entities in later therapeutic
use, human antibody fragment libraries may be favourable for the
direct selection of human antibody fragments. In some cases they
may form the basis for synthetic antibody libraries (Knappik et al.
J Mol. Biol. 2000, 296:57 ff). The corresponding format may be Fab,
scFv (as described below) or domain antibodies (dAbs, as reviewed
in Holt et al., Trends Biotechnol. 2003, 21:484 ff).
[0186] It is also known in the art that in many cases there is no
immune human antibody source available against the target antigen.
Therefore animals are immunized with the target antigen and the
respective antibody libraries isolated from animal tissue as e.g.
spleen or PBMCs. The N-terminal fragment may be biotinylated or
covalently linked to proteins like KLH or bovine serum albumin
(BSA). According to common approaches rodents are used for
immunization. Some immune antibody repertoires of non-human origin
may be especially favourable for other reasons, e.g. for the
presence of single domain antibodies (VHH) derived from cameloid
species (as described in Muyldermans, J Biotechnol. 74:277; De
Genst et al. Dev Como Immunol. 2006, 30:187 ff). Therefore a
corresponding format of the antibody library may be Fab, scFv (as
described below) or single domain antibodies (VHH).
[0187] In one possible approach ten weeks old F1 mice from
balb/c.times.C57black crossings can be immunized with whole cells
e.g. expressing transmembrane EpCAM N-terminally displaying as
translational fusion the N-terminal amino acids 1 to 27 of the
mature CD3.epsilon. chain. Alternatively, mice can be immunized
with 1-27 CD3 epsilon-Fc fusion protein (a corresponding approach
is described in the appended Example 2). After booster
immunization(s), blood samples can be taken and antibody serum
titer against the CD3-positive T cells can be tested e.g. in FACS
analysis. Usually, serum titers are significantly higher in
immunized than in non-immunized animals.
[0188] Immunized animals may form the basis for the construction of
immune antibody libraries. Examples of such libraries comprise
phage display libraries. Such libraries may be generally
constructed based on standard procedures, as for example disclosed
in "Phage Display: A Laboratory Manual"; Ed. Barbas, Burton, Scott
& Silverman; Cold Spring Harbor Laboratory Press, 2001.
[0189] The non-human antibodies can also be humanized via phage
display due to the generation of more variable antibody libraries
that can be subsequently enriched for binders during selection.
[0190] In a phage display approach any one of the pools of phages
that displays the antibody libraries forms a basis to select
binding entities using the respective antigen as target molecule.
The central step in which antigen specific, antigen bound phages
are isolated is designated as panning. Due to the display of the
antibody fragments on the surface of the phages, this general
method is called phage display. One preferred method of selection
is the use of small proteins such as the filamentous phage N2
domain translationally fused to the N-terminus of the scFv
displayed by the phage. Another display method known in the art,
which may be used to isolate binding entities is the ribosome
display method (reviewed in Groves & Osbourn, Expert Opin Biol
Ther. 2005, 5:125 ff; Lipovsek & Pluckthun, J Immunol Methods
2004, 290:52 ff). In order to demonstrate binding of scFv phage
particles to a 1-27 CD3.epsilon.-Fc fusion protein a phage library
carrying the cloned scFv-repertoire can be harvested from the
respective culture supernatant by PEG (polyethyleneglycole). ScFv
phage particles may be incubated with immobilized CD3.epsilon. Fc
fusion protein. The immobilized CD3.epsilon. Fc fusion protein may
be coated to a solid phase. Binding entities can be eluted and the
eluate can be used for infection of fresh uninfected bacterial
hosts. Bacterial hosts successfully transduced with a phagemid
copy, encoding a human scFv-fragment, can be selected again for
carbenicillin resistance and subsequently infected with e.g. VCMS
13 helper phage to start the second round of antibody display and
in vitro selection. A total of 4 to 5 rounds of selections is
carried out, normally. The binding of isolated binding entities can
be tested on CD3 epsilon positive Jurkat cells, HPBall cells, PBMCs
or transfected eukaryotic cells that carry the N-terminal
CD3.epsilon. sequence fused to surface displayed EpCAM using a flow
cytometric assay (see appended Example 4).
[0191] Preferably, the above method may be a method, wherein the
fragment of the extracellular domain of CD3.epsilon. consists of
one or more fragments of a polypeptide having an amino acid
sequence of any one depicted in SEQ ID NOs. 2, 4, 6 or 8. More
preferably, said fragment is 8, 9, 10, 11, 12, 13, 14, 15, 16, 17,
18, 19, 20, 21, 22, 23, 24, 25, 26, 27 amino acid residues in
length.
[0192] This method of identification of a bispecific single chain
antibody molecule may be a method of screening a plurality of
bispecific single chain antibody molecules comprising a
cross-species specific binding domain binding to an epitope of
human and non-chimpanzee primate CD3.epsilon.. Alternatively, the
method of identification is a method of purification/isolation of a
bispecific single chain antibody molecule comprising a
cross-species specific binding domain binding to an epitope of
human and non-chimpanzee primate CD3.epsilon..
[0193] Furthermore, the invention provides for a composition
comprising a bispecific single chain antibody molecule of the
invention or a bispecific single chain antibody as produced by the
process disclosed above. Preferably, said composition is a
pharmaceutical composition.
[0194] The invention provides also for a bispecific single chain
antibody molecule as defined herein, or produced according to the
process as defined herein, wherein said bispecific single chain
antibody molecule is for use in the prevention, treatment or
amelioration of cancer. Preferably, said cancer is a solid tumor,
more preferably a carcinoma or prostate cancer. It is preferred
that the bispecific single chain is further comprising suitable
formulations of carriers, stabilizers and/or excipients. Moreover,
it is preferred that said bispecific single chain antibody molecule
is suitable to be administered in combination with an additional
drug. Said drug may be a non-proteinaceous compound or a
proteinaceous compound and may be administered simultaneously or
non-simultaneously with the bispecific single chain antibody
molecule as defined herein.
[0195] In accordance with the invention, the term "pharmaceutical
composition" relates to a composition for administration to a
patient, preferably a human patient. The particular preferred
pharmaceutical composition of this invention comprises bispecific
single chain antibodies directed against and generated against
context-independent CD3 epitopes. Preferably, the pharmaceutical
composition comprises suitable formulations of carriers,
stabilizers and/or excipients. In a preferred embodiment, the
pharmaceutical composition comprises a composition for parenteral,
transdermal, intraluminal, intraarterial, intrathecal and/or
intranasal administration or by direct injection into tissue. It is
in particular envisaged that said composition is administered to a
patient via infusion or injection. Administration of the suitable
compositions may be effected by different ways, e.g., by
intravenous, intraperitoneal, subcutaneous, intramuscular, topical
or intradermal administration. In particular, the present invention
provides for an uninterrupted administration of the suitable
composition. As a non-limiting example, uninterrupted, i.e.
continuous administration may be realized by a small pump system
worn by the patient for metering the influx of therapeutic agent
into the body of the patient. The pharmaceutical composition
comprising the bispecific single chain antibodies directed against
and generated against context-independent CD3 epitopes of the
invention can be administered by using said pump systems. Such pump
systems are generally known in the art, and commonly rely on
periodic exchange of cartridges containing the therapeutic agent to
be infused. When exchanging the cartridge in such a pump system, a
temporary interruption of the otherwise uninterrupted flow of
therapeutic agent into the body of the patient may ensue. In such a
case, the phase of administration prior to cartridge replacement
and the phase of administration following cartridge replacement
would still be considered within the meaning of the pharmaceutical
means and methods of the invention together make up one
"uninterrupted administration" of such therapeutic agent.
[0196] The continuous or uninterrupted administration of these
bispecific single chain antibodies directed against and generated
against context-independent CD3 epitopes of this invention may be
intravenuous or subcutaneous by way of a fluid delivery device or
small pump system including a fluid driving mechanism for driving
fluid out of a reservoir and an actuating mechanism for actuating
the driving mechanism. Pump systems for subcutaneous administration
may include a needle or a cannula for penetrating the skin of a
patient and delivering the suitable composition into the patient's
body. Said pump systems may be directly fixed or attached to the
skin of the patient independently of a vein, artery or blood
vessel, thereby allowing a direct contact between the pump system
and the skin of the patient. The pump system can be attached to the
skin of the patient for 24 hours up to several days. The pump
system may be of small size with a reservoir for small volumes. As
a non-limiting example, the volume of the reservoir for the
suitable pharmaceutical composition to be administered can be
between 0.1 and 50 ml.
[0197] The continuous administration may be transdermal by way of a
patch worn on the skin and replaced at intervals. One of skill in
the art is aware of patch systems for drug delivery suitable for
this purpose. It is of note that transdermal administration is
especially amenable to uninterrupted administration, as exchange of
a first exhausted patch can advantageously be accomplished
simultaneously with the placement of a new, second patch, for
example on the surface of the skin immediately adjacent to the
first exhausted patch and immediately prior to removal of the first
exhausted patch. Issues of flow interruption or power cell failure
do not arise.
[0198] The composition of the present invention, comprising in
particular bispecific single chain antibodies directed against and
generated against context-independent CD3 epitopes may further
comprise a pharmaceutically acceptable carrier. Examples of
suitable pharmaceutical carriers are well known in the art and
include solutions, e.g. phosphate buffered saline solutions, water,
emulsions, such as oil/water emulsions, various types of wetting
agents, sterile solutions, liposomes, etc. Compositions comprising
such carriers can be formulated by well known conventional methods.
Formulations can comprise carbohydrates, buffer solutions, amino
acids and/or surfactants. Carbohydrates may be non-reducing sugars,
preferably trehalose, sucrose, octasulfate, sorbitol or xylitol.
Such formulations may be used for continuous administrations which
may be intravenuous or subcutaneous with and/or without pump
systems. Amino acids may be charged amino acids, preferably lysine,
lysine acetate, arginine, glutamate and/or histidine. Surfactants
may be detergents, preferably with a molecular weight of >1.2 KD
and/or a polyether, preferably with a molecular weight of >3 KD.
Non-limiting examples for preferred detergents are Tween 20, Tween
40, Tween 60, Tween 80 or Tween 85. Non-limiting examples for
preferred polyethers are PEG 3000, PEG 3350, PEG 4000 or PEG 5000.
Buffer systems used in the present invention can have a preferred
pH of 5-9 and may comprise citrate, succinate, phosphate, histidine
and acetate. The compositions of the present invention can be
administered to the subject at a suitable dose which can be
determined e.g. by dose escalating studies by administration of
increasing doses of the bispecific single chain antibody molecule
of the invention exhibiting cross-species specificity described
herein to non-chimpanzee primates, for instance macaques. As set
forth above, the bispecific single chain antibody molecule of the
invention exhibiting cross-species specificity described herein can
be advantageously used in identical form in preclinical testing in
non-chimpanzee primates and as drug in humans. These compositions
can also be administered in combination with other proteinaceous
and non-proteinaceous drugs. These drugs may be administered
simultaneously with the composition comprising the bispecific
single chain antibody molecule of the invention as defined herein
or separately before or after administration of said polypeptide in
timely defined intervals and doses. The dosage regimen will be
determined by the attending physician and clinical factors. As is
well known in the medical arts, dosages for any one patient depend
upon many factors, including the patient's size, body surface area,
age, the particular compound to be administered, sex, time and
route of administration, general health, and other drugs being
administered concurrently. Preparations for parenteral
administration include sterile aqueous or non-aqueous solutions,
suspensions, and emulsions. Examples of non-aqueous solvents are
propylene glycol, polyethylene glycol, vegetable oils such as olive
oil, and injectable organic esters such as ethyl oleate. Aqueous
carriers include water, alcoholic/aqueous solutions, emulsions or
suspensions, including saline and buffered media. Parenteral
vehicles include sodium chloride solution, Ringer's dextrose,
dextrose and sodium chloride, lactated Ringer's, or fixed oils.
[0199] Intravenous vehicles include fluid and nutrient
replenishers, electrolyte replenishers (such as those based on
Ringer's dextrose), and the like. Preservatives and other additives
may also be present such as, for example, antimicrobials,
anti-oxidants, chelating agents, inert gases and the like. In
addition, the composition of the present invention might comprise
proteinaceous carriers, like, e.g., serum albumin or
immunoglobulin, preferably of human origin. It is envisaged that
the composition of the invention might comprise, in addition to the
bispecific single chain antibody molecule of the invention defined
herein, further biologically active agents, depending on the
intended use of the composition. Such agents might be drugs acting
on the gastro-intestinal system, drugs acting as cytostatica, drugs
preventing hyperurikemia, drugs inhibiting immunoreactions (e.g.
corticosteroids), drugs modulating the inflammatory response, drugs
acting on the circulatory system and/or agents such as cytokines
known in the art.
[0200] The biological activity of the pharmaceutical composition
defined herein can be determined for instance by cytotoxicity
assays, as described in the following examples, in WO 99/54440 or
by Schlereth et al. (Cancer Immunol. Immunother. 20 (2005), 1-12).
"Efficacy" or "in vivo efficacy" as used herein refers to the
response to therapy by the pharmaceutical composition of the
invention, using e.g. standardized NCI response criteria. The
success or in vivo efficacy of the therapy using a pharmaceutical
composition of the invention refers to the effectiveness of the
composition for its intended purpose, i.e. the ability of the
composition to cause its desired effect, i.e. depletion of
pathologic cells, e.g. tumor cells. The in vivo efficacy may be
monitored by established standard methods for the respective
disease entities including, but not limited to white blood cell
counts, differentials, Fluorescence Activated Cell Sorting, bone
marrow aspiration. In addition, various disease specific clinical
chemistry parameters and other established standard methods may be
used. Furthermore, computer-aided tomography, X-ray, nuclear
magnetic resonance tomography (e.g. for National Cancer
Institute-criteria based response assessment [Cheson B D, Horning S
J, Coiffier B, Shipp M A, Fisher R I, Connors J M, Lister T A, Vose
J, Grillo-Lopez A, Hagenbeek A, Cabanillas F, Klippensten D,
Hiddemann W, Castellino R, Harris N L, Armitage J O, Carter W,
Hoppe R, Canellos G P. Report of an international workshop to
standardize response criteria for non-Hodgkin's lymphomas. NCI
Sponsored International Working Group. J Clin Oncol. 1999 April;
17(4):1244]), positron-emission tomography scanning, white blood
cell counts, differentials, Fluorescence Activated Cell Sorting,
bone marrow aspiration, lymph node biopsies/histologies, and
various cancer specific clinical chemistry parameters (e.g. lactate
dehydrogenase) and other established standard methods may be
used.
[0201] Another major challenge in the development of drugs such as
the pharmaceutical composition of the invention is the predictable
modulation of pharmacokinetic properties. To this end, a
pharmacokinetic profile of the drug candidate, i.e. a profile of
the pharmacokinetic parameters that effect the ability of a
particular drug to treat a given condition, is established.
Pharmacokinetic parameters of the drug influencing the ability of a
drug for treating a certain disease entity include, but are not
limited to: half-life, volume of distribution, hepatic first-pass
metabolism and the degree of blood serum binding. The efficacy of a
given drug agent can be influenced by each of the parameters
mentioned above.
[0202] "Half-life" means the time where 50% of an administered drug
are eliminated through biological processes, e.g. metabolism,
excretion, etc.
[0203] By "hepatic first-pass metabolism" is meant the propensity
of a drug to be metabolized upon first contact with the liver, i.e.
during its first pass through the liver.
[0204] "Volume of distribution" means the degree of retention of a
drug throughout the various compartments of the body, like e.g.
intracellular and extracellular spaces, tissues and organs, etc.
and the distribution of the drug within these compartments.
[0205] "Degree of blood serum binding" means the propensity of a
drug to interact with and bind to blood serum proteins, such as
albumin, leading to a reduction or loss of biological activity of
the drug.
[0206] Pharmacokinetic parameters also include bioavailability, lag
time (Tlag), Tmax, absorption rates, more onset and/or Cmax for a
given amount of drug administered.
[0207] "Bioavailability" means the amount of a drug in the blood
compartment.
[0208] "Lag time" means the time delay between the administration
of the drug and its detection and measurability in blood or
plasma.
[0209] "Tmax" is the time after which maximal blood concentration
of the drug is reached, and "Cmax" is the blood concentration
maximally obtained with a given drug. The time to reach a blood or
tissue concentration of the drug which is required for its
biological effect is influenced by all parameters. Pharmacokinetik
parameters of bispecific single chain antibodies exhibiting
cross-species specificity, which may be determined in preclinical
animal testing in non-chimpanzee primates as outlined above are
also set forth e.g. in the publication by Schlereth et al. (Cancer
Immunol. Immunother. 20 (2005), 1-12).
[0210] The term "toxicity" as used herein refers to the toxic
effects of a drug manifested in adverse events or severe adverse
events. These side events might refer to a lack of tolerability of
the drug in general and/or a lack of local tolerance after
administration. Toxicity could also include teratogenic or
carcinogenic effects caused by the drug.
[0211] The term "safety", "in vivo safety" or "tolerability" as
used herein defines the administration of a drug without inducing
severe adverse events directly after administration (local
tolerance) and during a longer period of application of the drug.
"Safety", "in vivo safety" or "tolerability" can be evaluated e.g.
at regular intervals during the treatment and follow-up period.
Measurements include clinical evaluation, e.g. organ
manifestations, and screening of laboratory abnormalities. Clinical
evaluation may be carried out and deviating to normal findings
recorded/coded according to NCI-CTC and/or MedDRA standards. Organ
manifestations may include criteria such as allergy/immunology,
blood/bone marrow, cardiac arrhythmia, coagulation and the like, as
set forth e.g. in the Common Terminology Criteria for adverse
events v3.0 (CTCAE). Laboratory parameters which may be tested
include for instance haematology, clinical chemistry, coagulation
profile and urine analysis and examination of other body fluids
such as serum, plasma, lymphoid or spinal fluid, liquor and the
like. Safety can thus be assessed e.g. by physical examination,
imaging techniques (i.e. ultrasound, x-ray, CT scans, Magnetic
Resonance Imaging (MRI), other measures with technical devices
(i.e. electrocardiogram), vital signs, by measuring laboratory
parameters and recording adverse events. For example, adverse
events in non-chimpanzee primates in the uses and methods according
to the invention may be examined by histopathological and/or
histochemical methods.
[0212] The term "effective and non-toxic dose" as used herein
refers to a tolerable dose of the bispecific single chain antibody
as defined herein which is high enough to cause depletion of
pathologic cells, tumor elimination, tumor shrinkage or
stabilization of disease without or essentially without major toxic
effects. Such effective and non-toxic doses may be determined e.g.
by dose escalation studies described in the art and should be below
the dose inducing severe adverse side events (dose limiting
toxicity, DLT).
[0213] The above terms are also referred to e.g. in the Preclinical
safety evaluation of biotechnology-derived pharmaceuticals S6; ICH
Harmonised Tripartite Guideline; ICH Steering Committee meeting on
Jul. 16, 1997.
[0214] Moreover, the invention relates to a pharmaceutical
composition comprising a bispecific single chain antibody molecule
of this invention or produced according to the process according to
the invention for the prevention, treatment or amelioration of
cancer. Preferably, said cancer is a solid tumor, preferably a
carcinoma or prostate cancer. Preferably, said pharmaceutical
composition further comprises suitable formulations of carriers,
stabilizers and/or excipients.
[0215] A further aspect of the invention relates to a use of a
bispecific single chain antibody molecule/polypeptide as defined
herein above or produced according to a process defined herein
above, for the preparation of a pharmaceutical composition for the
prevention, treatment or amelioration of a disease. Preferably,
said disease is cancer. More preferably, said cancer is a solid
tumor, preferably a carcinoma or prostate cancer.
[0216] In another preferred embodiment of use of the bispecific
single chain antibody molecule of the invention said pharmaceutical
composition is suitable to be administered in combination with an
additional drug, i.e. as part of a co-therapy. In said co-therapy,
an active agent may be optionally included in the same
pharmaceutical composition as the bispecific single chain antibody
molecule of the invention, or may be included in a separate
pharmaceutical composition. In this latter case, said separate
pharmaceutical composition is suitable for administration prior to,
simultaneously as or following administration of said
pharmaceutical composition comprising the bispecific single chain
antibody molecule of the invention. The additional drug or
pharmaceutical composition may be a non-proteinaceous compound or a
proteinaceous compound. In the case that the additional drug is a
proteinaceous compound, it is advantageous that the proteinaceous
compound be capable of providing an activation signal for immune
effector cells.
[0217] Preferably, said proteinaceous compound or non-proteinaceous
compound may be administered simultaneously or non-simultaneously
with the bispecific single chain antibody molecule of the
invention, a nucleic acid molecule as defined hereinabove, a vector
as defined as defined hereinabove, or a host as defined as defined
hereinabove.
[0218] Another aspect of the invention relates to a method for the
prevention, treatment or amelioration of a disease in a subject in
the need thereof, said method comprising the step of administration
of an effective amount of a pharmaceutical composition of the
invention. Preferably, said disease is cancer. Preferably, said
cancer is a solid tumor, preferably a carcinoma or prostate
cancer.
[0219] In another preferred embodiment of the method of the
invention said pharmaceutical composition is suitable to be
administered in combination with an additional drug, i.e. as part
of a co-therapy. In said co-therapy, an active agent may be
optionally included in the same pharmaceutical composition as the
bispecific single chain antibody molecule of the invention, or may
be included in a separate pharmaceutical composition. In this
latter case, said separate pharmaceutical composition is suitable
for administration prior to, simultaneously as or following
administration of said pharmaceutical composition comprising the
bispecific single chain antibody molecule of the invention. The
additional drug or pharmaceutical composition may be a
non-proteinaceous compound or a proteinaceous compound. In the case
that the additional drug is a proteinaceous compound, it is
advantageous that the proteinaceous compound be capable of
providing an activation signal for immune effector cells.
[0220] Preferably, said proteinaceous compound or non-proteinaceous
compound may be administered simultaneously or non-simultaneously
with the bispecific single chain antibody molecule of the
invention, a nucleic acid molecule as defined hereinabove, a vector
as defined as defined hereinabove, or a host as defined as defined
hereinabove.
[0221] It is preferred for the above described method of the
invention that said subject is a human.
[0222] In a further aspect, the invention relates to a kit
comprising a bispecific single chain antibody molecule of the
invention, a nucleic acid molecule of the invention, a vector of
the invention, or a host of the invention.
[0223] These and other embodiments are disclosed and encompassed by
the description and Examples of the present invention. Recombinant
techniques and methods in immunology are described e.g. in Sambrook
et al. Molecular Cloning: A Laboratory Manual; Cold Spring Harbor
Laboratory Press, 3.sup.rd edition 2001; Lefkovits; Immunology
Methods Manual; The Comprehensive Sourcebook of Techniques;
Academic Press, 1997; Golemis; Protein-Protein Interactions: A
Molecular Cloning Manual; Cold Spring Laboratory Press, 2002.
Further literature concerning any one of the antibodies, methods,
uses and compounds to be employed in accordance with the present
invention may be retrieved from public libraries and databases,
using for example electronic devices.--
[0224] The figures show:
[0225] FIG. 1
[0226] Fusion of the N-terminal amino acids 1-27 of primate CD3
epsilon to a heterologous soluble protein.
[0227] FIG. 2
[0228] The figure shows the average absorption values of
quadruplicate samples measured in an ELISA assay detecting the
presence of a construct consisting of the N-terminal amino acids
1-27 of the mature human CD3 epsilon chain fused to the hinge and
Fc gamma portion of human IgG1 and a C-terminal 6 Histidine tag in
a supernatant of transiently transfected 293 cells. The first
column labeled "27 aa huCD3E" shows the average absorption value
for the construct, the second column labeled "irrel. SN" shows the
average value for a supernatant of 293 cells transfected with an
irrelevant construct as negative control. The comparison of the
values obtained for the construct with the values obtained for the
negative control clearly demonstrates the presence of the
recombinant construct.
[0229] FIG. 3
[0230] The figure shows the average absorption values of
quadruplicate samples measured in an ELISA assay detecting the
binding of the cross species specific anti-CD3 binding molecules in
form of crude preparations of periplasmatically expressed
single-chain antibodies to a construct comprising the N-terminal
1-27 amino acids of the mature human CD3 epsilon chain fused to the
hinge and Fc gamma portion of human IgG1 and a C-terminal His6 tag.
The columns show from left to right the average absorption values
for the specificities designated as A2J HLP, I2C HLP E2M HLP, F7O
HLP, G4H HLP, H2C HLP, E1L HLP, F12Q HLP, F6A HLP and H1E HLP. The
rightmost column labelled "neg. contr." shows the average
absorption value for the single-chain preparation of a murine
anti-human CD3 antibody as negative control. The comparison of the
values obtained for the anti-CD3 specificities with the values
obtained for the negative control clearly demonstrates the strong
binding of the anti-CD3 specificities to the N-terminal 1-27 amino
acids of the mature human CD3 epsilon chain.
[0231] FIG. 4
[0232] Fusion of the N-terminal amino acids 1-27 of primate CD3
epsilon to a heterologous membrane bound protein.
[0233] FIG. 5
[0234] Histogram overlays of different transfectants tested in a
FACS assay detecting the presence of recombinant transmembrane
fusion proteins consisting of cynomolgus EpCAM and the N-terminal
1-27 amino acids of the human, marmoset, tamarin, squirrel monkey
and domestic swine CD3 epsilon chain respectively. The histogram
overlays from left to right and top to bottom show the results for
the transfectants expressing the constructs comprising the human 27
mer, marmoset 27 mer, tamarin 27 mer, squirrel monkey 27 mer and
swine 27 mer respectively. In the individual overlays the thin line
represents a sample incubated with PBS with 2% FCS instead of
anti-Flag M2 antibody as negative control and the bold line shows a
sample incubated with the anti-Flag M2 antibody. For each construct
the overlay of the histograms shows binding of the anti-Flag M2
antibody to the transfectants, which clearly demonstrates the
expression of the recombinant constructs on the transfectants.
[0235] FIGS. 6A-6E
[0236] Histogram overlays of different transfectants tested in a
FACS assay detecting the binding of the cross-species specific
anti-CD3 binding molecules in form of crude preparations of
periplasmatically expressed single-chain antibodies to the
N-terminal amino acids 1-27 of the human, marmoset, tamarin and
squirrel monkey CD3 epsilon chain respectively fused to cynomolgus
EpCAM.
[0237] FIG. 6A:
[0238] The histogram overlays from left to right and top to bottom
show the results for the transfectants expressing the 1-27
CD3-EpCAM comprising the human 27 mer tested with the CD3 specific
binding molecules designated H2C HLP, F12Q HLP, E2M HLP and G4H HLP
respectively.
[0239] FIG. 6B:
[0240] The histogram overlays from left to right and top to bottom
show the results for the transfectants expressing the 1-27
CD3-EpCAM comprising the marmoset 27 mer tested with the CD3
specific binding molecules designated H2C HLP, F12Q HLP, E2M HLP
and G4H HLP respectively.
[0241] FIG. 6C:
[0242] The histogram overlays from left to right and top to bottom
show the results for the transfectants expressing the 1-27
CD3-EpCAM comprising the tamarin 27 mer tested with the CD3
specific binding molecules designated H2C HLP, F12Q HLP, E2M HLP
and G4H HLP respectively.
[0243] FIG. 6D:
[0244] The histogram overlays from left to right and top to bottom
show the results for the transfectants expressing the 1-27
CD3-EpCAM comprising the squirrel monkey 27 mer tested with the CD3
specific binding molecules designated H2C HLP, F12Q HLP, E2M HLP
and G4H HLP respectively.
[0245] FIG. 6E:
[0246] The histogram overlays from left to right and top to bottom
show the results for the transfectants expressing the 1-27
CD3-EpCAM comprising the swine 27 mer tested with the CD3 specific
binding molecules designated H2C HLP, F12Q HLP, E2M HLP and G4H HLP
respectively.
[0247] In the individual overlays the thin line represents a sample
incubated with a single-chain preparation of a murine anti-human
CD3-antibody as negative control and the bold line shows a sample
incubated with the respective anti-CD3 binding molecules indicated.
Considering the lack of binding to the swine 27 mer transfectants
and the expression levels of the constructs shown in FIG. 5 the
overlays of the histograms show specific and strong binding of the
tested anti-CD3 specificities of the fully cross-species specific
human bispecific single chain antibodies to cells expressing the
recombinant transmembrane fusion proteins comprising the N-terminal
amino acids 1-27 of the human, marmoset, tamarin and squirrel
monkey CD3 epsilon chain respectively fused to cynomolgus EpCAM and
show therefore multi primate cross-species specificity of the
anti-CD3 binding molecules.
[0248] FIG. 7
[0249] FACS assay for detection of human CD3 epsilon on transfected
murine EL4 T cells. Graphical analysis shows an overlay of
histograms. The bold line shows transfected cells incubated with
the anti-human CD3 antibody UCHT-1. The thin line represents cells
incubated with a mouse IgG1 isotype control. Binding of the anti
CD3 antibody UCHT1 clearly shows expression of the human CD3
epsilon chain on the cell surface of transfected murine EL4 T
cells.
[0250] FIGS. 8A-8D
[0251] Binding of cross-species specific anti CD3 antibodies to
alanine-mutants in an alanine scanning experiment. In the
individual Figures the columns show from left to right the
calculated binding values in arbitrary units in logarithmic scale
for the wild-type transfectant (WT) and for all alanine-mutants
from the position 1 to 27. The binding values are calculated using
the following formula:
value_Sample ( x , y ) = Sample ( x , y ) - neg_Contr . ( x ) (
UCHT - 1 ( x ) - neg_Contr . ( x ) ) * WT ( y ) - neg_Contr . ( wt
) UCHT - 1 ( wt ) - neg_Contr . ( wt ) ##EQU00001##
[0252] In this equation value_Sample means the value in arbitrary
units of binding depicting the degree of binding of a specific
anti-CD3 antibody to a specific alanine-mutant as shown in the
Figure, Sample means the geometric mean fluorescence value obtained
for a specific anti-CD3 antibody assayed on a specific
alanine-scanning transfectant, neg_Contr. means the geometric mean
fluorescence value obtained for the negative control assayed on a
specific alanine-mutant, UCHT-1 means the geometric mean
fluorescence value obtained for the UCHT-1 antibody assayed on a
specific alanine-mutant, WT means the geometric mean fluorescence
value obtained for a specific anti-CD3 antibody assayed on the
wild-type transfectant, x specifies the respective transfectant, y
specifies the respective anti-CD3 antibody and wt specifies that
the respective transfectant is the wild-type. Individual
alanine-mutant positions are labelled with the single letter code
of the wild-type amino acid and the number of the position.
[0253] FIG. 8A:
[0254] The figure shows the results for cross-species specific anti
CD3 antibody A2J HLP expressed as chimeric IgG molecule. Reduced
binding activity is observed for mutations to alanine at position 4
(asparagine), at position 23 (threonine) and at position 25
(isoleucine). Complete loss of binding is observed for mutations to
alanine at position 1 (glutamine), at position 2 (aspartate), at
position 3 (glycine) and at position 5 (glutamate).
[0255] FIG. 8B:
[0256] The figure shows the results for cross-species specific anti
CD3 antibody E2M HLP, expressed as chimeric IgG molecule. Reduced
binding activity is observed for mutations to alanine at position 4
(asparagine), at position 23 (threonine) and at position 25
(isoleucine). Complete loss of binding is observed for mutations to
alanine at position 1 (glutamine), at position 2 (aspartate), at
position 3 (glycine) and at position 5 (glutamate).
[0257] FIG. 8C:
[0258] The figure shows the results for cross-species specific anti
CD3 antibody H2C HLP, expressed as chimeric IgG molecule. Reduced
binding activity is observed for mutations to alanine at position 4
(asparagine). Complete loss of binding is observed for mutations to
alanine glutamine at position 1 (glutamine), at position 2
(aspartate), at position 3 (glycine) and at position 5
(glutamate).
[0259] FIG. 8D:
[0260] shows the results for cross-species specific anti CD3
antibody F12Q HLP, tested as periplasmatically expressed
single-chain antibody. Complete loss of binding is observed for
mutations to alanine at position 1 (glutamine), at position 2
(aspartate), at position 3 (glycine) and at position 5
(glutamate).
[0261] FIG. 9
[0262] FACS assay detecting the binding of the cross-species
specific anti-CD3 binding molecule H2C HLP to human CD3 with and
without N-terminal His6 tag.
[0263] Histogram overlays are performed of the EL4 cell line
transfected with wild-type human CD3 epsilon chain (left histogram)
or the human CD3 epsilon chain with N-terminal His6 tag (right
histogram) tested in a FACS assay detecting the binding of
cross-species specific binding molecule H2C HLP. Samples are
incubated with an appropriate isotype control as negative control
(thin line), anti-human CD3 antibody UCHT-1 as positive control
(dotted line) and cross-species specific anti-CD3 antibody H2C HLP
in form of a chimeric IgG molecule (bold line).
[0264] Histogram overlays show comparable binding of the UCHT-1
antibody to both transfectants as compared to the isotype control
demonstrating expression of both recombinant constructs. Histogram
overlays also show binding of the anti-CD3 binding molecule H2C HLP
only to the wild-type human CD3 epsilon chain but not to the
His6-human CD3 epsilon chain. These results demonstrate that a free
N-terminus is essential for binding of the cross-species specific
anti-CD3 binding molecule H2C HLP.
[0265] FIG. 10
[0266] FACS binding analysis of designated cross-species specific
bispecific single chain constructs to CHO cells transfected with
the human MCSP D3, human CD3+ T cell line HPB-ALL, CHO cells
transfected with cynomolgus MCSP D3 and a macaque T cell line 4119
LnPx. The FACS staining is performed as described in Example 10.
The thick line represents cells incubated with 2 .mu.g/ml purified
protein that are subsequently incubated with the anti-his antibody
and the PE labeled detection antibody. The thin histogram line
reflects the negative control: cells only incubated with the
anti-his antibody and the detection antibody.
[0267] FIG. 11
[0268] FACS binding analysis of designated cross-species specific
bispecific single chain constructs CHO cells transfected with the
human MCSP D3, human CD3+ T cell line HPB-ALL, CHO cells
transfected with cynomolgus MCSP D3 and a macaque T cell line 4119
LnPx. The FACS staining is performed as described in Example 10.
The thick line represents cells incubated with 2 .mu.g/ml purified
protein that are subsequently incubated with the anti-his antibody
and the PE labeled detection antibody. The thin histogram line
reflects the negative control: cells only incubated with the
anti-his antibody and the detection antibody.
[0269] FIG. 12
[0270] FACS binding analysis of designated cross-species specific
bispecific single chain constructs CHO cells transfected with the
human MCSP D3, human CD3+ T cell line HPB-ALL, CHO cells
transfected with cynomolgus MCSP D3 and a macaque T cell line 4119
LnPx. The FACS staining is performed as described in Example 10.
The thick line represents cells incubated with 2 .mu.g/ml purified
monomeric protein that are subsequently incubated with the anti-his
antibody and the PE labeled detection antibody. The thin histogram
line reflects the negative control: cells only incubated with the
anti-his antibody and the detection antibody.
[0271] FIGS. 13A-13B
[0272] Cytotoxicity activity induced by designated cross-species
specific MCSP specific single chain constructs redirected to
indicated target cell lines. FIG. 13A) Stimulated CD4-/CD56- human
PBMCs are used as effector cells, CHO cells transfected with human
MCSP D3 as target cells. FIG. 13B) The macaque T cell line 4119
LnPx are used as effector cells, CHO cells transfected with
cynomolgus MCSP D3 as target cells. The assay is performed as
described in Example 11.
[0273] FIGS. 14A-14B
[0274] Cytotoxicity activity induced by designated cross-species
specific MCSP specific single chain constructs redirected to
indicated target cell lines. FIG. 14A) and FIG. 14B) The macaque T
cell line 4119 LnPx are used as effector cells, CHO cells
transfected with cynomolgus MCSP D3 as target cells. The assay is
performed as described in Example 11.
[0275] FIGS. 15A-15B
[0276] Cytotoxicity activity induced by designated cross-species
specific MCSP specific single chain constructs redirected to
indicated target cell lines. FIG. 15A) and FIG. 15B) Stimulated
CD4-/CD56- human PBMCs are used as effector cells, CHO cells
transfected with human MCSP D3 as target cells. The assay is
performed as described in Example 11.
[0277] FIGS. 16A-16B
[0278] Cytotoxicity activity induced by designated cross-species
specific MCSP specific single chain constructs redirected to
indicated target cell lines. FIG. 16A) Stimulated CD4-/CD56- human
PBMCs are used as effector cells, CHO cells transfected with human
MCSP D3 as target cells. FIG. 16B) The macaque T cell line 4119
LnPx are used as effector cells, CHO cells transfected with
cynomolgus MCSP D3 as target cells. The assay is performed as
described in Example 11.
[0279] FIGS. 17A-17B
[0280] Cytotoxicity activity induced by designated cross-species
specific MCSP specific single chain constructs redirected to
indicated target cell lines. FIG. 17A) Stimulated CD4-/CD56- human
PBMCs are used as effector cells, CHO cells transfected with human
MCSP D3 as target cells. FIG. 17B) The macaque T cell line 4119
LnPx are used as effector cells, CHO cells transfected with
cynomolgus MCSP D3 as target cells. The assay is performed as
described in Example 11.
[0281] FIGS. 18(1)-18(3)
[0282] Plasma stability of MCSP and CD3 cross-species specific
bispecific single chain antibodies tested by the measurement of
cytotoxicity activity induced by samples of the designated single
chain constructs incubated with 50% human plasma at 37.degree. C.
and 4.degree. C. for 24 hours respectively or with addition of 50%
human plasma immediately prior to cytotoxicity testing or without
addition of plasma. CHO cells transfected with human MCSP are used
as target cell line and stimulated CD4-/CD56- human PBMCs are used
as effector cells. The assay is performed as described in Example
12.
[0283] FIGS. 19A-19F Initial drop and recovery (i.e.
redistribution) of absolute T cell counts (open squares), in
peripheral blood of B-NHL patients (patent numbers 1, 7, 23, 30,
31, and 33 of Table 4), who had essentially no circulating
CD19-positive target B cells (filled triangles), during the
starting phase of intravenous infusion with the CD3 binding
molecule CD19.times.CD3 recognizing a conventional context
dependent CD3 epitope. Absolute cell counts are given in 1000 cells
per microliter blood. The first data point shows baseline counts
immediately prior to the start of infusion. The CD19.times.CD3 dose
is given in parentheses beside the patient number.
[0284] FIGS. 20A-20B
[0285] (FIG. 20A) Repeated T cell redistribution (open squares) in
B-NHL patient #19 (Table 4) who had no circulating CD19-positive
target B cells (filled triangles) and developed CNS symptoms under
continuous intravenous infusion with CD19.times.CD3 at a starting
dose of 5 .mu.g/m.sup.2/24 h for one day followed by a sudden dose
increase to 15 .mu.g/m.sup.2/24 h. Absolute cell counts are given
in 1000 cells per microliter blood. The first data point shows
baseline counts immediately prior to the start of infusion. After
recovery of circulating T cells from the first episode of
redistribution triggered by the treatment start at 5
.mu.g/m.sup.2/24 h the stepwise dose increase from 5 to 15
.mu.g/m.sup.2/24 h triggered a second episode of T cell
redistribution that was associated with the development of CNS
symptoms dominated by confusion and disorientation.
[0286] (FIG. 20B) Repeated T cell redistribution in a B-NHL
patient, who developed CNS symptoms under repeated intravenous
bolus infusion with CD19.times.CD3 at 1.5 .mu.g/m.sup.2. Absolute
cell counts are given in 1000 cells per microliter blood. The
infusion time for each bolus administration was 2 to 4 hours.
Vertical arrows indicate the start of bolus infusions. Data points
at the beginning of each bolus administration show the T cell
counts immediately prior to start of bolus infusion. Each bolus
infusion triggered an episode of T cell redistribution followed by
recovery of the T cell counts prior to the next bolus infusion.
Finally the third episode of T cell redistribution was associated
with the development of CNS symptoms in this patient.
[0287] FIG. 21
[0288] Complex T cell redistribution pattern (open squares) in
B-NHL patient #20 (Table 4) without circulating CD19-positive
target B cells (filled triangles), during ramp initiation of the
CD19.times.CD3 infusion i.e. even gradual increase of flow-rate
from almost zero to 15 pg/m.sup.2/24 h during the first 24 hours of
treatment. Absolute cell counts are given in 1000 cells per
microliter blood. The first data point shows baseline counts
immediately prior to the start of infusion. The CD19.times.CD3 dose
is given in parentheses beside the patient number. T cells
reappearing in the circulating blood after the initial
redistribution triggered by the first exposure to CD19.times.CD3
are partially induced to redisappear from circulating blood again
by still increasing levels of CD19.times.CD3 during the ramp
phase.
[0289] FIG. 22
[0290] T and B cell counts during treatment with CD19.times.CD3 of
B-NHL patient #13 (Table 4) who had a significant number of
circulating CD19-positive target B (lymphoma) cells (filled
triangles). Absolute cell counts are given in 1000 cells per
microliter blood. The first data point shows baseline counts
immediately prior to the start of infusion. The CD19.times.CD3 dose
is given in parentheses beside the patient number. T cells (open
squares) disappear completely from the circulation upon start of
CD19.times.CD3 infusion and do not reappear until the circulating
CD19-positive B (lymphoma) cells (filled triangles) are depleted
from the peripheral blood.
[0291] FIG. 23
[0292] Repeated T cell redistribution (open squares) in B-NHL
patient #24 (Table 4), who had essentially no circulating
CD19-positive target B cells (filled triangles) and developed CNS
symptoms upon initiation of CD19.times.CD3 infusion without
additional HSA as required for stabilisation of the drug (upper
panel). After first recovery of circulating T cells from initial
redistribution the uneven drug flow due to the lack of stabilizing
HSA triggered a second episode of T cell redistribution that was
associated with the development of CNS symptoms dominated by
confusion and disorientation. When the same patient was restarted
correctly with CD19.times.CD3 solution containing additional HSA
for drug stabilisation, no repeated T cell redistribution was
observed (lower panel) and the patient did not again develop any
CNS symptoms. Absolute cell counts are given in 1000 cells per
microliter blood. The first data point shows baseline counts
immediately prior to the start of infusion. The CD19.times.CD3 dose
is given in parentheses beside the patient number.
[0293] FIG. 24
[0294] Model of T cell adhesion to endothelial cells induced by
monovalent binding to context dependent CD3 epitopes. Monovalent
interaction of a conventional CD3 binding molecule to its context
dependent epitope on CD3 epsilon can lead to an allosteric change
in the conformation of CD3 followed by the recruitment of Nck2 to
the cytoplasmic domain of CD3 epsilon (Gil et al. (2002) Cell 109:
901). As Nck2 is directly linked to integrins via PINCH and ILK
(Legate et al. (2006) Nat Rev Mol Cell Biol 7: 20), recruitment of
Nck2 to the cytoplasmic domain of CD3 epsilon following an
allosteric change in the conformation of CD3 through binding of a
conventional CD3 binding molecule (like the CD19.times.CD3 of
example 13) to its context dependent epitope on CD3 epsilon, can
increase the adhesiveness of T cells to endothelial cells by
transiently switching integrins on the T cell surface into their
more adhesive isoform via inside-out-signalling.
[0295] FIG. 25
[0296] Cytotoxic activity of CD33-AF5 VH-VL.times.I2C VH-VL test
material used for the in vivo study in cynomolgus monkeys as
described in Example 14. Specific lysis of CD33-positive target
cells was determined in a standard .sup.51Chromium release assay at
increasing concentrations of CD33-AF5 VH-VL.times.I2C VH-VL. Assay
duration was 18 hours. The macaque T cell line 4119 LnPx was used
as source of effector cells. CHO cells transfected with cynomolgus
CD33 served as target cells. Effector- to target cell ratio
(E:T-ratio) was 10:1. The concentration of CD33-AF5 VH-VL.times.I2C
VH-VL required for half-maximal target cell lysis (EC50) was
calculated from the dose response curve with a value of 2.7
ng/ml.
[0297] FIGS. 26A-26B(2)
[0298] (FIG. 26A) Dose- and time-dependent depletion of
CD33-positive monocytes from the peripheral blood of cynomolgus
monkeys through intravenous continuous infusion of CD33-AF5
VH-VL.times.I2C VH-VL as described in Example 14. The percentage
relative to baseline (i.e. 100%) of absolute circulating
CD33-positive monocyte counts after the duration of treatment as
indicated above the columns is shown for each of two cynomolgus
monkeys per dose level. The dose level (i.e. infusion flow-rate) is
indicated below the columns. No depletion of circulating
CD33-positive monocytes was observed in animals 1 and 2 treated for
7 days at a dose of 30 .mu.g/m.sup.2/24 h. In animals 3 and 4
treated for 7 days at a dose of 60 .mu.g/m.sup.2/24 h circulating
CD33-positive monocyte counts were reduced to 68% and 40% of
baseline, respectively. At 240 .mu.g/m.sup.2/24 h circulating
CD33-positive monocytes were almost completely depleted from the
peripheral blood after 3 days of treatment (animals 5 and 6). At
1000 .mu.g/m.sup.2/24 h depletion of circulating CD33-positive
monocytes from the peripheral blood was completed already after 1
day of treatment (animals 7 and 8).
[0299] (FIGS. 26B(1)-26B(2)) Course of T cell and CD33-monocyte
counts in peripheral blood of two cynomolgus monkeys during
continuous infusion of CD33-AF5 VH-VL.times.I2C VH-VL for 14 days
at 120 .mu.g/m.sup.2/24 h. Absolute cell counts are given in 1000
cells per microliter blood. The first data point shows baseline
counts immediately prior to the start of infusion. After initial
mobilisation of CD33-monocytes during the first 12 hours upon start
of infusion CD33-monocytes in peripheral blood (filled triangles)
are depleted by two thirds (animal 10) and 50% (animal 9) relative
to the respective baseline counts during the further course of
infusion. Circulating T cell counts (open squares) show a limited
initial drop followed by recovery still during the presence of
circulating CD33-positive monocytic target cells.
[0300] FIG. 27
[0301] Cytotoxic activity of MCSP-G4 VH-VL.times.I2C VH-VL test
material used for the in vivo study in cynomolgus monkeys as
described in Example 15. Specific lysis of MCSP-positive target
cells was determined in a standard .sup.51Chromium release assay at
increasing concentrations of MCSP-G4 VH-VL.times.I2C VH-VL. Assay
duration was 18 hours. The macaque T cell line 4119 LnPx was used
as source of effector cells. CHO cells transfected with cynomolgus
MCSP served as target cells. Effector- to target cell ratio
(E:T-ratio) was 10:1. The concentration of MCSP-G4 VH-VL.times.I2C
VH-VL required for half-maximal target cell lysis (EC50) was
calculated from the dose response curve with a value of 1.9
ng/ml.
[0302] FIGS. 28(1)(A)-28(2)(B)
[0303] Absence of initial episodes of drop and subsequent recovery
of absolute T cell counts (i.e. redistribution) in peripheral blood
of cynomolgus monkeys during the starting phase of intravenous
infusion with the CD3 binding molecule MCSP-G4 VH-VL.times.I2C
VH-VL recognizing an essentially context independent CD3 epitope.
Absolute cell counts are given in 1000 cells per microliter blood.
The first data point shows baseline counts immediately prior to the
start of infusion. The MCSP-G4 VH-VL.times.I2C VH-VL dose is given
in parentheses beside the animal number. In the known absence of
MCSP-positive target cells from the circulating blood of cynomolgus
monkeys there is no induction of T cell redistribution (i.e. an
initial episode of drop and subsequent recovery of absolute T cell
counts) through target cell mediated crosslinking of CD3. Moreover,
induction of T cell redistribution (i.e. an initial episode of drop
and subsequent recovery of absolute T cell counts) through a
signal, which the T cells may receive through exclusive interaction
with a CD3 binding site only, can be avoided by the use of CD3
binding molecules like MCSP-G4 VH-VL.times.I2C VH-VL recognizing an
essentially context independent CD3 epitope.
[0304] FIGS. 29(1)-29(12)
[0305] FACS binding analysis of designated cross-species specific
bispecific constructs to CHO cells transfected with human CD33, the
human CD3+ T cell line HPB-ALL, CHO cells transfected with macaque
CD33 and macaque PBMC respectively. The FACS staining is performed
as described in Example 16.4. The bold lines represent cells
incubated with 5 .mu.g/ml purified bispecific single chain
construct or cell culture supernatant of transfected cells
expressing the cross-species specific bispecific antibody
constructs. The filled histograms reflect the negative controls.
Supernatant of untransfected CHO cells was used as negative
control. For each cross-species specific bispecific single chain
construct the overlay of the histograms shows specific binding of
the construct to human and macaque CD33 and human and macaque
CD3.
[0306] FIGS. 30(1)-30(8)
[0307] The diagrams show results of chromium release assays
measuring cytotoxic activity induced by designated cross-species
specific CD33 specific single chain constructs redirected to the
indicated target cell lines. Effector cells were also used as
indicated. The assays are performed as described in Example 16.5.
The diagrams clearly demonstrate for each construct the potent
recruitment of cytotoxic activity of human and macaque effector
cells against human and macaque CD33 transfected CHO cells,
respectively.
[0308] FIG. 31
[0309] SDS PAGE gel and Western blot monitoring the purification of
the cross-species specific bispecific single chain molecule
designated E292F3 HL.times.I2C HL. Samples from the eluate, the
cell culture supernatant (SN) and the flow through of the column
(FT) were analyzed as indicated. A protein marker (M) was applied
as size reference. A strong protein band with a molecular weight
between 50 and 60 kDa in the SDS PAGE gel demonstrates the
efficient purification of the cross-species specific bispecific
single chain molecule to a very high degree of purity with the
one-step purification method described in Example 17.2. The Western
blot detecting the histidine.sub.6 tag confirms the identity of the
protein band in the eluate as the cross-species specific bispecific
single chain molecule. The faint signal for the flow through sample
in this sensitive detection method further shows the nearly
complete capture of bispecific single chain molecules by the
purification method.
[0310] FIG. 32
[0311] SDS PAGE gel and Western blot monitoring the purification of
the cross-species specific bispecific single chain molecule
designated V207C12 HL.times.H2C HL. Samples from the eluate, the
cell culture supernatant (SN) and the flow through of the column
(FT) were analyzed as indicated. A protein marker (M) was applied
as size reference. A strong protein band with a molecular weight
between 50 and 60 kDa in the SDS PAGE gel demonstrates the
efficient purification of the cross-species specific bispecific
single chain molecule to a very high degree of purity with the
one-step purification method described in Example 17.2. The Western
blot detecting the histidine.sub.6 tag confirms the identity of the
protein band in the eluate as the cross-species specific bispecific
single chain molecule. The faint signal for the flow through sample
in this sensitive detection method further shows the nearly
complete capture of bispecific single chain molecules by the
purification method.
[0312] FIG. 33
[0313] SDS PAGE gel and Western blot monitoring the purification of
the cross-species specific bispecific single chain molecule
designated AF5HL.times.F12QHL. Samples from the eluate, the cell
culture supernatant (SN) and the flow through of the column (FT)
were analyzed as indicated. A protein marker (M) was applied as
size reference. A strong protein band with a molecular weight
between 50 and 60 kDa in the SDS PAGE gel demonstrates the
efficient purification of the cross-species specific bispecific
single chain molecule to a very high degree of purity with the
one-step purification method described in Example 17.2. The Western
blot detecting the histidine.sub.6 tag confirms the identity of the
protein band in the eluate as the cross-species specific bispecific
single chain molecule. The signal in the flow through sample in
this sensitive detection method is explained by saturation of the
affinity column due to the high concentration of bispecific single
chain molecules in the supernatant.
[0314] FIG. 34A-34B
[0315] Standard curve of AF5HL.times.I2CHL in 50% macaque monkey
serum. The upper diagram shows the standard curve generated for the
assay as described in Example 18.2.
[0316] The lower diagram shows results for quality control samples
of AF5HL.times.I2CHL in 50% macaque monkey serum. The recovery
rates are above 90% for the high and mid QC sample and above 80%
for the low QC sample.
[0317] Thus the assay allows for detection of AF5HL.times.I2CHL in
serum samples in the range from 10 ng/ml to 200 ng/ml (before
dilution).
[0318] FIG. 35A-35B
[0319] Standard curve of MCSP-G4 HL.times.I2C HL in 50% macaque
monkey serum. The upper diagram shows the standard curve generated
for the assay as described in Example 18.2.
[0320] The lower diagram shows results for quality control samples
of MCSP-G4 HL.times.I2C HL in 50% macaque monkey serum. The
recovery rates are above 98% for the high and mid QC sample and
above 85% for the low QC sample.
[0321] Thus the assay allows for detection of MCSP-G4 HL.times.I2C
HL in serum samples in the range from 10 ng/ml to 200 ng/ml (before
dilution).
[0322] FIG. 36
[0323] FACS binding analysis of an anti-Flag antibody to CHO cells
transfected with the 1-27 N-terminal amino acids of CD3 epsilon of
the designated species fused to cynomolgus EpCAM. The FACS staining
was performed as described in Example 19.1. The bold lines
represent cells incubated with the anti-Flag antibody. The filled
histograms reflect the negative controls. PBS with 2% FCS was used
as negative control. The histograms show strong and comparable
binding of the anti-Flag antibody to all transfectants indicating
strong and equal expression of the transfected constructs.
[0324] FIG. 37
[0325] FACS binding analysis of the I2C IgG1 construct to CHO cells
expressing the 1-27 N-terminal amino acids of CD3 epsilon of the
designated species fused to cynomolgus EpCAM. The FACS staining is
performed as described in Example 19.3. The bold lines represent
cells incubated with 50 .mu.l cell culture supernatant of cells
expressing the I2C IgG1 construct. The filled histograms reflect
the negative control. Cells expressing the 1-27 N-terminal amino
acids of CD3 epsilon of swine fused to cynomolgus EpCAM were used
as negative control. In comparison with the negative control the
histograms clearly demonstrate binding of the I2C IgG1 construct to
1-27 N-terminal amino acids of CD3 epsilon of human, marmoset,
tamarin and squirrel monkey.
[0326] FIG. 38
[0327] FACS binding analysis of the I2C IgG1 construct as described
in Example 19.2 to human CD3 with and without N-terminal His6 tag
as described in Examples 6.1 and 5.1 respectively. The bold lines
represent cells incubated with the anti-human CD3 antibody UCHT-1,
the penta-His antibody (Qiagen) and cell culture supernatant of
cells expressing the I2C IgG1 construct respectively as indicated.
The filled histograms reflect cells incubated with an irrelevant
murine IgG1 antibody as negative control.
[0328] The upper two histogram overlays show comparable binding of
the UCHT-1 antibody to both transfectants as compared to the
isotype control demonstrating expression of both recombinant
constructs. The centre histogram overlays show binding of the penta
his antibody to the cells expressing the His6-human CD3 epsilon
chain (His6-CD3) but not to the cells expressing the wild-type CD3
epsilon chain (WT-CD3). The lower Histogram overlays show binding
of the I2C IgG1 construct to the wild-type human CD3 epsilon chain
but not to the His6-human CD3 epsilon chain. These results
demonstrate that a free N-terminus is essential for binding of the
cross-species specific anti-CD3 binding molecule I2C to the CD3
epsilon chain.
[0329] FIGS. 39A-39L
[0330] FACS binding analysis of designated cross-species specific
bispecific single chain constructs to CHO cells transfected with
human MCSP D3, the human CD3+ T cell line HPB-ALL, CHO cells
transfected with macaque MCSP D3 and the macaque T cell line 4119
LnPx respectively. The FACS staining was performed as described in
Example 10. The bold lines represents cells incubated with 2
.mu.g/ml purified bispecific single chain construct or cell
supernatant containing the bispecific single chain construct
respectively. The filled histograms reflect the negative controls.
Supernatant of untransfected CHO cells was used as negative control
for binding to the T cell lines. A single chain construct with
irrelevant target specificity was used as negative control for
binding to the MCSP D3 transfected CHO cells. For each
cross-species specific bispecific single chain construct the
overlay of the histograms shows specific binding of the construct
to human and macaque MCSP D3 and human and macaque CD3.
[0331] FIGS. 40A-40D
[0332] Cytotoxic activity induced by designated cross-species
specific MCSP D3 specific single chain constructs redirected to the
indicated target cell lines. Effector cells and effector to target
ratio were also used as indicated. The assay is performed as
described in Example 11. The diagrams clearly demonstrate potent
cross-species specific recruitment of cytotoxic activity by each
construct.
[0333] FIGS. 41A-41B
[0334] FACS binding analysis of designated cross-species specific
bispecific single chain constructs to CHO cells transfected with
human CD33, the human CD3+ T cell line HPB-ALL, CHO cells
transfected with macaque CD33 and macaque PBMC respectively. The
FACS staining was performed as described in Example 21.2. The bold
lines represent cells incubated with cell culture supernatant of
transfected cells expressing the cross-species specific bispecific
antibody constructs. The filled histograms reflect the negative
controls. Supernatant of untransfected CHO cells was used as
negative control. For each cross-species specific bispecific single
chain construct the overlay of the histograms shows specific
binding of the construct to human and macaque CD33 and human and
macaque CD3.
[0335] FIG. 42
[0336] The diagrams show results of chromium release assays
measuring cytotoxic activity induced by designated cross-species
specific CD33 specific single chain constructs redirected to the
indicated target cell lines. Effector cells were also used as
indicated. The assays are performed as described in Example 21.3.
The diagrams clearly demonstrate for each construct the potent
recruitment of cytotoxic activity of human and macaque effector
cells against human and macaque CD33 transfected CHO cells,
respectively.
[0337] FIG. 43
[0338] T cell redistribution in a chimpanzee under weekly
intravenous bolus infusion with PBS/5% HSA and PBS/5% HSA plus
single-chain EpCAM/CD3-bispecific antibody construct at doses of
1.6, 2.0, 3.0 and 4.5 .mu.g/kg. The infusion time for each bolus
administration was 2 hours. Vertical arrows indicate the start of
bolus infusions. Data points at the beginning of each bolus
administration show the T cell counts immediately prior to start of
bolus infusion. Each bolus infusion of the single-chain
EpCAM/CD3-bispecific antibody construct, which recognizes a
conventional context dependent CD3 epitope, triggered an episode of
T cell redistribution followed by recovery of T cells to baseline
values prior to the next bolus infusion.
[0339] FIG. 44
[0340] CD3 specific ELISA analysis of periplasmic preparations
containing Flag tagged scFv protein fragments from selected clones.
Periplasmic preparations of soluble scFv protein fragments were
added to wells of an ELISA plate, which had been coated with
soluble human CD3 epsilon (aa 1-27)-Fc fusion protein and had been
additionally blocked with PBS 3% BSA. Detection was performed by a
monoclonal anti Flag-Biotin-labeled antibody followed by
peroxidase-conjugated Streptavidin. The ELISA was developed by an
ABTS substrate solution. The OD values (y axis) were measured at
405 nm by an ELISA reader. Clone names are presented on the x
axis.
[0341] FIG. 45
[0342] ELISA analysis of periplasmic preparations containing Flag
tagged scFv protein fragments from selected clones. The same
periplasmic preparations of soluble scFv protein fragments as in
FIG. 44 were added to wells of an ELISA plate which had not been
coated with human CD3 epsilon (aa 1-27)-Fc fusion protein but with
hulgG1 (Sigma) and blocked with 3% BSA in PBS.
[0343] Detection was performed by a monoclonal anti
Flag-Biotin-labeled antibody followed by peroxidase-conjugated
Streptavidin. The ELISA was developed by an ABTS substrate
solution. The OD values (y axis) were measured at 405 nm by an
ELISA reader. Clone names are presented on the x axis.
[0344] FIGS. 46(1)-46(4)
[0345] FACS binding analysis of the designated cross-species
specific bispecific single chain constructs to CHO cells
transfected with the human PSMA, human CD3+ T cell line HPB-ALL,
CHO cells transfected with macaque PSMA and a macaque T cell line
4119 LnPx. The FACS staining is performed as described in Example
24.4. The thick line represents cells incubated with cell culture
supernatant that are subsequently incubated with the anti-his
antibody and the PE labeled detection antibody. The thin histogram
line reflects the negative control: cells only incubated with the
anti-his antibody and the detection antibody.
[0346] FIGS. 47(1)A-47(2)B
[0347] Cytotoxic activity induced by the designated cross-species
specific bispecific single chain constructs redirected to indicated
target cell lines. A) and B) Stimulated CD4-/CD56- human PBMCs are
used as effector cells, CHO cells transfected with human PSMA as
target cells. The assay is performed as described in Example
24.5.
[0348] FIGS. 48(1)A-48(2)B
[0349] Cytotoxic activity induced by the designated cross-species
specific bispecific single chain constructs redirected to indicated
target cell lines. A) and B) The macaque T cell line 4119 LnPx is
used as effector cells, CHO cells transfected with macaque PSMA as
target cells. The assay is performed as described in Example
24.5.
[0350] FIGS. 49A-49E
[0351] FACS binding analysis of the designated cross-species
specific bispecific single chain constructs to the human PSMA
positive prostate cancer cell line LNCaP, the human CD3+ T cell
line HPB-ALL and to the macaque T cell line 4119LnPx respectively.
The FACS staining was performed as described in Example 24.7. The
bold lines represent cells incubated with cell culture supernatant
of transfected cells expressing the cross-species specific
bispecific antibody constructs. The filled histograms reflect the
negative controls. Cell culture medium was used as a negative
control. For each cross-species specific bispecific single chain
construct shown the overlay of the histograms demonstrates binding
of the construct to human PSMA and human and macaque CD3.
[0352] FIGS. 50(1)-50(3)B
[0353] The diagrams show results of chromium release assays
measuring cytotoxic activity induced by designated cross-species
specific bispecific single chain constructs redirected to the
indicated target cell line. Effector cells were also used as
indicated. The assays were performed as described in Example 24.8.
The diagrams clearly demonstrate for the shown constructs the
potent recruitment of cytotoxic activity of human or macaque
effector T cells against PSMA-positive cancer cells by the example
of the human prostate cancer cell line LNCaP or the macaque cell
line 4119LnPx.
[0354] FIGS. 51A-51C
[0355] FACS binding analysis of the designated cross-species
specific bispecific single chain constructs to PSMA positive cells.
The FACS staining was performed as described in Example 24.7. For
each cross-species specific bispecific single chain construct shown
the overlay of the histograms demonstrates binding of the construct
to human PSMA and human and macaque CD3.
[0356] FIGS. 52A-52B
[0357] The diagrams show results of chromium release assays
measuring cytotoxic activity induced by designated cross-species
specific bispecific single chain constructs redirected to the
indicated target cell line. Effector cells were also used as
indicated. The assays were performed as described in Example 24.8.
The diagrams clearly demonstrate for the shown constructs the
potent recruitment of cytotoxic activity of human or macaque
effector T cells against PSMA-positive cells.
[0358] FIG. 53A-53D
[0359] FACS binding analysis of designated bispecific single chain
constructs to CHO cells expressing designated human/rat PSMA
chimeras as described in Example 25.1. The FACS staining was
performed as described in Example 25.2. The bold lines represent
cells incubated with cell culture supernatant of transfected cells
expressing the bispecific antibody constructs. The filled
histograms show the negative controls. Supernatant of untransfected
CHO cells was used as negative control. For each bispecific single
chain construct the overlays of the histograms show specific
binding of the construct to to the chimeric constructs
huPSMArat140-169, huPSMArat191-258, huPSMArat281-284,
huPSMArat683-690 and huPSMArat716-750. Compared with the signals
obtained for the other bispecific single chain construct there is a
clear lack of binding for the bispecific single chain antibody
constructs PM84-D7.times.I2C, PM29-G1.times.I2C and
PM49-B9.times.I2C to the chimeric construct huPSMArat300-344.
Furthermore compared with the signals obtained for the other
bispecific single chain constructs there is a clear lack of binding
for the bispecific single chain antibody construct
PM34-C7.times.I2C to the construct huPSMArat598-617.
[0360] FIG. 54
[0361] Binding of scFv MP9076-A9, the PSMA target binder of PSMA
BiTE antibody PM 76-A9.times.I2C to 15-mer peptides spanning over
the extracellular domain of human PSMA and overlapping with their
neighboring peptides by 14 amino acids. Peptide numbers are plotted
on the X-axis. ELISA signals using His detection are plotted on the
Y-axis.
[0362] FIG. 55
[0363] Binding of scFv MP9076-610, the PSMA target binder of PSMA
BiTE antibody PM 76-610.times.I2C to 15-mer peptides spanning over
the extracellular domain of human PSMA and overlapping with their
neighboring peptides by 14 amino acids. Peptide numbers are plotted
on the X-axis. ELISA signals using His detection are plotted on the
Y-axis.
[0364] FIG. 56
[0365] Binding of scFv F1-A10, the PSMA target binder of PSMA BiTE
antibody PM F1-A10.times.I2C to 15-mer peptides spanning over the
extracellular domain of human PSMA and overlapping with their
neighboring peptides by 14 amino acids. Peptide numbers are plotted
on the X-axis. ELISA signals using His detection are plotted on the
Y-axis.
[0366] FIG. 57
[0367] Potential dominant epitopes of scFvs MP 9076-A9, MP 9076-610
and F1-A10. The potential core binding amino acids in the
three-dimensional structure of human PSMA are encircled by a dotted
line. Color codes depict scFvs and the respective epitopes. The
crystal structure of human PSMA was reported by Davis et al. in
2005 (PNAS, 102: 5981-6).
[0368] The present invention is additionally described by way of
the following illustrative non-limiting examples that provide a
better understanding of the present invention and of its many
advantages.
EXAMPLES
[0369] 1. Identification of CD3Epsilon Sequences from Blood Samples
of Non-Human Primates
[0370] Blood samples of the following non-human primates were used
for CD3epsilon-identification: Callithrix jacchus, Saguinus oedipus
and Saimiris ciureus. Fresh heparin-treated whole blood samples
were prepared for isolating total cellular RNA according to
manufacturer's protocol (QIAamp RNA Blood Mini Kit, Qiagen). The
extracted mRNA was transcribed into cDNA according to published
protocols. In brief, 10 .mu.l of precipitated RNA was incubated
with 1.2 .mu.l of 10.times. hexanucleotide mix (Roche) at
70.degree. C. for 10 minutes and stored on ice. A reaction mix
consisting of 4 .mu.l of 5.times. superscript II buffer, 0.2 .mu.l
of 0.1M dithiothreitole, 0.8 .mu.l of superscript II (Invitrogen),
1.2 .mu.l of desoxyribonucleoside triphosphates (25 .mu.M), 0.8
.mu.l of RNase Inhibitor (Roche) and 1.8 .mu.l of DNase and RNase
free water (Roth) was added. The reaction mix was incubated at room
temperature for 10 minutes followed by incubation at 42.degree. C.
for 50 minutes and at 90.degree. C. for 5 minutes. The reaction was
cooled on ice before adding 0.8 .mu.l of RNaseH (1 U/.mu.l, Roche)
and incubated for 20 minutes at 37.degree. C.
[0371] The first-strand cDNAs from each species were subjected to
separate 35-cycle polymerase chain reactions using Taq DNA
polymerase (Sigma) and the following primer combination designed on
database research: forward primer 5'-AGAGTTCTGGGCCTCTGC-3' (SEQ ID
NO: 253); reverse primer 5'-CGGATGGGCTCATAGTCTG-3' (SEQ ID NO:
254);. The amplified 550 bp-bands were gel purified (Gel Extraction
Kit, Qiagen) and sequenced (Sequiserve, Vaterstetten/Germany, see
sequence listing).
TABLE-US-00001 CD3epsilon Callithrix jacchus Nucleotides (SEQ ID
NO: 1051) CAGGACGGTAATGAAGAAATGGGTGATACTACACAGAACCCATATAAAGT
TTCCATCTCAGGAACCACAGTAACACTGACATGCCCTCGGTATGATGGAC
ATGAAATAAAATGGCTCGTAAATAGTCAAAACAAAGAAGGTCATGAGGAC
CACCTGTTACTGGAGGACTTTTCGGAAATGGAGCAAAGTGGTTATTATGC
CTGCCTCTCCAAAGAGACTCCCGCAGAAGAGGCGAGCCATTATCTCTACC
TGAAGGCAAGAGTGTGTGAGAACTGCGTGGAGGTGGAT Amino acids (SEQ ID NO: 3)
QDGNEEMGDTTQNPYKVSISGTTVTLTCPRYDGHEIKWLVNSQNKEGHED
HLLLEDFSEMEQSGYYACLSKETPAEEASHYLYLKARVCENCVEVD CD3epsilon Saguinus
oedipus Nucleotides (SEQ ID NO: 1052)
CAGGACGGTAATGAAGAAATGGGTGATACTACACAGAACCCATATAAAGT
TTCCATCTCAGGAACCACAGTAACACTGACATGCCCTCGGTATGATGGAC
ATGAAATAAAATGGCTTGTAAATAGTCAAAACAAAGAAGGTCATGAGGAC
CACCTGTTACTGGAGGATTTTTCGGAAATGGAGCAAAGTGGTTATTATGC
CTGCCTCTCCAAAGAGACTCCCGCAGAAGAGGCGAGCCATTATCTCTACC
TGAAGGCAAGAGTGTGTGAGAACTGCGTGGAGGTGGAT Amino acids (SEQ ID NO: 5)
QDGNEEMGDTTQNPYKVSISGTTVTLTCPRYDGHEIKWLVNSQNKEGHED
HLLLEDFSEMEQSGYYACLSKETPAEEASHYLYLKARVCENCVEVD CD3epsilon Saimiris
ciureus Nucleotides (SEQ ID NO: 1053)
CAGGACGGTAATGAAGAGATTGGTGATACTACCCAGAACCCATATAAAGT
TTCCATCTCAGGAACCACAGTAACACTGACATGCCCTCGGTATGATGGAC
AGGAAATAAAATGGCTCGTAAATGATCAAAACAAAGAAGGTCATGAGGAC
CACCTGTTACTGGAAGATTTTTCAGAAATGGAACAAAGTGGTTATTATGC
CTGCCTCTCCAAAGAGACCCCCACAGAAGAGGCGAGCCATTATCTCTACC
TGAAGGCAAGAGTGTGTGAGAACTGCGTGGAGGTGGAT Amino acids (SEQ ID NO: 7)
QDGNEEIGDTTQNPYKVSISGTTVTLTCPRYDGQEIKWLVNDQNKEGHED
HLLLEDFSEMEQSGYYACLSKETPTEEASHYLYLKARVCENCVEVD
[0372] 2. Generation of Cross-Species Specific Single Chain
Antibody Fragments (scFv) Binding to the N-Terminal Amino Acids
1-27 of CD3Epsilon of Man and Different Non-Chimpanzee Primates
[0373] 2.1. Immunization of Mice Using the N-Terminus of CD3Epsilon
Separated from its Native CD3-Context by Fusion to a Heterologous
Soluble Protein
[0374] Ten weeks old F1 mice from balb/c.times.C57black crossings
were immunized with the CD3epsilon-Fc fusion protein carrying
themost N-terminal amino acids 1-27 of the mature CD3epsilon chain
(1-27 CD3-Fc) of man and/or saimiris ciureus. To this end 40 .mu.g
of the 1-27 CD3-Fc fusion protein with 10 nmol of a
thioate-modified CpG-Oligonucleotide (5'-tccatgacgttcctgatgct-3')
(SEQ ID No. 343) in 300 ul PBS were injected per mouse
intra-peritoneally. Mice receive booster immunizations after 21, 42
and optionally 63 days in the same way. Ten days after the first
booster immunization, blood samples were taken and antibody serum
titer against 1-27 CD3-Fc fusion protein iwa tested by ELISA.
Additionally, the titer against the CD3-positive human T cell line
HPBall was tested in flow cytometry according to standard
protocols. Serum titers were significantly higher in immunized than
in non-immunized animals.
[0375] 2.2. Generation of an Immune Murine Antibody scFv Library:
Construction of a Combinatorial Antibody Library and Phage
Display
[0376] Three days after the last injection the murine spleen cells
were harvested for the preparation of total RNA according to
standard protocols.
[0377] A library of murine immunoglobuline (Ig) light chain (kappa)
variable region (VK) and Ig heavy chain variable region (VH)
DNA-fragments was constructed by RT-PCR on murine spleen RNA using
VK-and VH specific primer. cDNA was synthesized according to
standard protocols.
[0378] The primers were designed in a way to give rise to a 5'-XhoI
and a 3'-BstEII recognition site for the amplified heavy chain
V-fragments and to a 5'-SacI and a 3'-SpeI recognition site for
amplified VK DNA fragments.
[0379] For the PCR-amplification of the VH DNA-fragments eight
different 5'-VH-family specific primers (MVH1(GC)AG GTG CAG CTC GAG
GAG TCA GGA CCT (SEQ ID No. 344); MVH2 GAG GTC CAG CTC GAG CAG TCT
GGA CCT (SEQ ID No. 345); MVH3 CAG GTC CAA CTC GAG CAG CCT GGG GCT
(SEQ ID No. 346); MVH4 GAG GTT CAG CTC GAG CAG TCT GGG GCA (SEQ ID
No. 347); MVH5 GA(AG) GTG AAG CTC GAG GAG TCT GGA GGA (SEQ ID No.
348); MVH6 GAG GTG AAG CTT CTC GAG TCT GGA GGT (SEQ ID No. 349);
MVH7 GAA GTG AAG CTC GAG GAG TCT GGG GGA (SEQ ID No. 350); MVH8 GAG
GTT CAG CTC GAG CAG TCT GGA GCT (SEQ ID No. 351)) were each
combined with one 3'-VH primer (3'MuVHBstEII tga gga gac ggt gac
cgt ggt ccc ttg gcc cca g (SEQ ID No. 352)); for the PCR
amplification of the VK-chain fragments seven different
5'-VK-family specific primers (MUVK1 CCA GTT CCG AGC TCG TTG TGA
CTC AGG AAT CT (SEQ ID No. 353); MUVK2 CCA GTT CCG AGC TCG TGT TGA
CGC AGC CGC CC (SEQ ID No. 354); MUVK3 CCA GTT CCG AGC TCG TGC TCA
CCC AGT CTC CA (SEQ ID No. 355); MUVK4 CCA GTT CCG AGC TCC AGA TGA
CCC AGT CTC CA (SEQ ID No. 356); MUVK5 CCA GAT GTG AGC TCG TGA TGA
CCC AGA CTC CA (SEQ ID No. 357); MUVK6 CCA GAT GTG AGC TCG TCA TGA
CCC AGT CTC CA (SEQ ID No. 358); MUVK7 CCA GTT CCG AGC TCG TGA TGA
CAC AGT CTC CA (SEQ ID No. 359)) were each combined with one 3'-VK
primer (3'MuVkHindIII/BsiW1 tgg tgc act agt cgt acg ttt gat ctc aag
ctt ggt ccc (SEQ ID No. 360)).
[0380] The following PCR program was used for amplification:
denaturation at 94.degree. C. for 20 sec; primer annealing at
52.degree. C. for 50 sec and primer extension at 72.degree. C. for
60 sec and 40 cycles, followed by a 10 min final extension at
72.degree. C.
[0381] 450 ng of the kappa light chain fragments (SacI-SpeI
digested) were ligated with 1400 ng of the phagemid pComb3H5Bhis
(SacI-SpeI digested; large fragment). The resulting combinatorial
antibody library was then transformed into 300 ul of
electrocompetent Escherichia coli XL1 Blue cells by electroporation
(2.5 kV, 0.2 cm gap cuvette, 25 uFD, 200 Ohm, Biorad gene-pulser)
resulting in a library size of more than 10.sup.7 independent
clones. After one hour of phenotype expression, positive
transformants were selected for carbenicilline resistance encoded
by the pComb3H5BHis vector in 100 ml of liquid super broth
(SB)-culture over night. Cells were then harvested by
centrifugation and plasmid preparation was carried out using a
commercially available plasmid preparation kit (Qiagen).
[0382] 2800 ng of this plasmid-DNA containing the VK-library
(XhoI-BstEII digested; large fragment) were ligated with 900 ng of
the heavy chain V-fragments (XhoI-BstEII digested) and again
transformed into two 300 ul aliquots of electrocompetent E. coli
XL1 Blue cells by electroporation (2.5 kV, 0.2 cm gap cuvette, 25
uFD, 200 Ohm) resulting in a total VH-VK scFv (single chain
variable fragment) library size of more than 10.sup.7 independent
clones.
[0383] After phenotype expression and slow adaptation to
carbenicillin, the E. coli cells containing the antibody library
were transferred into SB-Carbenicillin (50 ug/mL) selection medium.
The E. coli cells containing the antibody library wass then
infected with an infectious dose of 10.sup.12 particles of helper
phage VCSM13 resulting in the production and secretion of
filamentous M13 phage, wherein phage particle contains single
stranded pComb3H5BHis-DNA encoding a murine scFv-fragment and
displayed the corresponding scFv-protein as a translational fusion
to phage coat protein III. This pool of phages displaying the
antibody library was later used for the selection of antigen
binding entities.
[0384] 2.3. Phage Display Based Selection of CD3-Specific
Binders
[0385] The phage library carrying the cloned scFv-repertoire was
harvested from the respective culture supernatant by PEG8000/NaCl
precipitation and centrifugation. Approximately 10.sup.11 to
10.sup.12 scFv phage particles were resuspended in 0.4 ml of
PBS/0.1% BSA and incubated with 10.sup.5 to 10.sup.7 Jurkat cells
(a CD3-positive human T-cell line) for 1 hour on ice under slow
agitation. These Jurkat cells were grown beforehand in RPMI medium
enriched with fetal calf serum (10%), glutamine and
penicillin/streptomycin, harvested by centrifugation, washed in PBS
and resuspended in PBS/1% FCS (containing Na Azide). scFv phage
which do not specifically bind to the Jurkat cells were eliminated
by up to five washing steps with PBS/1% FCS (containing Na Azide).
After washing, binding entities were eluted from the cells by
resuspending the cells in HCl-glycine pH 2.2 (10 min incubation
with subsequent vortexing) and after neutralization with 2 M Tris
pH 12, the eluate was used for infection of a fresh uninfected E.
coli XL1 Blue culture (OD600>0.5). The E. coli culture
containing E. coli cells successfully transduced with a phagemid
copy, encoding a human scFv-fragment, were again selected for
carbenicillin resistance and subsequently infected with VCMS 13
helper phage to start the second round of antibody display and in
vitro selection. A total of 4 to 5 rounds of selections were
carried out, normally.
[0386] 2.4. Screening for CD3-Specific Binders
[0387] Plasmid DNA corresponding to 4 and 5 rounds of panning was
isolated from E. coli cultures after selection. For the production
of soluble scFv-protein, VH-VL-DNA fragments were excised from the
plasmids (XhoI-SpeI). These fragments were cloned via the same
restriction sites in the plasmid pComb3H5BFlag/His differing from
the original pComb3H5BHis in that the expression construct (e.g.
scFv) includes a Flag-tag (TGD YKDDDDK) between the scFv and the
His6-tag and the additional phage proteins were deleted. After
ligation, each pool (different rounds of panning) of plasmid DNA
was transformed into 100 .mu.l heat shock competent E. coli TG1 or
XLI blue and plated onto carbenicillin LB-agar. Single colonies
were picked into 100 ul of LB carb (50 ug/ml).
[0388] E. coli transformed with pComb3H5BHis containing a VL-and
VH-segment produce soluble scFv in sufficient amounts after
excision of the gene III fragment and induction with 1 mM IPTG. Due
to a suitable signal sequence, the scFv-chain was exported into the
periplasma where it folds into a functional conformation.
[0389] Single E. coli TG1 bacterial colonies from the
transformation plates were picked for periplasmic small scale
preparations and grown in SB-medium (e.g. 10 ml) supplemented with
20 mM MgCl2 and carbenicillin 50 .mu.g/ml (and re-dissolved in PBS
(e.g. 1 ml) after harvesting. By four rounds of freezing at
-70.degree. C. and thawing at 37.degree. C., the outer membrane of
the bacteria was destroyed by temperature shock and the soluble
periplasmic proteins including the scFvs were released into the
supernatant. After elimination of intact cells and cell-debris by
centrifugation, the supernatant containing the human anti-human
CD3-scFvs was collected and used for further examination.
[0390] 2.5. Identification of CD3-Specific Binders
[0391] Binding of the isolated scFvs was tested by flow cytometry
on eukaryotic cells, which on their surface express a heterologous
protein displaying at its N-terminus the first 27 N-terminal amino
acids of CD3epsilon.
[0392] As described in Example 4, the first amino acids 1-27 of the
N-terminal sequence of the mature CD3 epsilon chain of the human T
cell receptor complex (amino acid sequence:
QDGNEEMGGITQTPYKVSISGTTVILT SEQ ID NO: 2) were fused to the
N-terminus of the transmembrane protein EpCAM so that the
N-terminus was located at the outer cell surface. Additionally, a
FLAG epitope was inserted between the N-terminal 1-27 CD3epsilon
sequence and the EpCAM sequence. This fusion product was expressed
in human embryonic kidney (HEK) and chinese hamster ovary (CHO)
cells.
[0393] Eukaryotic cells displaying the 27 most N-terminal amino
acids of mature CD3epsilon of other primate species were prepared
in the same way for Saimiri ciureus (Squirrel monkey) (CD3epsilon
N-terminal amino acid sequence: QDGNEEIGDTTQNPYKVSISGTTVTLT SEQ ID
NO: 8), for Callithrix jacchus (CD3epsilon N-terminal amino acid
sequence: QDGNEEMGDTTQNPYKVSISGTTVTLT SEQ ID NO: 4) and for
Saguinus oedipus (CD3epsilon N-terminal amino acid sequence:
QDGNEEMGDTTQNPYKVSISGTTVTLT SEQ ID NO: 6).
[0394] For flow cytometry 2.5.times.10.sup.5 cells are incubated
with 50 ul supernatant or with 5 .mu.g/ml of the purified
constructs in 50 .mu.l PBS with 2% FCS. The binding of the
constructs was detected with an anti-His antibody (Penta-His
Antibody, BSA free, Qiagen GmbH, Hilden, FRG) at 2 .mu.g/ml in 50
.mu.l PBS with 2% FCS. As a second step reagent a
R-Phycoerythrin-conjugated affinity purified F(ab')2 fragment, goat
anti-mouse IgG (Fc-gamma fragment specific), diluted 1:100 in 50
.mu.l PBS with 2% FCS (Dianova, Hamburg, FRG) was used. The samples
were measured on a FACSscan (BD biosciences, Heidelberg, FRG).
[0395] Binding was always confirmed by flow cytometry as described
in the foregoing paragraph on primary T cells of man and different
primates (e.g. saimiris ciureus, callithrix jacchus, saguinus
oedipus).
[0396] 2.6. Generation of Human/Humanized Equivalents of Non-Human
CD3Epsilon Specific scFvs
[0397] The VH region of the murine anti-CD3 scFv was aligned
against human antibody germline amino acid sequences. The human
antibody germline VH sequence was chosen which has the closest
homology to the non-human VH and a direct alignment of the two
amino acid sequences was performed. There were a number of
framework residues of the non-human VH that differ from the human
VH framework regions ("different framework positions"). Some of
these residues may contribute to the binding and activity of the
antibody to its target.
[0398] To construct a library that contain the murine CDRs and at
every framework position that differs from the chosen human VH
sequence both possibilities (the human and the maternal murine
amino acid residue), degenerated oligonucleotides were synthesized.
These oligonucleotides incorporate at the differing positions the
human residue with a probability of 75% and the murine residue with
a probability of 25%. For one human VH e.g. six of these
oligonucleotides had to be synthesized that overlap in a terminal
stretch of approximately 20 nucleotides. To this end every second
primer was an antisense primer. Restriction sites needed for later
cloning within the oligonucleotides were deleted.
[0399] These primers may have a length of 60 to 90 nucleotides,
depending on the number of primers that were needed to span over
the whole V sequence.
[0400] These e.g. six primers were mixed in equal amounts (e.g. 1
.mu.l of each primer (primer stocks 20 to 100 .mu.M) to a 20 .mu.l
PCR reaction) and added to a PCR mix consisting of PCR buffer,
nucleotides and Taq polymerase. This mix was incubated at
94.degree. C. for 3 minutes, 65.degree. C. for 1 minute, 62.degree.
C. for 1 minute, 59.degree. C. for 1 minute, 56.degree. C. for 1
minute, 52.degree. C. for 1 minute, 50.degree. C. for 1 minute and
at 72.degree. C. for 10 minutes in a PCR cycler. Subsequently the
product was run in an agarose gel electrophoresis and the product
of a size from 200 to 400 isolated from the gel according to
standard methods.
[0401] This PCR product was then used as a template for a standard
PCR reaction using primers that incorporate N-terminal and
C-terminal suitable cloning restriction sites. The DNA fragment of
the correct size (for a VH approximately 350 nucleotides) was
isolated by agarose gel electrophoresis according to standard
methods. In this way sufficient VH DNA fragment was amplified. This
VH fragment was now a pool of VH fragments that have each one a
different amount of human and murine residues at the respective
differing framework positions (pool of humanized VH). The same
procedure was performed for the VL region of the murine anti-CD3
scFv (pool of humanized VL).
[0402] The pool of humanized VH was then combined with the pool of
humanized VL in the phage display vector pComb3H5Bhis to form a
library of functional scFvs from which--after display on
filamentous phage--anti-CD3 binders were selected, screened,
identified and confirmed as described above for the parental
non-human (murine) anti-CD3 scFv. Single clones were then analyzed
for favorable properties and amino acid sequence. Those scFvs which
were closest in amino acid sequence homology to human germline
V-segments are preferred particularly those wherein at least one
CDR among CDR I and II of VH and CDR I and II of VLkappa or CDR I
and II of VLlambda shows more than 80% amino acid sequence identity
to the closest respective CDR of all human germline V-segments.
Anti-CD3 scFvs were converted into recombinant bispecific single
chain antibodies as described in the following Examples 9, 16, and
24.
[0403] 3. Generation of a Recombinant Fusion Protein of the
N-Terminal Amino Acids 1-27 of the Human CD3 Epsilon Chain Fused to
the Fc-Part of an IgG1 (1-27 CD3-Fc).
[0404] 3.1. Cloning and Expression of 1-27 CD3-Fc
[0405] The coding sequence of the 1-27 N-terminal amino acids of
the human CD3 epsilon chain fused to the hinge and Fc gamma region
of human immunoglobulin IgG1 as well as an 6 Histidine Tag were
obtained by gene synthesis according to standard protocols (cDNA
sequence and amino acid sequence of the recombinant fusion protein
are listed under SEQ ID NOs 230 and 229). The gene synthesis
fragment was designed as to contain first a Kozak site for
eukaryotic expression of the construct, followed by an 19 amino
acid immunoglobulin leader peptide, followed in frame by the coding
sequence of the first 27 amino acids of the extracellular portion
of the mature human CD3 epsilon chain, followed in frame by the
coding sequence of the hinge region and Fc gamma portion of human
IgG1, followed in frame by the coding sequence of a 6 Histidine tag
and a stop codon (FIG. 1). The gene synthesis fragment was also
designed as to introduce restriction sites at the beginning and at
the end of the cDNA coding for the fusion protein. The introduced
restriction sites, EcoRI at the 5' end and SalI at the 3' end, are
utilized in the following cloning procedures. The gene synthesis
fragment was cloned via EcoRI and SalI into a plasmid designated
pEF-DHFR (pEF-DHFR is described in Mack et al. Proc. Natl. Acad.
Sci. USA 92 (1995) 7021-7025 and Raum et al. Cancer Immunol
Immunother 50 (2001) 141-150) following standard protocols. A
sequence verified plasmid was used for transfection in the
FreeStyle 293 Expression System (Invitrogen GmbH, Karlsruhe,
Germany) according to the manufacturers protocol. After 3 days cell
culture supernatants of the transfectants were harvested and tested
for the presence of the recombinant construct in an ELISA assay.
Goat anti-human IgG, Fc-gamma fragment specific antibody (obtained
from Jackson ImmunoResearch Europe Ltd., Newmarket, Suffolk, UK)
was diluted in PBS to 5 .mu.g/ml and coated with 100 .mu.l per well
onto a MaxiSorp 96-well ELISA plate (Nunc GmbH & Co. KG,
Wiesbaden, Germany) over night at 4.degree. C. Wells were washed
with PBS with 0.05% Tween 20 (PBS/Tween and blocked with 3% BSA in
PBS (bovine Albumin, fraction V, Sigma-Aldrich Chemie GmbH,
Taufkirchen, Germany) for 60 minutes at room temperature (RT).
Subsequently, wells were washed again PBS/Tween and then incubated
with cell culture supernatants for 60 minutes at RT. After washing
wells were incubated with a peroxidase conjugated anti-His6
antibody (Roche Diagnostics GmbH, Roche Applied Science, Mannheim,
Germany) diluted 1:500 in PBS with 1% BSA for 60 minutes at RT.
Subsequently, wells were washed with 200 .mu.l PBS/Tween and 100
.mu.l of the SIGMAFAST OPD (SIGMAFAST OPD [o-Phenylenediamine
dihydrochloride] substrate solution (Sigma-Aldrich Chemie GmbH,
Taufkirchen, Germany) was added according to the manufacturers
protocol. The reaction was stopped by adding 100 .mu.l M
H.sub.2SO.sub.4. Color reaction was measured on a PowerWaveX
microplate spectrophotometer (BioTek Instruments, Inc., Winooski,
Vt., USA) at 490 nm and subtraction of background absorption at 620
nm. As shown in FIG. 2 presence of the construct as compared to
irrelevant supernatant of mock-transfected HEK 293 cells used as
negative control was clearly detectable.
[0406] 3.2. Binding Assay of Cross-Species Specific Single Chain
Antibodies to 1-27 CD3-Fc.
[0407] Binding of crude preparations of periplasmatically expressed
cross-species specific single chain antibodies specific for CD3
epsilon to 1-27 CD3-Fc was tested in an ELISA assay. Goat
anti-human IgG, Fc-gamma fragment specific antibody (Jackson
ImmunoResearch Europe Ltd., Newmarket, Suffolk, UK) was diluted in
PBS to 5 .mu.g/ml and coated with 100 .mu.l per well onto a
MaxiSorp 96-well ELISA plate (Nunc GmbH & Co. KG, Wiesbaden,
Germany) over night at 4.degree. C. Wells were washed with PBS with
0.05% Tween 20 (PBS/Tween and blocked with PBS with 3% BSA (bovine
Albumin, fraction V, Sigma-Aldrich Chemie GmbH, Taufkirchen,
Germany) for 60 minutes at RT. Subsequently, wells were washed with
PBS/Tween and incubated with supernatants of cells expressing the
1-27 CD3-Fc construct for 60 minutes at RT. Wells were washed with
PBS/Tween and incubated with crude preparations of
periplasmatically expressed cross-species specific single-chain
antibodies as described above for 60 minutes at room temperature.
After washing with PBS/Tween wells were incubated with peroxidase
conjugated anti-Flag M2 antibody (Sigma-Aldrich Chemie GmbH,
Taufkirchen, Germany) diluted 1:10000 in PBS with 1% BSA for 60
minutes at RT. Wells were washed with PBS/Tween and incubated with
100 .mu.l of the SIGMAFAST OPD (OPD [o-Phenylenediamine
dihydrochloride] substrate solution (Sigma-Aldrich Chemie GmbH,
Taufkirchen, Germany) according to the manufacturers protocol.
Color reaction was stopped with 100 .mu.l 1 M H.sub.2SO.sub.4 and
measured on a PowerWaveX microplate spectrophotometer (BioTek
Instruments, Inc., Winooski, Vt., USA) at 490 nm and subtraction of
background absorption at 620 nm. Strong binding of cross-species
specific human single chain antibodies specific for CD3 epsilon to
the 1-27 CD3-Fc construct compared to a murine anti CD3
single-chain antibody was observed (FIG. 3).
[0408] 4. Generation of Recombinant Transmembrane Fusion Proteins
of the N-Terminal Amino Acids 1-27 of CD3 Epsilon from Different
Non-Chimpanzee Primates Fused to EpCAM from Cynomolgus Monkey (1-27
CD3-EpCAM).
[0409] 4.1. Cloning and Expression of 1-27 CD3-EpCAM CD3 epsilon
was isolated from different non-chimpanzee primates (marmoset,
tamarin, squirrel monkey) and swine. The coding sequences of the
1-27 N-terminal amino acids of CD3 epsilon chain of the mature
human, common marmoset (Callithrix jacchus), cottontop tamarin
(Saguinus oedipus), common squirrel monkey (Saimiri sciureus) and
domestic swine (Sus scrofa; used as negative control) fused to the
N-terminus of Flag tagged cynomolgus EpCAM were obtained by gene
synthesis according to standard protocols. cDNA sequence and amino
acid sequence of the recombinant fusion proteins are listed under
SEQ ID NOs 231 to 240). The gene synthesis fragments were designed
as to contain first a BsrGI site to allow fusion in correct reading
frame with the coding sequence of a 19 amino acid immunoglobulin
leader peptide already present in the target expression vector,
which is followed in frame by the coding sequence of the N-terminal
1-27 amino acids of the extracellular portion of the mature CD3
epsilon chains, which is followed in frame by the coding sequence
of a Flag tag and followed in frame by the coding sequence of the
mature cynomolgus EpCAM transmembrane protein (FIG. 4). The gene
synthesis fragments were also designed to introduce a restriction
site at the end of the cDNA coding for the fusion protein. The
introduced restriction sites BsrGI at the 5' end and SalI at the 3'
end, were utilized in the following cloning procedures. The gene
synthesis fragments were then cloned via BsrGI and SalI into a
derivative of the plasmid designated pEF DHFR (pEF-DHFR is
described in Mack et al. Proc. Natl. Acad. Sci. USA 92 (1995)
7021-7025), which already contained the coding sequence of the 19
amino acid immunoglobulin leader peptide following standard
protocols. Sequence verified plasmids were used to transiently
transfect 293-HEK cells using the MATra-A Reagent (IBA GmbH,
Gottingen, Germany) and 12 .mu.g of plasmid DNA for adherent
293-HEK cells in 175 ml cell culture flasks according to the
manufacturers protocol. After 3 days of cell culture the
transfectants were tested for cell surface expression of the
recombinant transmembrane protein via an FACS assay according to
standard protocols. For that purpose a number of 2.5.times.10.sup.5
cells were incubated with the anti-Flag M2 antibody (Sigma-Aldrich
Chemie GmbH, Taufkirchen, Germany) at 5 .mu.g/ml in PBS with 2%
FCS. Bound antibody was detected with an R-Phycoerythrin-conjugated
affinity purified F(ab')2 fragment, goat anti-mouse IgG, Fc-gamma
fragment specific 1:100 in PBS with 2% FCS (Jackson ImmunoResearch
Europe Ltd., Newmarket, Suffolk, UK). The samples were measured on
a FACScalibur (BD biosciences, Heidelberg, Germany). Expression of
the Flag tagged recombinant transmembrane fusion proteins
consisting of cynomolgus EpCAM and the 1-27 N-terminal amino acids
of the human, marmoset, tamarin, squirrel monkey and swine CD3
epsilon chain respectively on transfected cells was clearly
detectable (FIG. 5).
[0410] 4.2. Binding of Cross-Species Specific Anti-CD3 Single Chain
Antibodies to the 1-27 CD3-EpCAM
[0411] Binding of crude preparations of periplasmatically expressed
cross-species specific anti CD3 single-chain antibodies to the 1-27
N-terminal amino acids of the human, marmoset, tamarin and squirrel
monkey CD3 epsilon chains respectively fused to cynomolgus Ep-CAM
was tested in an FACS assay according to standard protocols. For
that purpose a number of 2.5.times.10.sup.5 cells were incubated
with crude preparations of periplasmatically expressed
cross-species specific anti CD3 single-chain antibodies
(preparation was performed as described above and according to
standard protocols) and a single-chain murine anti-human CD3
antibody as negative control. As secondary antibody the Penta-His
antibody (Qiagen GmbH, Hildesheim, Germany) was used at 5 .mu.g/ml
in 50 .mu.l PBS with 2% FCS. The binding of the antibody was
detected with an R-Phycoerythrin-conjugated affinity purified
F(ab')2 fragment, goat anti-mouse IgG, Fc-gamma fragment specific,
diluted 1:100 in PBS with 2% FCS (Jackson ImmunoResearch Europe
Ltd., Newmarket, Suffolk, UK). The samples were measured on a
FACScalibur (BD biosciences, Heidelberg, Germany). As shown in
FIGS. 6A to 6E binding of single chain antibodies to the
transfectants expressing the recombinant transmembrane fusion
proteins consisting of the 1-27 N-terminal amino acids of CD3
epsilon of the human, marmoset, tamarin or squirrel monkey fused to
cynomolgus EpCAM was observed. No binding of cross-species specific
single chain antibodies was observed to a fusion protein consisting
of the 1-27 N-terminal CD3 epsilon of swine fused to cynomolgus
EpCAM used as negative control. Multi-primate cross-species
specificity of the anti-CD3 single chain antibodies was shown.
Signals obtained with the anti Flag M2 antibody and the
cross-species specific single chain antibodies were comparable,
indicating a strong binding activity of the cross-species specific
single chain antibodies to the N-terminal amino acids 1-27 of CD3
epsilon.
[0412] 5. Binding Analysis of Cross-Species Specific Anti-CD3
Single Chain Antibodies by Alanine-Scanning of Mouse Cells
Transfected with the Human CD3 Epsilon Chain and its Alanine
Mutants
[0413] 5.1. Cloning and Expression of Human Wild-Type CD3
Epsilon
[0414] The coding sequence of the human CD3 epsilon chain was
obtained by gene synthesis according to standard protocols (cDNA
sequence and amino acid sequence of the human CD3 epsilon chain are
listed under SEQ ID NOs 242 and 241). The gene synthesis fragment
was designed as to contain a Kozak site for eukaryotic expression
of the construct and restriction sites at the beginning and the end
of the cDNA coding for human CD3 epsilon. The introduced
restriction sites EcoRI at the 5' end and SalI at the 3' end, were
utilized in the following cloning procedures. The gene synthesis
fragment was then cloned via EcoRI and SalI into a plasmid
designated pEF NEO following standard protocols. pEF NEO was
derived of pEF DHFR (Mack et al. Proc. Natl. Acad. Sci. USA 92
(1995) 7021-7025) by replacing the cDNA of the DHFR with the cDNA
of the neomycin resistance by conventional molecular cloning. A
sequence verified plasmid was used to transfect the murine T cell
line EL4 (ATCC No. TIB-39) cultivated in RPMI with stabilized
L-glutamine supplemented with 10% FCS, 1% penicillin/streptomycin,
1% HEPES, 1% pyruvate, 1% non-essential amino acids (all Biochrom
AG Berlin, Germany) at 37.degree. C., 95% humidity and 7% CO.sub.2.
Transfection was performed with the SuperFect Transfection Reagent
(Qiagen GmbH, Hilden, Germany) and 2 .mu.g of plasmid DNA according
to the manufacturer's protocol. After 24 hours the cells were
washed with PBS and cultivated again in the aforementioned cell
culture medium with600 .mu.g/ml G418 for selection (PAA
Laboratories GmbH, Pasching, Austria). 16 to 20 days after
transfection the outgrowth of resistant cells was observed. After
additional 7 to 14 days cells were tested for expression of human
CD3 epsilon by FACS analysis according to standard protocols.
2.5.times.10.sup.5 cells were incubated with anti-human CD3
antibody UCHT-1 (BD biosciences, Heidelberg, Germany) at 5 .mu.g/ml
in PBS with 2% FCS. The binding of the antibody was detected with
an R-Phycoerythrin-conjugated affinity purified F(ab')2 fragment,
goat anti-mouse IgG, Fc-gamma fragment specific, diluted 1:100 in
PBS with 2% FCS (Jackson ImmunoResearch Europe Ltd., Newmarket,
Suffolk, UK). The samples were measured on a FACSCalibur (BD
biosciences, Heidelberg, Germany). Expression of human wild-type
CD3 on transfected EL4 cells is shown in FIG. 7.
[0415] 5.2. Cloning and Expression of the Cross-Species Specific
Anti-CD3 Single Chain Antibodies as IgG1 Antibodies
[0416] In order to provide improved means of detection of binding
of the cross-species specific single chain anti-CD3 antibodies H2C
HLP, A2J HLP and E2M HLP were converted into IgG1 antibodies with
murine IgG1 and human lambda constant regions. cDNA sequences
coding for the heavy and light chains of respective IgG antibodies
were obtained by gene synthesis according to standard protocols.
The gene synthesis fragments for each specificity were designed as
to contain first a Kozak site to allow eukaryotic expression of the
construct, which is followed by an 19 amino acid immunoglobulin
leader peptide (SEQ ID NOs 244 and 243), which is followed in frame
by the coding sequence of the respective heavy chain variable
region or respective light chain variable region, followed in frame
by the coding sequence of the heavy chain constant region of murine
IgG1 (SEQ ID NOs 246 and 245) or the coding sequence of the human
lambda light chain constant region (SEQ ID NO 248 and 247),
respectively. Restriction sites were introduced at the beginning
and the end of the cDNA coding for the fusion protein. Restriction
sites EcoRI at the 5' end and SalI at the 3' end were used for the
following cloning procedures. The gene synthesis fragments were
cloned via EcoRI and SalI into a plasmid designated pEF DHFR (Mack
et al. Proc. Natl. Acad. Sci. USA 92 (1995) 7021-7025) for the
heavy chain constructs and pEF ADA (pEF ADA is described in Raum et
al., Cancer Immunol Immunother., 50(3), (2001), 141-50) for the
light chain constructs) according to standard protocols. Sequence
verified plasmids were used for co-transfection of respective light
and heavy chain constructs in the FreeStyle 293 Expression System
(Invitrogen GmbH, Karlsruhe, Germany) according to the
manufacturers protocol. After 3 days cell culture supernatants of
the transfectants were harvested and used for the alanine-scanning
experiment.
[0417] 5.3. Cloning and Expression of Alanine Mutants of Human CD3
Epsilon for Alanine-Scanning
[0418] 27 cDNA fragments coding for the human CD3 epsilon chain
with an exchange of one codon of the wild-type sequence of human
CD3 epsilon into a codon coding for alanine (GCC) for each amino
acid of amino acids 1-27 of the extracellular domain of the mature
human CD3 epsilon chain respectively were obtained by gene
synthesis. Except for the exchanged codon the cDNA fragments were
identical to the aforementioned human wild-type CD3 cDNA fragment.
Only one codon was replaced in each construct compared to the human
wild-type CD3 cDNA fragment described above. Restriction sites
EcoRI and SalI were introduced into the cDNA fragments at identical
positions compared to the wild-type construct. All alanine-scanning
constructs were cloned into pEF NEO and sequence verified plasmids
were transfected into EL4 cells. Transfection and selection of
transfectants was performed as described above. As result a panel
of expressed constructs was obtained wherein the first amino acid
of the human CD3 epsilon chain, glutamine (Q, Gln) at position 1
was replaced by alanine. The last amino acid replaced by alanine
was the threonine (T, Thr) at position 27 of mature human wild-type
CD3 epsilon. For each amino acid between glutamine 1 and threonine
27 respective transfectants with an exchange of the wild-type amino
acid into alanine were generated.
[0419] 5.4. Alanine-Scanning Experiment
[0420] Chimeric IgG antibodies as described in 5.2 and
cross-species specific single chain antibodies specific for CD3
epsilon were tested in alanine-scanning experiment. Binding of the
antibodies to the EL4 cell lines transfected with the
alanine-mutant constructs of human CD3 epsilon as described in 5.3
was tested by FACS assay according to standard protocols.
2.5.times.10.sup.5 cells of the respective transfectants were
incubated with 50 .mu.l of cell culture supernatant containing the
chimeric IgG antibodies or with 50 .mu.l of crude preparations of
periplasmatically expressed single-chain antibodies. For samples
incubated with crude preparations of periplasmatically expressed
single-chain antibodies the anti-Flag M2 antibody (Sigma-Aldrich
Chemie GmbH, Taufkirchen, Germany) was used as secondary antibody
at 5 .mu.g/ml in 50 .mu.l PBS with 2% FCS. For samples incubated
with the chimeric IgG antibodies a secondary antibody was not
necessary. For all samples the binding of the antibody molecules
was detected with an R-Phycoerythrin-conjugated affinity purified
F(ab')2 fragment, goat anti-mouse IgG, Fc-gamma fragment specific,
diluted 1:100 in PBS with 2% FCS (Jackson ImmunoResearch Europe
Ltd., Newmarket, Suffolk, UK). Samples were measured on a
FACSCalibur (BD biosciences, Heidelberg, Germany). Differential
binding of chimeric IgG molecules or cross-species specific
single-chain antibodies to the EL4 cell lines transfected with the
alanine-mutants of human CD3 epsilon was detected. As negative
control either an isotype control or a crude preparation of a
periplasmatically expressed single-chain antibody of irrelevant
specificity was used respectively. UCHT-1 antibody was used as
positive control for the expression level of the alanine-mutants of
human CD3 epsilon. The EL4 cell lines transfected with the
alanine-mutants for the amino acids tyrosine at position 15, valine
at position 17, isoleucine at position 19, valine at position 24 or
leucine at position 26 of the mature CD3 epsilon chain were not
evaluated due to very low expression levels (data not shown).
Binding of the cross-species specific single chain antibodies and
the single chain antibodies in chimeric IgG format to the EL4 cell
lines transfected with the alanine-mutants of human CD3 epsilon is
shown in FIGS. 8A-8D as relative binding in arbitrary units with
the geometric mean fluorescence values of the respective negative
controls subtracted from all respective geometric mean fluorescence
sample values. To compensate for different expression levels all
sample values for a certain transfectant were then divided through
the geometric mean fluorescence value of the UCHT-1 antibody for
the respective transfectant. For comparison with the wild-type
sample value of a specificity all sample values of the respective
specificity were finally divided through the wild-type sample
value, thereby setting the wild-type sample value to 1 arbitrary
unit of binding.
[0421] The calculations used are shown in detail in the following
formula:
value_Sample ( x , y ) = Sample ( x , y ) - neg_Contr . ( x ) (
UCHT - 1 ( x ) - neg_Contr . ( x ) ) * WT ( y ) - neg_Contr . ( wt
) UCHT - 1 ( wt ) - neg_Contr . ( wt ) ##EQU00002##
[0422] In this equation value_Sample means the value in arbitrary
units of binding depicting the degree of binding of a specific
anti-CD3 antibody to a specific alanine-mutant as shown in FIGS.
8A-8D, Sample means the geometric mean fluorescence value obtained
for a specific anti-CD3 antibody assayed on a specific
alanine-scanning transfectant, neg_Contr. means the geometric mean
fluorescence value obtained for the negative control assayed on a
specific alanine-mutant, UCHT-1 means the geometric mean
fluorescence value obtained for the UCHT-1 antibody assayed on a
specific alanine-mutant, WT means the geometric mean fluorescence
value obtained for a specific anti-CD3 antibody assayed on the
wild-type transfectant, x specifies the respective transfectant, y
specifies the respective anti-CD3 antibody and wt specifies that
the respective transfectant is the wild-type.
[0423] As can be seen in FIGS. 8A-8D the IgG antibody A2J HLP
showed a pronounced loss of binding for the amino acids asparagine
at position 4, threonine at position 23 and isoleucine at position
25 of the mature CD3 epsilon chain. A complete loss of binding of
IgG antibody A2J HLP was observed for the amino acids glutamine at
position 1, aspartate at position 2, glycine at position 3 and
glutamate at position 5 of the mature CD3 epsilon chain. IgG
antibody E2M HLP showed a pronounced loss of binding for the amino
acids asparagine at position 4, threonine at position 23 and
isoleucine at position 25 of the mature CD3 epsilon chain. IgG
antibody E2M HLP showed a complete loss of binding for the amino
acids glutamine at position 1, aspartate at position 2, glycine at
position 3 and glutamate at position 5 of the mature CD3 epsilon
chain. IgG antibody H2C HLP showed an intermediate loss of binding
for the amino acid asparagine at position 4 of the mature CD3
epsilon chain and it showed a complete loss of binding for the
amino acids glutamine at position 1, aspartate at position 2,
glycine at position 3 and glutamate at position 5 of the mature CD3
epsilon chain. Single chain antibody F12Q HLP showed an essentially
complete loss of binding for the amino acids glutamine at position
1, aspartate at position 2, glycine at position 3 of the mature CD3
epsilon chain and glutamate at position 5 of the mature CD3 epsilon
chain.
[0424] 6. Binding Analysis of the Cross-Species Specific Anti-CD3
Binding Molecule H2C HLP to the Human CD3 Epsilon Chain with and
without N-Terminal His6 Tag Transfected into the Murine T Cell Line
EL4
[0425] 6.1. Cloning and Expression of the Human CD3 Epsilon Chain
with N-Terminal Six Histidine Tag (His6 Tag)
[0426] A cDNA fragment coding for the human CD3 epsilon chain with
a N-terminal His6 tag was obtained by gene synthesis. The gene
synthesis fragment was designed as to contain first a Kozak site
for eukaryotic expression of the construct, which is followed in
frame by the coding sequence of a 19 amino acid immunoglobulin
leader peptide, which is followed in frame by the coding sequence
of a His6 tag which is followed in frame by the coding sequence of
the mature human CD3 epsilon chain (the cDNA and amino acid
sequences of the construct are listed as SEQ ID NOs 256 and 255).
The gene synthesis fragment was also designed as to contain
restriction sites at the beginning and the end of the cDNA. The
introduced restriction sites EcoRI at the 5' end and SalI at the 3'
end, were used in the following cloning procedures. The gene
synthesis fragment was then cloned via EcoRI and SalI into a
plasmid designated pEF-NEO (as described above) following standard
protocols. A sequence verified plasmid was used to transfect the
murine T cell line EL4. Transfection and selection of the
transfectants were performed as described above. After 34 days of
cell culture the transfectants were used for the assay described
below. 6.2. Binding of the Cross-Species Specific Anti-CD3 Binding
Molecule H2C HLP to the Human CD3 Epsilon Chain with and without
N-Terminal His6 Tag
[0427] A chimeric IgG antibody with the binding specificity H2C HLP
specific for CD3 epsilon was tested for binding to human CD3
epsilon with and without N-terminal His6 tag. Binding of the
antibody to the EL4 cell lines transfected the His6-human CD3
epsilon and wild-type human CD3 epsilon respectively was tested by
an FACS assay according to standard protocols. 2.5.times.10.sup.5
cells of the transfectants were incubated with 50 .mu.l of cell
culture supernatant containing the chimeric IgG antibody or 50
.mu.l of the respective control antibodies at 5 .mu.g/ml in PBS
with 2% FCS. As negative control an appropriate isotype control and
as positive control for expression of the constructs the CD3
specific antibody UCHT-1 were used respectively. The binding of the
antibodies was detected with a R-Phycoerythrin-conjugated affinity
purified F(ab')2 fragment, goat anti-mouse IgG, Fc-gamma fragment
specific, diluted 1:100 in PBS with 2% FCS (Jackson ImmunoResearch
Europe Ltd., Newmarket, Suffolk, UK). Samples were measured on a
FACSCalibur (BD biosciences, Heidelberg, Germany). Compared to the
EL4 cell line transfected with wild-type human CD3 epsilon a clear
loss of binding of the chimeric IgG with binding specificity H2C
HLP to human-CD3 epsilon with an N-terminal His6 tag was detected.
These results showed that a free N-terminus of CD3 epsilon is
essential for binding of the cross-species specific anti-CD3
binding specificity H2C HLP to the human CD3 epsilon chain (FIG.
9).
[0428] 7. Cloning and Expression of the C-Terminal, Transmembrane
and Truncated Extracellular Domains of Human MCSP
[0429] The coding sequence of the C-terminal, transmembrane and
truncated extracellular domain of human MCSP (amino acids
1538-2322) was obtained by gene synthesis according to standard
protocols (cDNA sequence and amino acid sequence of the recombinant
construct for expression of the C-terminal, transmembrane and
truncated extracellular domain of human MCSP (designated as human
D3) are listed under SEQ ID NOs 250 and 249). The gene synthesis
fragment was designed as to contain first a Kozak site to allow
eukaryotic expression of the construct followed by the coding
sequence of an 19 amino acid immunoglobulin leader peptide followed
in frame by a FLAG tag, followed in frame by a sequence containing
several restriction sites for cloning purposes and coding for a 9
amino acid artificial linker (SRTRSGSQL), followed in frame by the
coding sequence of the C-terminal, transmembrane and truncated
extracellular domain of human MCSP and a stop codon. Restriction
sites were introduced at the beginning and at the end of the DNA
fragment. The restriction sites EcoRI at the 5' end and SalI at the
3' end were used in the following cloning procedures. The fragment
was digested with EcoRI and SalI and cloned into pEF-DHFR (pEF-DHFR
is described in Mack et al. Proc. Natl. Acad. Sci. USA 92 (1995)
7021-7025) following standard protocols. A sequence verified
plasmid was used to transfect CHO/dhfr- cells (ATCC No. CRL 9096).
Cells were cultivated in RPMI 1640 with stabilized glutamine,
supplemented with 10% FCS, 1% penicillin/streptomycin (all obtained
from Biochrom AG Berlin, Germany) and nucleosides from a stock
solution of cell culture grade reagents (Sigma-Aldrich Chemie GmbH,
Taufkirchen, Germany) to a final concentration of 10 .mu.g/ml
Adenosine, 10 .mu.g/ml Deoxyadenosine and 10 .mu.g/ml Thymidine, in
an incubator at 37.degree. C., 95% humidity and 7% CO.sub.2.
Transfection was performed using the PolyFect Transfection Reagent
(Qiagen GmbH, Hilden, Germany) and 5 .mu.g of plasmid DNA according
to the manufacturer's protocol. After cultivation for 24 hours
cells were washed once with PBS and cultivated again in RPMI 1640
with stabilized glutamine and 1% penicillin/streptomycin. Thus the
cell culture medium did not contain nucleosides and thereby
selection was applied on the transfected cells. Approximately 14
days after transfection the outgrowth of resistant cells was
observed. After an additional 7 to 14 days the transfectants were
tested for expression of the construct by FACS analysis.
2.5.times.10.sup.5 cells were incubated with 50 .mu.l of an
anti-Flag-M2 antibody (Sigma-Aldrich Chemie GmbH, Taufkirchen,
Germany) diluted to 5 .mu.g/ml in PBS with 2% FCS. The binding of
the antibody was detected with a R-Phycoerythrin-conjugated
affinity purified F(ab')2 fragment, goat anti-mouse IgG, Fc-gamma
fragment specific diluted 1:100 in PBS with 2% FCS (ImmunoResearch
Europe Ltd., Newmarket, Suffolk, UK). The samples were measured on
a FACScalibur (BD biosciences, Heidelberg, Germany).
[0430] 8. Cloning and Expression of the C-Terminal, Transmembrane
and Truncated Extracellular Domains of Macaque MCSP
[0431] The cDNA sequence of the C-terminal, transmembrane and
truncated extracellular domains of macaque MCSP (designated as
macaque D3) was obtained by a set of three PCRs on macaque skin
cDNA (Cat No. C1534218-Cy-BC; BioCat GmbH, Heidelberg, Germany)
using the following reaction conditions: 1 cycle at 94.degree. C.,
3 min., 40 cycles with 94.degree. C. for 0.5 min., 52.degree. C.
for 0.5 min. and 72.degree. C. for 1.75 min., terminal cycle of
72.degree. C. for 3 min. The following primers were used:
TABLE-US-00002 forward primer: (SEQ ID No. 361)
5'-GATCTGGTCTACACCATCGAGC-3' reverse primer: (SEQ ID No. 362)
5'-GGAGCTGCTGCTGGCTCAGTGAGG-3' forward primer: (SEQ ID No. 363)
5'-TTCCAGCTGAGCATGTCTGATGG-3' reverse primer: (SEQ ID No. 364)
5'-CGATCAGCATCTGGGCCCAGG-3' forward primer: (SEQ ID No. 365)
5'-GTGGAGCAGTTCACTCAGCAGGACC-3' reverse primer: (SEQ ID No. 366)
5'-GCCTTCACACCCAGTACTGGCC-3'
[0432] Those PCRs generated three overlapping fragments (A: 1-1329,
B: 1229-2428, C: 1782-2547) which were isolated and sequenced
according to standard protocols using the PCR primers and thereby
provided a 2547 bp portion of the cDNA sequence of macaque MCSP
(the cDNA sequence and amino acid sequence of this portion of
macaque MCSP are listed under SEQ ID NOs 252 and 251) from 74 bp
upstream of the coding sequence of the C-terminal domain to 121 bp
downstream of the stop codon. Another PCR using the following
reaction conditions: 1 cycle at 94.degree. C. for 3 min, 10 cycles
with 94.degree. C. for 1 min, 52.degree. C. for 1 min and
72.degree. C. for 2.5 min, terminal cycle of 72.degree. C. for 3
min was used to fuse the PCR products of the aforementioned
reactions A and B. The following primers are used:
TABLE-US-00003 forward primer: (SEQ ID No. 367)
5'-tcccgtacgagatctggatcccaattggatggcggactcgtgctgtt ctcacacagagg-3'
reverse primer: (SEQ ID No. 368)
5'-agtgggtcgactcacacccagtactggccattcttaagggcaggg-3'
[0433] The primers for this PCR were designed to introduce
restriction sites at the beginning and at the end of the cDNA
fragment coding for the C-terminal, transmembrane and truncated
extracellular domains of macaque MCSP. The introduced restriction
sites MfeI at the 5' end and SalI at the 3' end, were used in the
following cloning procedures. The PCR fragment was then cloned via
MfeI and SalI into a Bluescript plasmid containing the EcoRI/MfeI
fragment of the aforementioned plasmid pEF-DHFR (pEF-DHFR is
described in Raum et al. Cancer Immunol Immunother 50 (2001)
141-150) by replacing the C-terminal, transmembrane and truncated
extracellular domains of human MCSP. The gene synthesis fragment
contained the coding sequences of the immunoglobulin leader peptide
and the Flag tag as well as the artificial linker (SRTRSGSQL) in
frame to the 5' end of the cDNA fragment coding for the C-terminal,
transmembrane and truncated extracellular domains of macaque MCSP.
This vector was used to transfect CHO/dhfr- cells (ATCC No. CRL
9096). Cells were cultivated in RPMI 1640 with stabilized glutamine
supplemented with 10% FCS, 1% penicillin/streptomycin (all from
Biochrom AG Berlin, Germany) and nucleosides from a stock solution
of cell culture grade reagents (Sigma-Aldrich Chemie GmbH,
Taufkirchen, Germany) to a final concentration of 10 .mu.g/ml
Adenosine, 10 .mu.g/ml Deoxyadenosine and 10 .mu.g/ml Thymidine, in
an incubator at 37.degree. C., 95% humidity and 7% CO2.
Transfection was performed with PolyFect Transfection Reagent
(Qiagen GmbH, Hilden, Germany) and 5 .mu.g of plasmid DNA according
to the manufacturer's protocol. After cultivation for 24 hours
cells were washed once with PBS and cultivated again in RPMI 1640
with stabilized glutamine and 1% penicillin/streptomycin. Thus the
cell culture medium did not contain nucleosides and thereby
selection was applied on the transfected cells. Approximately 14
days after transfection the outgrowth of resistant cells is
observed. After an additional 7 to 14 days the transfectants were
tested for expression of the recombinant construct via FACS.
2.5.times.10.sup.5 cells were incubated with 50 .mu.l of an
anti-Flag-M2 antibody (Sigma-Aldrich Chemie GmbH, Taufkirchen,
Germany) diluted to 5 .mu.g/ml in PBS with 2% FCS. Bound antibody
was detected with a R-Phycoerythrin-conjugated affinity purified
F(ab')2 fragment, goat anti-mouse IgG, Fc-gamma fragment specific,
diluted 1:100 in PBS with 2% FCS (Jackson ImmunoResearch Europe
Ltd., Newmarket, Suffolk, UK). Samples were measured on a
FACScalibur (BD biosciences, Heidelberg, Germany).
[0434] 9. Generation and Characterisation of MCSP and CD3
Cross-Species Specific Bispecific Single Chain Molecules
[0435] Bispecific single chain antibody molecules each comprising a
binding domain cross-species specific for human and non-chimpanzee
primate CD3 epsilon as well as a binding domain
cross-species-specific for human and non-chimpanzee primate MCSP,
are designed as set out in the following Table 1:
TABLE-US-00004 TABLE 1 Formats of MCSP and CD3 cross-species
specific bispecific single chain antibodies SEQ ID Formats of
protein constructs (nucl/prot) (N .fwdarw. C) 190/189 MCSP-G4 HL
.times. H2C HL 192/191 MCSP-G4 HL .times. F12Q HL 194/193 MCSP-G4
HL .times. I2C HL 196/195 MCSP-G4 HLP .times. F6A HLP 198/197
MCSP-G4 HLP .times. H2C HLP 202/201 MCSP-G4 HLP .times. G4H HLP
206/205 MCSP-G4 HLP .times. E1L HLP 208/207 MCSP-G4 HLP .times. E2M
HLP 212/211 MCSP-G4 HLP .times. F12Q HL 214/213 MCSP-G4 HLP .times.
I2C HL 216/215 MCSP-D2 HL .times. H2C HL 218/217 MCSP-D2 HL .times.
F12Q HL 220/219 MCSP-D2 HL .times. I2C HL 222/221 MCSP-D2 HLP
.times. H2C HLP 224/223 MCSP-F9 HL .times. H2C HL 226/225 MCSP-F9
HLP .times. H2C HLP 228/227 MCSP-F9 HLP .times. G4H HLP 318/317
MCSP-A9 HL .times. H2C HL 320/319 MCSP-A9 HL .times. F12Q HL
322/321 MCSP-A9 HL .times. I2C HL 324/323 MCSP-C8 HL .times. I2C HL
328/327 MCSP-B7 HL .times. I2C HL 326/325 MCSP-B8 HL .times. I2C HL
330/329 MCSP-G8 HL .times. I2C HL 332/331 MCSP-D5 HL .times. I2C HL
334/333 MCSP-F7 HL .times. I2C HL 336/335 MCSP-G5 HL .times. I2C HL
338/337 MCSP-F8 HL .times. I2C HL 340/339 MCSP-G10 HL .times. I2C
HL
[0436] The aforementioned constructs containing the variable
heavy-chain (VH) and variable light-chain (VL) domains
cross-species specific for human and macaque MCSP D3 and the VH and
VL domains cross-species specific for human and macaque CD3 were
obtained by gene synthesis. The gene synthesis fragments were
designed as to contain first a Kozak site for eukaryotic expression
of the construct, followed by a 19 amino acid immunoglobulin leader
peptide, followed in frame by the coding sequence of the respective
bispecific single chain antibody molecule, followed in frame by the
coding sequence of a histidine.sub.6-tag and a stop codon. The gene
synthesis fragment was also designed as to introduce suitable N-
and C-terminal restriction sites. The gene synthesis fragment was
cloned via these restriction sites into a plasmid designated
pEF-DHFR (pEF-DHFR is described in Raum et al. Cancer Immunol
Immunother 50 (2001) 141-150) according to standard protocols
(Sambrook, Molecular Cloning; A Laboratory Manual, 3rd edition,
Cold Spring Harbour Laboratory Press, Cold Spring Harbour, N.Y.
(2001)). The constructs were transfected stably or transiently into
DHFR-deficient CHO-cells (ATCC No. CRL 9096) by electroporation or
alternatively into HEK 293 (human embryonal kidney cells, ATCC
Number: CRL-1573) in a transient manner according to standard
protocols.
[0437] Eukaryotic protein expression in DHFR deficient CHO cells
(ATCC No. CRL 9096) was performed as described by Kaufmann R. J.
(1990) Methods Enzymol. 185, 537-566. Gene amplification of the
constructs was induced by addition of increasing concentrations of
methothrexate (MTX) up to final concentrations of 20 nM MTX. After
two passages of stationary culture the cells were grown in roller
bottles with nucleoside-free HyQ PF CHO liquid soy medium (with 4.0
mM L-Glutamine with 0.1% Pluronic F-68; HyClone) for 7 days before
harvest. The cells were removed by centrifugation and the
supernatant containing the expressed protein is stored at
-20.degree. C.
[0438] Akta.RTM. Explorer System (GE Health Systems) and
Unicorn.RTM. Software were used for chromatography. Immobilized
metal affinity chromatography ("IMAC") was performed using a
Fractogel EMD chelate.RTM. (Merck) which was loaded with ZnCl.sub.2
according to the protocol provided by the manufacturer. The column
was equilibrated with buffer A (20 mM sodium phosphate buffer pH
7.2, 0.1 M NaCl) and the cell culture supernatant (500 ml) was
applied to the column (10 ml) at a flow rate of 3 ml/min. The
column was washed with buffer A to remove unbound sample. Bound
protein was eluted using a two step gradient of buffer B (20 mM
sodium phosphate buffer pH 7.2, 0.1 M NaCl, 0.5 M Imidazole)
according to the following:
[0439] Step 1: 20% buffer B in 6 column volumes
[0440] Step 2: 100% buffer B in 6 column volumes
[0441] Eluted protein fractions from step 2 were pooled for further
purification. All chemicals are of research grade and purchased
from Sigma (Deisenhofen) or Merck (Darmstadt).
[0442] Gel filtration chromatography was performed on a HiLoad
16/60 Superdex 200 prep grade column (GE/Amersham) equilibrated
with Equi-buffer (25 mM Citrate, 200 mM Lysine, 5% Glycerol, pH
7.2). Eluted protein samples (flow rate 1 ml/min) were subjected to
standard SDS-PAGE and Western Blot for detection. Prior to
purification, the column was calibrated for molecular weight
determination (molecular weight marker kit, Sigma MW GF-200).
Protein concentrations were determined using OD280 nm.
[0443] Purified bispecific single chain antibody protein was
analyzed in SDS PAGE under reducing conditions performed with
pre-cast 4-12% Bis Tris gels (Invitrogen). Sample preparation and
application were performed according to the protocol provided by
the manufacturer. The molecular weight was determined with
MultiMark protein standard (Invitrogen). The gel was stained with
colloidal Coomassie (Invitrogen protocol). The purity of the
isolated protein is >95% as determined by SDS-PAGE.
[0444] The bispecific single chain antibody has a molecular weight
of about 52 kDa under native conditions as determined by gel
filtration in phosphate buffered saline (PBS). All constructs were
purified according to this method.
[0445] Western Blot was performed using an Optitran.RTM. BA-S83
membrane and the Invitrogen Blot Module according to the protocol
provided by the manufacturer. For detection of the bispecific
single chain antibody protein antibodies an anti-His Tag antibody
was used (Penta His, Qiagen). A Goat-anti-mouse Ig antibody labeled
with alkaline phosphatase (AP) (Sigma) was used as secondary
antibody and BCIP/NBT (Sigma) as substrate. A single band was
detected at 52 kD corresponding to the purified bispecific single
chain antibody.
[0446] Alternatively, constructs were transiently expressed in DHFR
deficient CHO cells. In brief, 4.times.105 cells per construct were
cultivated in 3 ml RPMI 1640 all medium with stabilized glutamine
supplemented with 10% fetal calf serum, 1% penicillin/streptomycin
and nucleosides from a stock solution of cell culture grade
reagents (Sigma-Aldrich Chemie GmbH, Taufkirchen, Germany) to a
final concentration of 10 .mu.g/ml Adenosine, 10 .mu.g/ml
Deoxyadenosine and 10 .mu.g/ml Thymidine, in an incubator at
37.degree. C., 95% humidity and 7% CO2 one day before transfection.
Transfection was performed with Fugene 6 Transfection Reagent
(Roche, #11815091001) according to the manufacturer's protocol. 94
.mu.l OptiMEM medium (Invitrogen) and 6 .mu.l Fugene 6 are mixed
and incubated for 5 minutes at room temperature. Subsequently, 1.5
.mu.g DNA per construct were added, mixed and incubated for 15
minutes at room temperature. Meanwhile, the DHFR deficient CHO
cells were washed with 1.times.PBS and resuspended in 1.5 ml RPMI
1640 all medium.
[0447] The transfection mix was diluted with 600 .mu.l RPMI 1640
all medium, added to the cells and incubated overnight at
37.degree. C., 95% humidity and 7% CO2. The day after transfection
the incubation volume of each approach was extended to 5 ml RPMI
1640 all medium. Supernatant was harvested after 3 days of
incubation.
[0448] 10. Flow Cytometric Binding Analysis of the MCSP and CD3
Cross-Species Specific Bispecific Antibodies
[0449] In order to test the functionality of the cross-species
specific bispecific antibody constructs regarding the capability to
bind to human and macaque MCSP D3 and CD3, respectively, a FACS
analysis was performed. For this purpose CHO cells transfected with
human MCSP D3 (as described in Example 7) and the human CD3
positive T cell leukemia cell line HPB-ALL (DSMZ, Braunschweig,
ACC483) were used to test the binding to human antigens. The
binding reactivity to macaque antigens was tested by using the
generated macaque MCSP D3 transfectant (described in Example 8) and
a macaque T cell line 4119LnPx (kindly provided by Prof.
Fickenscher, Hygiene Institute, Virology, Erlangen-Nuernberg;
published in Knappe A, et al., and Fickenscher H., Blood 2000, 95,
3256-61). 200,000 cells of the respective cell lines were incubated
for 30 min on ice with 50 .mu.l of the purified protein of the
cross-species specific bispecific antibody constructs (2 .mu.g/ml)
or cell culture supernatant of transfected cells expressing the
cross-species specific bispecific antibody constructs. The cells
were washed twice in PBS with 2% FCS and binding of the construct
was detected with a murine anti-His antibody (Penta His antibody;
Qiagen; diluted 1:20 in 50 .mu.l PBS with 2% FCS). After washing,
bound anti-His antibodies were detected with an Fc gamma-specific
antibody (Dianova) conjugated to phycoerythrin, diluted 1:100 in
PBS with 2% FCS. Supernatant of untransfected CHO cells was used as
negative control for binding to the T cell lines. A single chain
construct with irrelevant target specificity was used as negative
control for binding to the MCSP-D3 transfected CHO cells.
[0450] Flow cytometry was performed on a FACS-Calibur apparatus;
the CellQuest software was used to acquire and analyze the data
(Becton Dickinson biosciences, Heidelberg). FACS staining and
measuring of the fluorescence intensity were performed as described
in Current Protocols in Immunology (Coligan, Kruisbeek, Margulies,
Shevach and Strober, Wiley-Interscience, 2002).
[0451] The bispecific binding of the single chain molecules listed
above, which are cross-species specific for MCSP D3 and
cross-species specific for human and macaque CD3 was clearly
detectable as shown in FIGS. 10, 11, 12 and 39A-39L. In the FACS
analysis all constructs showed binding to CD3 and MCSP D3 as
compared to the respective negative controls. Cross-species
specificity of the bispecific antibodies to human and macaque CD3
and MCSP D3 antigens was demonstrated.
[0452] 11. Bioactivity of MCSP and CD3 Cross-Species Specific
Bispecific Single Chain Antibodies
[0453] Bioactivity of the generated bispecific single chain
antibodies was analyzed by chromium 51 (.sup.51Cr) release in vitro
cytotoxicity assays using the MCSP D3 positive cell lines described
in Examples 7 and 8. As effector cells stimulated human CD4/CD56
depleted PBMC, stimulated human PBMC or the macaque T cell line
4119LnPx are used as specified in the respective figures.
[0454] Generation of the stimulated CD4/CD56 depleted PBMC was
performed as follows:
[0455] Coating of a Petri dish (145 mm diameter, Greiner bio-one
GmbH, Kremsmunster) was carried out with a commercially available
anti-CD3 specific antibody (e.g. OKT3, Othoclone) in a final
concentration of 1 .mu.g/ml for 1 hour at 37.degree. C. Unbound
protein was removed by one washing step with PBS. The fresh PBMC
were isolated from peripheral blood (30-50 ml human blood) by
Ficoll gradient centrifugation according to standard protocols.
3-5.times.10.sup.7 PBMC were added to the precoated petri dish in
120 ml of RPMI 1640 with stabilized glutamine/10% FCS/IL-2 20 U/ml
(Proleukin, Chiron) and stimulated for 2 days. On the third day the
cells were collected and washed once with RPMI 1640. IL-2 was added
to a final concentration of 20 U/ml and the cells were cultivated
again for one day in the same cell culture medium as above. By
depletion of CD4+ T cells and CD56+ NK cells according to standard
protocols CD8+ cytotoxic T lymphocytes (CTLs) were enriched.
[0456] Target cells were washed twice with PBS and labelled with
11.1 MBq .sup.51Cr in a final volume of 100 .mu.l RPMI with 50% FCS
for 45 minutes at 37.degree. C. Subsequently the labelled target
cells were washed 3 times with 5 ml RPMI and then used in the
cytotoxicity assay. The assay was performed in a 96 well plate in a
total volume of 250 .mu.l supplemented RPMI (as above) with an E:T
ratio 10:1. 1 .mu.g/ml of the cross-species specific bispecific
single chain antibody molecules and 20 threefold dilutions thereof
were applied. If using supernatant containing the cross-species
specific bispecific single chain antibody molecules, 21 two- and 20
threefold dilutions thereof were applied for the macaque and the
human cytotoxicity assay, respectively. The assay time was 18 hours
and cytotoxicity was measured as relative values of released
chromium in the supernatant related to the difference of maximum
lysis (addition of Triton-X) and spontaneous lysis (without
effector cells). All measurements were done in quadruplicates.
Measurement of chromium activity in the supernatants was performed
with a Wizard 3'' gamma counter (Perkin Elmer Life Sciences GmbH,
Koln, Germany). Analysis of the experimental data was performed
with Prism 4 for Windows (version 4.02, GraphPad Software Inc., San
Diego, Calif., USA). Sigmoidal dose response curves typically have
R.sup.2 values >0.90 as determined by the software. EC.sub.50
values calculated by the analysis program were used for comparison
of bioactivity.
[0457] As shown in FIGS. 13A-13B, 14A-14B, 15A-15B, 16A-16B,
17A-17B and 40A-40D, all of the generated cross-species specific
bispecific single chain antibody constructs demonstrate cytotoxic
activity against human MCSP D3 positive target cells elicited by
stimulated human CD4/CD56 depleted PBMC or stimulated PBMC and
against macaque MCSP D3 positive target cells elicited by the
macaque T cell line 4119LnPx.
[0458] 12. Plasma Stability of MCSP and CD3 Cross-Species Specific
Bispecific Single Chain Antibodies
[0459] Stability of the generated bispecific single chain
antibodies in human plasma was analyzed by incubation of the
bispecific single chain antibodies in 50% human Plasma at
37.degree. C. and 4.degree. C. for 24 hours and subsequent testing
of bioactivity. Bioactivity was studied in a chromium 51
(.sup.51Cr) release in vitro cytotoxicity assay using a MCSP
positive CHO cell line (expressing MCSP as cloned according to
example 14 or 15) as target and stimulated human CD8 positive T
cells as effector cells.
[0460] EC.sub.50 values calculated by the analysis program as
described above were used for comparison of bioactivity of
bispecific single chain antibodies incubated with 50% human plasma
for 24 hours at 37.degree. C. and 4.degree. C. respectively with
bispecific single chain antibodies without addition of plasma or
mixed with the same amount of plasma immediately prior to the
assay.
[0461] As shown in FIGS. 18(1)-18(3) and Table 2 the bioactivity of
the G4 H-L.times.I2C H-L, G4 H-L.times.H2C H-L and G4
H-L.times.F12Q H-L bispecific antibodies was not significantly
reduced as compared with the controls without the addition of
plasma or with addition of plasma immediately before testing of
bioactivity.
TABLE-US-00005 TABLE 2 bioactivity of the bispecific antibodies
without or with the addition of Plasma Without With Plasma Plasma
Construct plasma plasma 37.degree. C. 4.degree. C. G4 H-L .times.
I2C H-L 300 796 902 867 G4 H-L .times. H2C H-L 496 575 2363 1449 G4
H-L .times. F12Q H-L 493 358 1521 1040
[0462] 13. Redistribution of Circulating T Cells in the Absence of
Circulating Target Cells by First Exposure to CD3 Binding Molecules
Directed at Conventional i.e. Context Dependent CD3 Epitopes is a
Major Risk Factor for Adverse Events Related to the Initiation of
Treatment
[0463] T Cell Redistribution in Patients with B-Cell
Non-Hodgkin-Lymphoma (B-NHL) Following Initiation of Treatment with
the Conventional CD3 Binding Molecule
[0464] A conventional CD19.times.CD3 binding molecules is a CD3
binding molecule of the bispecific tandem scFv format (Loffler
(2000, Blood, Volume 95, Number 6) or WO 99/54440). It consists of
two different binding portions directed at (i) CD19 on the surface
of normal and malignant human B cells and (ii) CD3 on human T
cells. By crosslinking CD3 on T cells with CD19 on B cells, this
construct triggers the redirected lysis of normal and malignant B
cells by the cytotoxic activity of T cells. The CD3 epitope
recognized by such a conventional CD3 binding molecule is localized
on the CD3 epsilon chain, where it only takes the correct
conformation if it is embedded within the rest of the epsilon chain
and held in the right position by heterodimerization of the epsilon
chain with either the CD3 gamma or delta chain. Interaction of this
highly context dependent epitope with a conventional CD3 binding
molecule (see e.g. Loffler (2000, Blood, Volume 95, Number 6) or WO
99/54440)--even when it occurs in a purely monovalent fashion and
without any crosslinking--can induce an allosteric change in the
conformation of CD3 leading to the exposure of an otherwise hidden
proline-rich region within the cytoplasmic domain of CD3 epsilon.
Once exposed, the proline-rich region can recruit the signal
transduction molecule Nck2, which is capable of triggering further
intracellular signals. Although this is not sufficient for full T
cell activation, which definitely requires crosslinking of several
CD3 molecules on the T cell surface, e.g. by crosslinking of
several anti-CD3 molecules bound to several CD3 molecules on a T
cell by several CD19 molecules on the surface of a B cell, pure
monovalent interaction of conventional CD3 binding molecules to
their context dependent epitope on CD3 epsilon is still not inert
for T cells in terms of signalling. Without being bound by theory,
monovalent conventional CD3 binding molecules (known in the art)
may induce some T cell reactions when infused into humans even in
those cases where no circulating target cells are available for CD3
crosslinking. An important T cell reaction to the intravenous
infusion of monovalent conventional CD19.times.CD3 binding molecule
into B-NHL patients who have essentially no circulating
CD19-positive B cells is the redistribution of T cells after start
of treatment. It has been found in a phase I clinical trial that
this T cell reaction occurs during the starting phase of
intravenous CD19.times.CD3 binding molecule infusion in all
individuals without circulating CD19-positive target B cells
essentially independent of the CD19.times.CD3 binding molecule dose
(FIG. 19). However, sudden increases in CD19.times.CD3 binding
molecule exposure have been found to trigger virtually the same
redistributional T cell reaction in these patients as the initial
exposure of T cells to CD19.times.CD3 binding molecule at treatment
start (FIG. 20A) and even gradual increases in CD19.times.CD3
binding molecule exposure still can have redistributional effects
on circulating T cells (FIG. 21). Moreover, it has been found that
this essentially dose-independent redistributional T cell reaction
in the absence of circulating target cells as triggered by
conventional CD3 binding molecules like the CD19.times.CD3 binding
molecule (e.g. disclosed in WO 99/54440) in 100% of all treated
individuals is a major risk factor for adverse events related to
the initiation of treatment.
[0465] According to the study protocol, patients with relapsed
histologically confirmed indolent B-cell Non-Hodgkin-Lymphoma
(B-NHL) including mantle cell lymphoma were recruited in an
open-label, multi-center phase I interpatient dose-escalation
trial. The study protocol was approved by the independent ethics
committees of all participating centers and sent for notification
to the responsible regulatory authority. Measurable disease (at
least one lesion .gtoreq.1.5 cm) as documented by CT scan was
required for inclusion into the study. Patients received
conventional CD19.times.CD3 binding molecule by continuous
intravenous infusion with a portable minipump system over four
weeks at constant flow rate (i.e. dose level). Patients were
hospitalized during the first two weeks of treatment before they
were released from the hospital and continued treatment at home.
Patients without evidence of disease progression after four weeks
were offered to continue treatment for further four weeks. So far
six different dose levels were tested without reaching a maximum
tolerated dose (MTD): 0.5, 1.5, 5, 15, 30 and 60 .mu.g/m.sup.2/24
h. Cohorts consisted of three patients each if no adverse events
defined by the study protocol as DLT (dose limiting toxicity) were
observed. In case of one DLT among the first three patients the
cohort was expanded to six patients, which--in the absence of a
second DLT--allowed further dose escalation. Accordingly, dose
levels without DLT in cohorts with 3 patients or with one DLT in
cohorts with 6 patients were regarded as safe. Study treatment was
stopped in all patients who developed a DLT. At 15 and 30
.mu.g/m.sup.2/24 h different modes of treatment initiation during
the first 24 h were tested in several additional cohorts: (i)
Stepwise increase after 5 .mu.g/m.sup.2/24 h for the first 24 h to
15 .mu.g/m.sup.2/24 h maintenance dose (patient cohort 15-step),
(ii) even continuous increase of flow-rate from almost zero to 15
or 30 .mu.g/m.sup.2/24 h (patient cohorts 15-ramp and 30-ramp) and
(iii) start with the maintenance dose from the very beginning
(patient cohorts 15-flat, 30-flat and 60-flat). Patient cohorts at
dose levels 0.5, 1.5 and 5 .mu.g/m.sup.2/24 h were all started with
the maintenance dose from the very beginning (i.e. flat
initiation).
[0466] Time courses of absolute B- and T-cell counts in peripheral
blood were determined by four color FACS analysis as follows:
[0467] Collection of Blood Samples and Routine Analysis
[0468] In patient cohorts 15-ramp, 15-flat, 30-ramp, 30-flat and
60-flat blood samples (6 ml) were obtained before and 0.75, 2, 6,
12, 24, 30, 48 hours after start of CD19.times.CD3 binding molecule
(as disclosed in WO 99/54440) infusion as well as on treatment days
8, 15, 17, 22, 24, 29, 36, 43, 50, 57 and 4 weeks after end of
conventional CD19.times.CD3 binding molecule infusion using
EDTA-containing Vacutainer.TM. tubes (Becton Dickinson) which were
shipped for analysis at 4.degree. C. In patient cohorts 15-step
blood samples (6 ml) were obtained before and 6, 24, 30, 48 hours
after start of CD19.times.CD3 binding molecule infusion as well as
on treatment days 8, 15, 22, 29, 36, 43, 50, 57 and 4 weeks after
end of CD19.times.CD3 binding molecule infusion. At dose levels
0.5, 1.5 and 5 .mu.g/m.sup.2/24 h blood samples (6 ml) were
obtained before and 6, 24, 48 hours after start of CD19.times.CD3
binding molecule infusion as well as on treatment days 8, 15, 22,
29, 36, 43, 50, 57 and 4 weeks after end of CD19.times.CD3 binding
molecule infusion. In some cases slight variations of these time
points occurred for operational reasons. FACS analysis of
lymphocyte subpopulations was performed within 24-48 h after blood
sample collection. Absolute numbers of leukocyte subpopulations in
the blood samples were determined through differential blood
analysis on a CoulterCounter.TM. (Coulter).
[0469] Isolation of PBMC from Blood Samples
[0470] PBMC (peripheral blood mononuclear cells) isolation was
performed by an adapted Ficoll.TM. gradient separation protocol.
Blood was transferred at room temperature into 10 ml Leucosep.TM.
tubes (Greiner) pre-loaded with 3 ml Biocoll.TM. solution
(Biochrom). Centrifugation was carried out in a swing-out rotor for
15 min at 1700.times.g and 22.degree. C. without deceleration. The
PBMC above the Biocoll.TM. layer were isolated, washed once with
FACS buffer (PBS/2% FBS [Foetal Bovine Serum; Biochrom]),
centrifuged and resuspended in FACS buffer. Centrifugation during
all wash steps was carried out in a swing-out rotor for 4 min at
800.times.g and 4.degree. C. If necessary, lysis of erythrocytes
was performed by incubating the isolated PBMC in 3 ml erythrocyte
lysis buffer (8.29 g NH.sub.4Cl, 1.00 g KHCO.sub.3, 0.037 g EDTA,
ad 1.0 I H.sub.2O.sub.bidest, pH 7.5) for 5 min at room temperature
followed by a washing step with FACS buffer.
[0471] Staining of PBMC with Fluorescence-Labeled Antibodies
Against Cell Surface Molecules
[0472] Monoclonal antibodies were obtained from Invitrogen
(.sup.1Cat. No. MHCD1301, .sup.2Cat. No. MHCD1401), Dako
(.sup.5Cat. No. C7224) or Becton Dickinson (.sup.3Cat. No. 555516,
.sup.4Cat. No. 345766) used according to the manufacturers'
recommendations. 5.times.10.sup.5-1.times.10.sup.6 cells were
stained with the following antibody combination:
anti-CD13.sup.1/anti-CD14.sup.2 (FITC).times.anti-CD56.sup.3
(PE).times.anti-CD3.sup.4 (PerCP).times.anti-CD19.sup.5 (APC).
Cells were pelleted in V-shaped 96 well multititer plates (Greiner)
and the supernatant was removed. Cell pellets were resuspended in a
total volume of 100 .mu.l containing the specific antibodies
diluted in FACS buffer. Incubation was carried out in the dark for
30 min at 4.degree. C. Subsequently, samples were washed twice with
FACS buffer and cell pellets were resuspended in FACS buffer for
flowcytometric analysis.
[0473] Flowcytometric Detection of Stained Lymphocytes by FACS
[0474] Data collection was performed with a 4 color BD
FACSCalibur.TM. (Becton Dickinson). For each measurement
1.times.10.sup.4 cells of defined lymphocyte subpopulations were
acquired. Statistical analysis was performed with the program
CellQuest Pro.TM. (Becton Dickinson) to obtain lymphocyte
subpopulation percentages and to classify cell surface molecule
expression intensity. Subsequently, percentages of single
lymphocyte subsets related to total lymphocytes (i.e. B plus T plus
NK cells excluding any myeloid cells via CD13/14-staining) as
determined by FACS were correlated with the lymphocyte count from
the differential blood analysis to calculate absolute cell numbers
of T cells (CD3.sup.+, CD56.sup.-, CD13/14.sup.-) and B cells
(CD19.sup.+, CD13/14.sup.-).
[0475] T cell redistribution during the starting phase of
conventional CD19.times.CD3 binding molecule (e.g. disclosed in WO
99/54440) treatment in all those patients who had essentially no
circulating CD19-positive B cells at treatment start is shown in
(FIG. 19). For comparison, a representative example of T cell
redistribution during the starting phase of CD19.times.CD3 binding
molecule treatment in a patient with a significant number of
circulating CD19-positive B cells is shown in FIG. 22.
[0476] In both cases (i.e. essentially no or many circulating B
cells) circulating T cell counts rapidly decrease upon treatment
start. However, in the absence of circulating B cells T cells tend
to return into the circulating blood very early, while the return
of T cells into the circulating blood of those patients who have a
significant number of circulating B cells at treatment start is
usually delayed until these circulating B cells are depleted. Thus,
the T cell redistribution patterns mainly differ in the kinetics of
T cell reappearance in the circulating blood.
[0477] Assessment of efficacy based on CT scan was carried out by
central reference radiology after 4 weeks of treatment and in
patients receiving additional 4 weeks also after 8 weeks of
treatment plus in all cases four weeks after end of treatment.
Disappearance and/or normalization in size of all known lesions
(including an enlarged spleen) plus clearance of bone marrow from
lymphoma cells in cases of bone marrow infiltration was counted as
complete response (CR). Reduction by at least 50% from baseline of
the sum of products of the two biggest diameters (SPD) of each
predefined target lesion was defined as partial response (PR); a
reduction by at least 25% was regarded a minimal response (MR).
Progressive disease (PD) was defined as 50% increase of SPD from
baseline. SPD deviations from baseline between +50% and -25% were
regarded as stable disease (SD).
[0478] Patient demographics, doses received and clinical outcome in
34 patients are summarized in Table 3. Clinical anti-tumor activity
of the CD19.times.CD3 binding molecule was clearly dose dependent:
Consistent depletion of circulating CD19-positive B (lymphoma) cell
from peripheral blood was observed from 5 .mu.g/m.sup.2/24 h
onwards. At 15 .mu.g/m.sup.2/24 h and 30 .mu.g/m.sup.2/24 h first
objective clinical responses (PRs and CRs) were recorded as well as
cases of partial and complete elimination of B lymphoma cells from
infiltrated bone marrow. Finally, at 60 .mu.g/m.sup.2/24 h the
response rate increased to 100% (PRs and CRs) and bone marrow
clearance from B lymphoma cells was complete in all evaluable
cases.
[0479] The CD19.times.CD3 binding molecule was well tolerated by
the majority of patients. Most frequent adverse events of grades
1-4 in 34 patients, regardless of causality are summarized in Table
4. CD19.times.CD3 binding molecule-related adverse events usually
were transient and fully reversible. In particular, there were 2
patients (patients #19 and #24 in Table 3) essentially without
circulating CD19-positive B cells whose treatment was stopped early
because of CNS adverse events (lead symptoms: confusion and
disorientation) related to repeated T cell redistribution during
the starting phase of CD19.times.CD3 binding molecule infusion.
[0480] One of these patients (#19) was in cohort 15-step. He
received 5 .mu.g/m.sup.2/24 h CD19.times.CD3 binding molecule for
the first 24 h followed by sudden increase to 15 .mu.g/m.sup.2/24 h
maintenance dose. The corresponding T cell redistribution pattern
shows that circulating T cell counts rapidly decreased upon start
of infusion at 5 .mu.g/m.sup.2/24 h followed by early reappearance
of T cells in the circulating blood essentially without circulating
CD19-positive B cells. As a consequence, the peripheral T cell
counts had fully recovered when the CD19.times.CD3 binding molecule
dose was increased after 24 h from 5 to 15 .mu.g/m.sup.2/24 h.
Therefore the dose step could trigger a second episode of T cell
redistribution as shown in FIG. 20A. This repeated T cell
redistribution was related with CNS side effects (lead symptoms:
confusion and disorientation) in this patient, which led to the
stop of infusion. The relationship between repeated T cell
redistribution and such CNS adverse events was also observed in
previous phase I clinical trials in B-NHL patients who received
CD19.times.CD3 binding molecule (e.g. disclosed in WO 99/54440) as
repeated bolus infusion for 2 to 4 hours each usually followed by 2
days of treatment free interval (FIG. 20B). Every single bolus
infusion triggered one episode of T cell redistribution consisting
of a fast decrease in circulating T cell counts and T cell recovery
prior to the next bolus infusion. In total, CNS adverse events
related to repeated T cell redistribution were observed in 5 out of
21 patients. FIG. 20B shows the representative example of one
patient from the bolus infusion trials, who developed CNS symptoms
after the third episode of T cell redistribution. Typically,
patients with CNS adverse events in the bolus infusion trials also
had low circulating B cell counts.
[0481] The second patient (#24) from the continuous infusion trial,
whose treatment was stopped early because of CNS adverse events
(lead symptoms: confusion and disorientation) related to repeated T
cell redistribution during the starting phase of CD19.times.CD3
binding molecule infusion, was in cohort 15-flat. By mistake, this
patient received an CD19.times.CD3 binding molecule infusion
without additional HSA as required for stabilization of the drug.
The resulting uneven drug flow triggered repeated episodes of T
cell redistribution instead of only one (FIG. 23A) with the
consequence that the infusion had to be stopped because of
developing CNS symptoms. Yet, when the same patient was restarted
correctly with CD19.times.CD3 binding molecule solution containing
additional HSA for drug stabilization (e.g. disclosed in WO
99/54440), no repeated T cell redistribution was observed and the
patient did not again develop any CNS symptoms (FIG. 23B). Because
this patient also had essentially no circulating B cells, the
circulating T cells could react with fast redistribution kinetics
even to subtle changes in drug exposure as observed. The CNS
adverse events related to T cell redistribution in patients who
have essentially no circulating target cells can be explained by a
transient increase of T cell adhesiveness to the endothelial cells
followed by massive simultaneous adhesion of circulating T cells to
the blood vessel walls with a consecutive drop of T cell numbers in
the circulating blood as observed. The massive simultaneous
attachment of T cells to the blood vessel walls can cause an
increase in endothelial permeability and endothelial cell
activation. The consequences of increased endothelial permeability
are fluid shifts from the intravascular compartment into
interstitial tissue compartments including the CNS interstitium.
Endothelial cell activation by attached T cells can have
procoagulatory effects (Monaco et al. J Leukoc Biol 71 (2002)
659-668) with possible disturbances in blood flow (including
cerebral blood flow) particularly with regard to capillary
microcirculation. Thus, CNS adverse events related to T cell
redistribution in patients essentially without circulating target
cells can be the consequence of capillary leak and/or disturbances
in capillary microcirculation through adherence of T cells to
endothelial cells. The endothelial stress caused by one episode of
T cell redistribution is tolerated by the majority of patients,
while the enhanced endothelial stress caused by repeated T cell
redistribution frequently causes CNS adverse events. More than one
episode of T cell redistribution may be less risky only in patients
who have low baseline counts of circulating T cells. However, also
the limited endothelial stress caused by one episode of T cell
redistribution can cause CNS adverse events in rare cases of
increased susceptibility for such events as observed in 1 out of 21
patients in the bolus infusion trials with the CD19.times.CD3
binding molecule.
[0482] Without being bound by theory, the transient increase of T
cell adhesiveness to the endothelial cells in patients who have
essentially no circulating target cells can be explained as T cell
reaction to the monovalent interaction of a conventional CD3
binding molecule, like the CD19.times.CD3 binding molecule (e.g. WO
99/54440), to its context dependent epitope on CD3 epsilon
resulting in an allosteric change in the conformation of CD3
followed by the recruitment of Nck2 to the cytoplasmic domain of
CD3 epsilon as described above. As Nck2 is directly linked to
integrins via PINCH and ILK (FIG. 28), recruitment of Nck2 to the
cytoplasmic domain of CD3 epsilon following an allosteric change in
the conformation of CD3 through binding of a conventional CD3
binding molecule, like the CD19.times.CD3 binding molecule, to its
context dependent epitope on CD3 epsilon, can increase the
adhesiveness of T cells to endothelial cells by transiently
switching integrins on the T cell surface into their more adhesive
isoform via inside-out-signalling.
TABLE-US-00006 TABLE 3 Patient demographics and clinical outcome
Best Response* Disease (CR Duration Age/ (Ann Arbor Dose Level
Clearance of in Months or Cohort Patient Sex Classification)
[mg/m.sup.2/Day] Bone Marrow Weeks) 1 1 71/m IC, Binet C 0.0005
None SD 2 67/f MCL, Stage 0.0005 n.d. PD IV/A/E 3 67/m CLL, Stage
0.0005 n.d. MR IV/B/E 2 4 69/m MCL, Stage 0.0015 n.i. SD IV/B 5
49/m MCL, Stage 0.0015 n.d. SD IV/A/S 6 71/m MCL, Stage 0.0015 n.i.
PD IV/B/E 7 77/m MCL, Stage 0.0015 n.i. SD IV/B/E/S 8 65/m CLL,
Stage 0.0015 n.d. PD IV/B/E/S 9 75/m FL, Stage II/B 0.0015 n.i. SD
3 10 58/m MCL, Stage 0.005 n.i. PD III/B/S 11 68/f FL, Stage IV/B
0.005 n.d. SD 12 65/m MCL, Stage 0.005 n.i. SD III/A/E 4.sup.a 13
60/m SLL, Stage 0.015 Complete PR IV/B/S 14 73/m MCL, Stage 0.015
n.i. SD II/A/E 15 44/m FL, Stage 0.015 Partial PR IV/B/E/S 16 61/m
FL, Stage 0.015 Complete CR (7 mo) IV/A/S 17 67/m MZL, Stage 0.015
n.i. n.e. IV/B/S 18 64/m FL, Stage 0.015 n.i. PD IV/A/E 19 75/m
MCL, Stage 0.015 n.i. n.e. III/A 20 65/f FL; Stage III/A 0.015 n.i.
SD 21 60/m MCL, Stage 0.015 None SD IV/A/E 22 67/f FL, Stage IV/B
0.015 Complete MR 23 67/m DLBCL, Stage 0.015 n.i. n.e. III/B 24
65/f FL, Stage III/A 0.015 n.d. SD 25 74/f WD, Stage IV/B 0.015
Partial SD 5 26 67/m MCL, Stage 0.03 Complete SD IV/A 27 48/m FL,
Stage III/A 0.03 n.i. PD 28 58/m MCL, Stage 0.03 n.i. CR (10 mo+)
III/A 29 45/f MCL, Stage 0.03 Partial PD IV/B 30 59/m MZL, Stage
0.03 n.i. n.e. III/A 31 43/m FL, Stage III/A 0.03 n.i. MR 6 32 72/m
MCL, Stage 0.06 Complete PR IV/A 33 55/m MCL, Stage 0.06 Complete
CR (4 mo+) IV/B 34 52/m FL, Stage IV/A 0.06 n.i. CR.sup.b (1 w+)
*Centrally confirmed complete (CR) and partial (PR) responses by
Cheson criteria in bold; MR, minimal response (.gtoreq.25 to
<50%); SD, stable disease; PD, progressive disease; duration
from first documentation of response in parentheses; + denotes an
ongoing response .sup.aCohort 4 was expanded to study three
different schedules of treatment initiation .sup.bPR after 8 weeks
of treatment that turned into a CR after an additional treatment
cycle of 4 weeks at the same dose following 7 weeks of treatment
free interval n.e.: not evaluable, because of treatment period <
7 d n.d.: not determined (infiltrated, but no second biopsy
performed at end of treatment) n.i.: not infiltrated at start of
treatment
TABLE-US-00007 TABLE 4 Incidence of adverse events observed during
treatment Adverse events regardless of relationship, occuring in
.gtoreq.3 patients Grade 1-4 Grade 3-4 (N = 34) N (%) N (%) Pyrexia
22 (64.7) 2 (5.9) Leukopenia 21 (61.8) 11 (32.4) Lymphopenia 21
(61.8) 21 (61.8) Coagulopathy (increase in D-dimers) 16 (47.1) 6
(17.6) Enzyme abnormality (AP, LDH, CRP) 16 (47.1) 10 (29.4)
Hepatic function abnormality (ALT, AST, 16 (47.1) 1 (2.9) GGT)
Anaemia 13 (38.2) 5 (14.7) Chills 13 (38.2) 0 (0.0) Headache 12
(35.3) 1 (2.9) Hypokalaemia 12 (35.3) 2 (5.9) Thrombocytopenia 12
(35.3) 6 (17.6) Weight increased 12 (35.3) 0 (0.0) Hyperglycaemia
11 (32.4) 2 (5.9) Neutropenia 11 (32.4) 8 (23.5) Haematuria 10
(29.4) 0 (0.0) Oedema peripheral 10 (29.4) 2 (5.9) Anorexia 9
(26.5) 1 (2.9) Diarrhoea 9 (26.5) 0 (0.0) Weight decreased 9 (26.5)
0 (0.0) Fatigue 8 (23.5) 1 (2.9) Proteinuria 8 (23.5) 0 (0.0)
Hypocalcaemia 7 (20.6) 2 (5.9) Pancreatic enzyme abnormality 7
(20.6) 0 (0.0) Cough 6 (17.6) 0 (0.0) Dyspnoea 6 (17.6) 0 (0.0)
Back pain 5 (14.7) 0 (0.0) Catheter site pain 5 (14.7) 0 (0.0)
Hyperbilirubinaemia 5 (14.7) 2 (5.9) Hypoalbuminaemia 5 (14.7) 0
(0.0) Hypogammaglobulinaemia 5 (14.7) 1 (2.9) Hypoproteinaemia 5
(14.7) 0 (0.0) Pleural effusion 5 (14.7) 1 (2.9) Vomiting 5 (14.7)
0 (0.0) Asthenia 4 (11.8) 1 (2.9) Confusional state 4 (11.8) 0
(0.0) Constipation 4 (11.8) 0 (0.0) Dizziness 4 (11.8) 0 (0.0)
Hypertension 4 (11.8) 0 (0.0) Hyponatraemia 4 (11.8) 2 (5.9)
Mucosal dryness 4 (11.8) 0 (0.0) Muscle spasms 4 (11.8) 0 (0.0)
Nausea 4 (11.8) 0 (0.0) Night sweats 4 (11.8) 0 (0.0) Abdominal
pain 3 (8.8) 1 (2.9) Ascites 3 (8.8) 0 (0.0) Hypercoagulation 3
(8.8) 0 (0.0) Hyperhidrosis 3 (8.8) 0 (0.0) Hypoglobulinaemia 3
(8.8) 0 (0.0) Insomnia 3 (8.8) 0 (0.0) Liver disorder 3 (8.8) 1
(2.9) Nasopharyngitis 3 (8.8) 0 (0.0) Pruritus 3 (8.8) 0 (0.0)
[0483] Abbreviations used are: AE, adverse event; AP, alkaline
phosphatase; LDH, lactate dehydrogenase; CRP, C-reactive protein;
ALT, alanine transaminase; AST, aspartate transaminase; GGT,
gamma-glutamyl transferase; AE data from the additional treatment
cycle of patient 34 not yet included.
[0484] As explained above, conventional CD3 binding molecules (e.g.
disclosed in WO 99/54440) capable of binding to a context-dependent
epitope, though functional, lead to the undesired effect of T cell
redistribution in patients causing CNS adverse events. In contrast,
binding molecules of the present invention, by binding to the
context-independent N-terminal 1-27 amino acids of the CD3 epsilon
chain, do not lead to such T cell redistribution effects. As a
consequence, the CD3 binding molecules of the invention are
associated with a better safety profile compared to conventional
CD3 binding molecules.
[0485] 14. Bispecific CD3 Binding Molecules of the Invention
Inducing T Cell Mediated Target Cell Lysis by Recognizing a Surface
Target Antigen Deplete Target Antigen Positive Cells In Vivo
[0486] A Bispecific CD3 Binding Molecule of the Invention
Recognizing CD33 as Target Antigen Depletes CD33-Positive
Circulating Monocytes from the Peripheral Blood of Cynomolgus
Monkeys
[0487] CD33-AF5 VH-VL.times.I2C VH-VL (amino acid sequence: SEQ ID
NO.267) was produced by expression in CHO cells using the coding
nucleotide sequence SEQ ID NO. 268. The coding sequences of (i) an
N-terminal immunoglobulin heavy chain leader comprising a start
codon embedded within a Kozak consensus sequence and (ii) a
C-terminal His.sub.6-tag followed by a stop codon were both
attached in frame to the nucleotide sequence SEQ ID NO 268 prior to
insertion of the resulting DNA-fragment as obtained by gene
synthesis into the multiple cloning site of the expression vector
pEF-DHFR (Raum et al. Cancer Immunol Immunother 50 (2001) 141-150).
Stable transfection of DHFR-deficient CHO cells, selection for
DHFR-positive transfectants secreting the CD3 binding molecule
CD33-AF5 VH-VL.times.I2C VH-VL into the culture supernatant and
gene amplification with methotrexat for increasing expression
levels were carried out as described (Mack et al. Proc. Natl. Acad.
Sci. USA 92 (1995) 7021-7025). The analytical SEC-profile of
CD33-AF5 VH-VL.times.I2C VH-VL for use in cynomolgus monkeys
revealed that the test material almost exclusively consisted of
monomer. The potency of the test material was measured in a
cytotoxicity assay as described in example 16.5 using CHO cells
transfected with cynomolgus CD33 as target cells and the macaque T
cell line 4119LnPx as source of effector cells (FIG. 25). The
concentration of CD33-AF5 VH-VL.times.I2C VH-VL required for
half-maximal target cell lysis by the effector T cells (EC50) was
determined to be 2.7 ng/ml. Young (approx. 3 years old) adult
cynomolgus monkeys (Macaca fascicularis) were treated by continuous
intravenous infusion of CD3 binding molecule CD33-AF5
VH-VL.times.I2C VH-VL at different flow-rates (i.e. dose levels) to
study depletion of circulating CD33-positive monocytes from the
peripheral blood. This situation is equivalent to the treatment
with the conventional CD3 binding molecule CD19.times.CD3 (specific
for CD19 on B cells and CD3 on T cells) of those B-NHL patients,
who have circulating CD19-positive target B cells (see e.g.
WO99/54440). Depletion of circulating CD19-positive target B cells
from the peripheral blood had turned out as a valid surrogate for
the general clinical efficacy of the conventional CD3 binding
molecule (CD19.times.CD3 as provided in WO99/54440) in patients
with CD19-positive B-cell malignomas like B-NHL. Likewise,
depletion of circulating CD33-positive monocytes from the
peripheral blood is regarded as a valid surrogate of the general
clinical efficacy of CD33-directed bispecific CD3 binding molecules
of the invention like CD33-AF5 VH-VL.times.I2C VH-VL in patients
with CD33-positive myeloid malignomas like AML (acute myeloid
leukemia).
[0488] Continuous infusion was carried out according to the Swivel
method as follows: The monkeys are catheterized via the vena
femoralis into the vena cava caudalis using a vein catheter. The
catheter is tunneled subcutaneously to the dorsal shoulder region
and exteriorized at the caudal scapula. Then a tube is passed
through a jacket and a protection spring. The jacket is fastened
around the animal and the catheter, via the tube, is connected to
an infusion pump.
[0489] Administration solution (1.25 M lysine, 0.1% tween 80, pH 7)
without test material was infused continuously at 48 ml/24 h for 7
days prior to treatment start to allow acclimatization of the
animals to the infusion conditions. Treatment was started by adding
CD33-AF5 VH-VL.times.I2C VH-VL test material to the administration
solution at the amount required for each individual dose level to
be tested (i.e. flow rate of CD33-AF5 VH-VL.times.I2C VH-VL). The
infusion reservoir was changed every day throughout the whole
acclimatization and treatment phase. Planned treatment duration was
7 days except for the 120 .mu.g/m.sup.2/24 h dose level, where
animals received 14 days of treatment.
[0490] Time courses of absolute counts in circulating T cells and
CD33-positive monocytes were determined by 4- or 3-colour FACS
analysis, respectively:
[0491] Collection of Blood Samples and Routine Analysis
[0492] Blood samples (1 ml) were obtained before and 0.75, 2, 6,
12, 24, 30, 48, 72 hours after start of continuous infusion with
MCSP-G4 VH-VL.times.I2C VH-VL as well as after 7 and 14 days (and
after 9 days at the 120 .mu.g/m.sup.2/24 h dose level) of treatment
using EDTA-containing Vacutainer.TM. tubes (Becton Dickinson) which
were shipped for analysis at 4.degree. C. In some cases slight
variations of these time points occurred for operational reasons.
FACS analysis of lymphocyte subpopulations was performed within
24-48 h after blood sample collection. Absolute numbers of
leukocyte subpopulations in the blood samples were determined
through differential blood analysis in a routine veterinary
lab.
[0493] Isolation of PBMC from Blood Samples
[0494] PBMC (peripheral blood mononuclear cells) were isolated in
analogy to the protocol described in example 13, above, with
adaptations of the used volumes.
[0495] Staining of PBMC with Fluorescence-Labeled Antibodies
Against Cell Surface Molecules
[0496] Monoclonal antibodies reactive with cynomolgus antigens were
obtained from Becton Dickinson (.sup.1Cat. No. 345784, .sup.2Cat.
No. 556647, .sup.3Cat. No. 552851, .sup.6Cat. No. 557710), Beckman
Coulter (.sup.4Cat. No. IM2470) and Miltenyi (.sup.5Cat. No.
130-091-732) and used according to the manufacturers'
recommendations. 5.times.10.sup.5-1.times.10.sup.6 cells were
stained with the following antibody combinations: anti-CD14.sup.1
(FITC).times.anti-CD56.sup.2 (PE).times.anti-CD3.sup.3
(PerCP).times.anti-CD19.sup.4 (APC) and anti-CD14.sup.1
(FITC).times.anti-CD33.sup.5 (PE).times.anti-CD16.sup.6 (Alexa
Fluor 647.TM.). Additional steps were performed as described in
example 13, above.
[0497] Flowcytometric Detection of Stained Lymphocytes by FACS
[0498] Data collection was performed with a 4 color BD
FACSCalibur.TM. (Becton Dickinson). For each measurement
1.times.10.sup.4 cells of defined lymphocyte subpopulations were
acquired. Statistical analysis was performed with the program
CellQuest Pro.TM. (Becton Dickinson) to obtain lymphocyte
subpopulation percentages and to classify cell surface molecule
expression intensity. Subsequently, percentages of single
lymphocyte subsets related to total lymphocytes (i.e. B plus T plus
NK cells excluding myeloid cells via CD14-staining) as determined
by FACS were correlated with the lymphocyte count from the
differential blood analysis to calculate absolute cell numbers of T
cells (CD3.sup.+, CD56.sup.-, CD14.sup.-). Absolute numbers of
CD33-positive monocytes were calculated by multiplying the monocyte
counts from the differential blood analysis with the corresponding
ratios of CD33-positive monocytes (CD33.sup.+, CD14.sup.+) to all
monocytes (CD14.sup.+) as determined by FACS.
[0499] The percentage compared to baseline (i.e. 100%) of absolute
circulating CD33-positive monocyte counts at the end of treatment
with CD33-AF5 VH-VL.times.I2C VH-VL in 4 cohorts of 2 cynomolgus
monkeys with inter-cohort dose escalation from 30 over 60 and 240
to 1000 .mu.g/m.sup.2/24 h are shown in FIG. 26A.
[0500] As shown in FIG. 26A, continuous intravenous infusion of
CD33-AF5 VH-VL.times.I2C VH-VL induces depletion of circulating
CD33-positive monocytes in a dose-dependent manner. While there was
still no detectable depletion of circulating CD33-positive
monocytes at 30 .mu.g/m.sup.2/24 h, a first trend towards a
reduction of CD33-positive monocyte counts became visible at 60
.mu.g/m.sup.2/24 h after 7 days of treatment. At 240
.mu.g/m.sup.2/24 h circulating CD33-positive monocytes were almost
completely depleted from the peripheral blood after 3 days of
treatment. This was reached even faster at 1000 .mu.g/m.sup.2/24 h,
where depletion of the circulating CD33-positive monocytes from the
peripheral blood was completed already after 1 day of treatment.
This finding was confirmed by the results shown in FIGS.
26B(1)-26B(2) demonstrating depletion of circulating CD33-positive
monocytes by two thirds and 50% compared to the respective baseline
in two cynomolgus monkeys treated by continuous infusion with
CD33-AF5 VH-VL.times.I2C VH-VL at 120 .mu.g/m.sup.2/24 h for 14
days.
[0501] This outcome is a clear signal clinical efficacy of the CD3
binding molecules of the invention in general and of bispecific
CD33-directed CD3 binding molecules of the invention for the
treatment of CD33-positive malignomas like AML in particularly.
Moreover, the T cell redistribution during the starting phase of
treatment with CD33-AF5 VH-VL.times.I2C VH-VL in the presence of
circulating target cells (i.e. CD33-positive monocytes) seems to be
less pronounced than T cell redistribution during the starting
phase of treatment with conventional CD19.times.CD3 constructs, as
described in WO99/54440 in B-NHL patients with a significant number
of circulating target cells (i.e. CD19-positive B cells) as shown
in FIG. 22. While T cells disappear completely from the circulation
upon start of CD19.times.CD3 infusion and do not reappear until the
circulating CD19-positive target B cells are depleted from the
peripheral blood (FIG. 22), initial disappearance of circulating T
cells is incomplete upon infusion start with CD33-AF5
VH-VL.times.I2C VH-VL and T cell counts recover still during the
presence of circulating CD33-positive target cells (FIG. 26 B).
This confirms that CD3 binding molecules of the invention (directed
against and generated against an epitope of human and
non-chimpanzee primates CD3.epsilon. (epsilon) chain and being a
part or fragment or the full length of the amino acid sequence as
provided in SEQ ID Nos. 2, 4, 6, or 8) by recognizing a
context-independent CD3 epitope show a more favorable T cell
redistribution profile than conventional CD3 binding molecules
recognizing a context-dependent CD3 epitope, like the binding
molecules provided in WO99/54440.
[0502] 15. CD3 Binding Molecules of the Invention Directed at
Essentially Context Independent CD3 Epitopes by Inducing Less
Redistribution of Circulating T Cells in the Absence of Circulating
Target Cells Reduce the Risk of Adverse Events Related to the
Initiation of Treatment
[0503] Reduced T Cell Redistribution in Cynomolgus Monkeys
Following Initiation of Treatment with a Representative
Cross-Species Specific CD3 Binding Molecule of the Invention
[0504] MCSP-G4 VH-VL.times.I2C VH-VL (amino acid sequence: SEQ ID
NO. 193) was produced by expression in CHO cells using the coding
nucleotide sequence SEQ ID NO. 194. The coding sequences of (i) an
N-terminal immunoglobulin heavy chain leader comprising a start
codon embedded within a Kozak consensus sequence and (ii) a
C-terminal His6-tag followed by a stop codon were both attached in
frame to the nucleotide sequence SEQ ID NO. 194 prior to insertion
of the resulting DNA-fragment as obtained by gene synthesis into
the multiple cloning site of the expression vector pEF-DHFR (Raum
et al. Cancer Immunol Immunother 50 (2001) 141-150). Stable
transfection of DHFR-deficient CHO cells, selection for
DHFR-positive transfectants secreting the CD3 binding molecule
MCSP-G4 VH-VL.times.I2C VH-VL into the culture supernatant and gene
amplification with methotrexat for increasing expression levels
were carried out as described (Mack et al. Proc. Natl. Acad. Sci.
USA 92 (1995) 7021-7025). Test material for treatment of cynomolgus
monkeys was produced in a 200-liter fermenter. Protein purification
from the harvest was based on IMAC affinity chromatography
targeting the C-terminal His6-tag of MCSP-G4 VH-VL.times.I2C VH-VL
followed by preparative size exclusion chromatography (SEC). The
total yield of final endotoxin-free test material was 40 mg. The
test material consisted of 70% monomer, 30% dimer and a small
contamination of higher multimer. The potency of the test material
was measured in a cytotoxicity assay as described in example 11
using CHO cells transfected with cynomolgus MCSP as target cells
and the macaque T cell line 4119LnPx as source of effector cells
(FIG. 27). The concentration of MCSP-G4 VH-VL.times.I2C VH-VL
required for half-maximal target cell lysis by the effector T cells
(EC50) was determined to be 1.9 ng/ml.
[0505] Young (approx. 3 years old) adult cynomolgus monkeys (Macaca
fascicularis) were treated by continuous intravenous infusion of
CD3 binding molecule MCSP-G4 VH-VL.times.I2C VH-VL at different
flow-rates (i.e. dose levels) to study redistribution of
circulating T cells following initiation of treatment in the
absence of circulating target cells. Although the CD3 binding
molecule MCSP-G4 VH-VL.times.I2C VH-VL can recognize both
cynomolgus MCSP and cynomolgus CD3, there are no circulating blood
cells expressing MCSP. Therefore, the only interaction possible in
the circulating blood is binding of the CD3-specific arm of MCSP-G4
VH-VL.times.I2C VH-VL to CD3 on T cells. This situation is
equivalent to the treatment with the conventional CD3 binding
molecule (CD19.times.CD3 binding molecule specific for CD19 on B
cells and CD3 on T cells) of those B-NHL patients, who have no
circulating CD19-positive target B cells as described in example
13.
[0506] Continuous infusion was carried out according to the Swivel
method as follows: The monkeys are catheterized via the vena
femoralis into the vena cava caudalis using a vein catheter. The
catheter is tunneled subcutaneously to the dorsal shoulder region
and exteriorized at the caudal scapula. Then a tube is passed
through a jacket and a protection spring. The jacket is fastened
around the animal and the catheter, via the tube, is connected to
an infusion pump.
[0507] Administration solution (1.25 M lysine, 0.1% tween 80, pH 7)
without test material was infused continuously at 48 ml/24 h for 7
days prior to treatment start to allow acclimatization of the
animals to the infusion conditions. Treatment was started by adding
MCSP-G4 VH-VL.times.I2C VH-VL test material to the administration
solution at the amount required for each individual dose level to
be tested (i.e. flow rate of MCSP-G4 VH-VL.times.I2C VH-VL). The
infusion reservoir was changed every day throughout the whole
acclimatization and treatment phase. Treatment duration was 7
days.
[0508] Time courses of absolute T-cell counts in peripheral blood
were determined by four color FACS analysis as follows:
[0509] Collection of Blood Samples and Routine Analysis
[0510] Blood samples (1 ml) were obtained before and 0.75, 2, 6,
12, 24, 30, 48, 72 hours after start of continuous infusion with
MCSP-G4 VH-VL.times.I2C VH-VL as well as after 7 days of treatment
using EDTA-containing Vacutainer.TM. tubes (Becton Dickinson) which
were shipped for analysis at 4.degree. C. In some cases slight
variations of these time points occurred for operational reasons.
FACS analysis of lymphocyte subpopulations was performed within
24-48 h after blood sample collection. Absolute numbers of
leukocyte subpopulations in the blood samples were determined
through differential blood analysis in a routine veterinary
lab.
[0511] Isolation of PBMC from Blood Samples
[0512] PBMC were isolated in analogy to the protocol described in
example 13, above, with adaptations of the used volumes.
[0513] Staining of PBMC with Fluorescence-Labeled Antibodies
Against Cell Surface Molecules
[0514] Monoclonal antibodies reactive with cynomolgus antigens were
obtained from Becton Dickinson (.sup.1Cat. No. 345784, .sup.2Cat.
No. 556647, .sup.3Cat. No. 552851) and Beckman Coulter (.sup.4Cat.
No. IM2470) used according to the manufacturers' recommendations.
5.times.10.sup.5-1.times.10.sup.6 cells were stained with the
following antibody combination: anti-CD14.sup.1
(FITC).times.anti-CD56.sup.2 (PE).times.anti-CD3.sup.3
(PerCP).times.anti-CD19.sup.4 (APC). Additional steps were
performed as described in example 13, above.
[0515] Flowcytometric Detection of Stained Lymphocytes by FACS
[0516] Data collection was performed with a 4 color BD
FACSCalibur.TM. (Becton Dickinson). For each measurement
1.times.10.sup.4 cells of defined lymphocyte subpopulations were
acquired. Statistical analysis was performed with the program
CellQuest Pro.TM. (Becton Dickinson) to obtain lymphocyte
subpopulation percentages and to classify cell surface molecule
expression intensity. Subsequently, percentages of single
lymphocyte subsets related to total lymphocytes (i.e. B plus T plus
NK cells excluding myeloid cells via CD14-staining) as determined
by FACS were correlated with the lymphocyte count from the
differential blood analysis to calculate absolute cell numbers of T
cells (CD3.sup.+, CD56.sup.-, CD14.sup.-).
[0517] T cell redistribution during the starting phase of treatment
with MCSP-G4 VH-VL.times.I2C VH-VL in cynomolgus monkeys at dose
levels of 60, 240 and 1000 .mu.g/m.sup.2/24 h is shown in FIG. 28.
These animals showed no signs at all of any T cell redistribution
during the starting phase of treatment, i.e. T cell counts rather
increased than decreased upon treatment initiation. Given that T
cell redistribution is consistently observed in 100% of all
patients without circulating target cells, upon treatment
initiation with the conventional CD3 binding molecule (e.g.
CD19.times.CD3 construct as described in WO 99/54440) against a
context dependent CD3 epitope, it was demonstrated that
substantially less T cell redistribution in the absence of
circulating target cells upon treatment initiation can be observed
with a CD3 binding molecule of the invention directed and generated
against an epitope of human an non-chimpanzee primate CD3 epsilon
chain as defined by the amino acid sequence of anyone of SEQ ID
NOs: 2, 4, 6, or 8 or a fragment thereof. This is in clear contrast
to CD3-binding molecules directed against a context-dependent CD3
epitope, like the constructs described in WO 99/54440, The binding
molecules against context-independent CD3 epitopes, as (inter alia)
provided in any one of SEQ ID NOs: 2, 4, 6, or 8 (or fragments of
these sequences) provide for this substantially less (detrimental
and non-desired) T cell redistribution. Because T cell
redistribution during the starting phase of treatment with CD3
binding molecules is a major risk factor for CNS adverse events,
the CD3 binding molecules provided herein and capable of
recognizing a context independent CD3 epitope have a substantial
advantage over the CD3 binding molecules known in the art and
directed against context-dependent CD3 epitopes. Indeed none of the
cynomolgus monkeys treated with MCSP-G4 VH-VL.times.I2C VH-VL
showed any signs of CNS symptoms.
[0518] The context-independence of the CD3 epitope is provided in
this invention and corresponds to the first 27 N-terminal amino
acids of CD3 epsilon) or fragments of this 27 amino acid stretch.
This context-independent epitope is taken out of its native
environment within the CD3 complex and fused to heterologous amino
acid sequences without loss of its structural integrity. Anti-CD3
binding molecules as provided herein and generated (and directed)
against a context-independent CD3 epitope provide for a surprising
clinical improvement with regard to T cell redistribution and,
thus, a more favorable safety profile. Without being bound by
theory, since their CD3 epitope is context-independent, forming an
autonomous selfsufficient subdomain without much influence on the
rest of the CD3 complex, the CD3 binding molecules provided herein
induce less allosteric changes in CD3 conformation than the
conventional CD3 binding molecules (like molecules provided in WO
99/54440), which recognize context-dependent CD3 epitopes like
molecules provided in WO 99/54440. As a consequence (again without
being bound by theory), the induction of intracellular NcK2
recruitment by the CD3 binding molecules provided herein is also
reduced resulting in less isoform switch of T cell integrins and
less adhesion of T cells to endothelial cells. It is preferred that
preparations of CD3 binding molecules of the invention (directed
against and generated against a context-independent epitope as
defined herein) essentially consists of monomeric molecules. These
monomeric molecules are even more efficient (than dimeric or
multimeric molecules) in avoiding T cell redistribution and thus
the risk of CNS adverse events during the starting phase of
treatment.
[0519] 16. Generation and Characterization of CD33 and CD3
Cross-Species Specific Bispecific Single Chain Molecules
[0520] 16.1. Generation of CHO Cells Expressing Human CD33
[0521] The coding sequence of human CD33 as published in
GENBANK.RTM. (Accession number NM_001772) was obtained by gene
synthesis according to standard protocols. The gene synthesis
fragment was designed as to contain first a Kozak site for
eukaryotic expression of the construct, followed by a 19 amino acid
immunoglobulin leader peptide, followed in frame by the coding
sequence of the mature human CD33 protein, followed in frame by the
coding sequence of serine glycine dipeptide, a histidine.sub.6-tag
and a stop codon (the cDNA and amino acid sequence of the construct
is listed under SEQ ID Nos 305 and 306). The gene synthesis
fragment was also designed as to introduce restriction sites at the
beginning and at the end of the fragment. The introduced
restriction sites, EcoRI at the 5' end and SalI at the 3' end, were
utilised in the following cloning procedures. The gene synthesis
fragment was cloned via EcoRI and SalI into a plasmid designated
pEF-DHFR (pEF-DHFR is described in Raum et al. Cancer Immunol
Immunother 50 (2001) 141-150) following standard protocols. The
aforementioned procedures were carried out according to standard
protocols (Sambrook, Molecular Cloning; A Laboratory Manual, 3rd
edition, Cold Spring Harbour Laboratory Press, Cold Spring Harbour,
N.Y. (2001)). A clone with sequence-verified nucleotide sequence
was transfected into DHFR deficient CHO cells for eukaryotic
expression of the construct. Eukaryotic protein expression in DHFR
deficient CHO cells (ATCC No. CRL 9096) was performed as described
by Kaufmann R. J. (1990) Methods Enzymol. 185, 537-566. Gene
amplification of the construct was induced by increasing
concentrations of methothrexate (MTX) to a final concentration of
up to 20 nM MTX.
[0522] 16.2. Generation of CHO Cells Expressing the Extracellular
Domain of Macaque CD33
[0523] The cDNA sequence of macaque CD33 was obtained by a set of 3
PCRs on cDNA from macaque monkey bone marrow prepared according to
standard protocols. The following reaction conditions: 1 cycle at
94.degree. C. for 3 minutes followed by 35 cycles with 94.degree.
C. for 1 minute, 53.degree. C. for 1 minute and 72.degree. C. for 2
minutes followed by a terminal cycle of 72.degree. C. for 3 minutes
and the following primers were used:
TABLE-US-00008 1. forward primer: (SEQ ID No. 369)
5'-gaggaattcaccatgccgctgctgctactgctgcccctgctgtgggc
aggggccctggctatgg-3' reverse primer: (SEQ ID No. 370)
5'-gatttgtaactgtatttggtacttcc-3' 2. forward primer: (SEQ ID No.
371) 5'-attccgcctccttggggatcc-3' reverse primer: (SEQ ID No. 372)
5'-gcataggagacattgagctggatgg-3' 3. forward primer: (SEQ ID No. 373)
5'-gcaccaacctgacctgtcagg-3' reverse primer: (SEQ ID No. 374)
5'-agtgggtcgactcactgggtcctgacctctgagtattcg-3'
[0524] Those PCRs generate three overlapping fragments, which were
isolated and sequenced according to standard protocols using the
PCR primers, and thereby provided a portion of the cDNA sequence of
macaque CD33 from the second nucleotide of codon +2 to the third
nucleotide of codon +340 of the mature protein. To generate a
construct for expression of macaque CD33 a cDNA fragment was
obtained by gene synthesis according to standard protocols (the
cDNA and amino acid sequence of the construct is listed under SEQ
ID Nos 307 and 308). In this construct the coding sequence of
macaque CD33 from amino acid +3 to +340 of the mature CD33 protein
was fused into the coding sequence of human CD33 replacing the
human coding sequence of the amino acids +3 to +340. The gene
synthesis fragment was also designed as to contain a Kozak site for
eukaryotic expression of the construct and restriction sites at the
beginning and the end of the fragment containing the cDNA coding
for essentially the whole extracellular domain of macaque CD33, the
macaque CD33 transmembrane domain and a macaque-human chimeric
intracellular CD33 domain. The introduced restriction sites XbaI at
the 5' end and SalI at the 3' end, were utilised in the following
cloning procedures. The gene synthesis fragment was then cloned via
XbaI and SalI into a plasmid designated pEF-DHFR (pEF-DHFR is
described in Raum et al. Cancer Immunol Immunother 50 (2001)
141-150). A sequence verified clone of this plasmid was used to
transfect CHO/dhfr- cells as described above.
[0525] 16.3. Generation of CD33 and CD3 Cross-Species Specific
Bispecific Antibody Molecules
[0526] Cloning of Cross-Species Specific Binding Molecules
[0527] Generally, bispecific antibody molecules, each comprising a
domain with a binding specificity cross-species specific for human
and non-chimpanzee primate CD3 epsilon as well as a domain with a
binding specificity cross-species specific for human and
non-chimpanzee primate CD33, were designed as set out in the
following Table 5:
TABLE-US-00009 TABLE 5 Formats of anti-CD3 and anti-CD33
cross-species specific bispecific molecules SEQ ID Formats of
protein constructs (nucl/prot) (N .fwdarw. C) 276/275 AH11HL
.times. H2CHL 258/257 AH3HL .times. H2CHL 270/269 AC8HL .times.
H2CHL 264/263 AF5HL .times. H2CHL 288/287 F2HL .times. H2CHL
300/299 E11HL .times. H2CHL 282/281 B3HL .times. H2CHL 294/293
B10HL .times. H2CHL 278/277 AH11HL .times. F12QHL 260/259 AH3HL
.times. F12QHL 272/271 AC8HL .times. F12QHL 266/265 AF5HL .times.
F12QHL 290/289 F2HL .times. F12QHL 302/301 E11HL .times. F12QHL
284/283 B3HL .times. F12QHL 296/295 B10HL .times. F12QHL 280/279
AH11HL .times. I2CHL 262/261 AH3HL .times. I2CHL 274/273 AC8HL
.times. I2CHL 268/267 AF5HL .times. I2CHL 292/291 F2HL .times.
I2CHL 304/303 E11HL .times. I2CHL 286/285 B3HL .times. I2CHL
298/297 B10HL .times. I2CHL
[0528] The aforementioned constructs containing the variable
light-chain (L) and variable heavy-chain (H) domains cross-species
specific for human and macaque CD33 and the CD3 specific VH and VL
combinations cross-species specific for human and macaque CD3 were
obtained by gene synthesis. The gene synthesis fragments were
designed and eukaryotic protein expression was performed similar as
described in example 9 for the MCSP and CD3 cross-species specific
single chain molecules.
[0529] The same holds true for the expression and purification of
the CD33 and CD3 cross-species specific single chain molecules.
[0530] In the Western Blot a single band was detected at 52 kD
corresponding to the purified bispecific antibody.
[0531] 16.4. Flow Cytometric Binding Analysis of the CD33 and CD3
Cross-Species Specific Bispecific Antibodies
[0532] In order to test the functionality of the cross-species
specific bispecific antibody constructs regarding the capability to
bind to human and macaque CD33 and CD3, respectively, a FACS
analysis was performed similar to the analysis described for the
analysis of the MCSP and CD3 cross-species specific bispecific
antibodies in example 10 using CHO cells expressing the human or
macaque CD33 extracellular domains (see example 16.1 and 16.2).
[0533] The specific binding of human and non-chimpanzee primate CD3
of the CD3 binding molecules of the invention was clearly
detectable as shown in FIGS. 29(1)-29(12). In the FACS analysis all
constructs show binding to CD3 and CD33 as compared to the
respective negative controls. Cross-species specificity of the
bispecific antibodies to human and macaque CD3 and CD33 antigens is
demonstrated.
[0534] 16.5. Bioactivity of CD33 and CD3 Cross-Species Specific
Bispecific Antibodies
[0535] Bioactivity of the generated bispecific antibodies was
analyzed by chromium 51 (.sup.51Cr) release in vitro cytotoxicity
assays using the CD33 positive cell lines described in Examples
16.1 and 16.2. As effector cells stimulated human CD4/CD56 depleted
PBMC or the macaque T cell line 4119LnPx were used as specified in
the respective figures. The cytotoxicity assays were performed
similar to the setting described for the bioactivity analysis of
the MCSP and CD3 cross-species specific bispecific antibodies in
example 11 using CHO cells expressing the human or macaque CD33
extracellular domains (see example 16.1 and 16.2) as target
cells.
[0536] As shown in FIGS. 30(1)-30(8), all of the generated
cross-species specific bispecific constructs demonstrate cytotoxic
activity against human CD33 positive target cells elicited by
stimulated human CD4/CD56 depleted PBMC and against macaque CD33
positive target cells elicited by the macaque T cell line
4119LnPx.
[0537] 17. Purification of Cross-Species Specific Bispecific Single
Chain Molecules by an Affinity Procedure Based on the Context
Independent CD3 Epsilon Epitope Corresponding to the N-Terminal
Amino Acids 1-27
[0538] 17.1 Generation of an Affinity Column Displaying the
Isolated Context Independent Human CD3 Epsilon Epitope
Corresponding to the N-Terminal Amino Acids 1-27
[0539] The plasmid for expression of the construct 1-27 CD3-Fc
consisting of the 1-27 N-terminal amino acids of the human CD3
epsilon chain fused to the hinge and Fc gamma region of human
immunoglobulin IgG1 described above (Example 3; cDNA sequence and
amino acid sequence of the recombinant fusion protein are listed
under SEQ ID NOs 230 and 229) was transfected into DHFR deficient
CHO cells for eukaryotic expression of the construct. Eukaryotic
protein expression in DHFR deficient CHO cells was performed as
described by Kaufmann R. J. (1990) Methods Enzymol. 185, 537-566.
Gene amplification of the construct was induced by increasing
concentrations of methotrexate (MTX) to a final concentration of up
to 20 nM MTX. After two passages of stationary culture the cells
were grown in roller bottles with nucleoside-free HyQ PF CHO liquid
soy medium (with 4.0 mM L-Glutamine with 0.1% Pluronic F--68;
HyClone) for 7 days before harvest. The cells were removed by
centrifugation and the supernatant containing the expressed protein
was stored at -20.degree. C. For the isolation of the fusion
protein a goat anti-human fc affinity column was prepared according
to standard protocols using a commercially available affinity
purified goat anti-human IgG fc fragment specific antibody with
minimal cross-reaction to bovine, horse, and mouse serum proteins
(Jackson ImmunoResearch Europe Ltd.). Using this affinity column
the fusion protein was isolated out of cell culture supernatant on
an Akta Explorer System (GE Amersham) and eluted by citric acid.
The eluate was neutralized and concentrated. After dialysis against
amine free coupling buffer the purified fusion protein was coupled
to an N-Hydroxy-Succinimide NHS activated 1 ml HiTrap column (GE
Amersham).
[0540] After coupling remaining NHS groups were blocked and the
column was washed and stored at 5.degree. C. in storage buffer
containing 0.1% sodium azide.
[0541] 17.2 Purification of Cross-Species Specific Bispecific
Single Chain Molecules Using a Human CD3 Peptide Affinity
Column
[0542] 200 ml cell culture supernatant of cells expressing
cross-species specific bispecific single chain molecules were 0.2
.mu.m sterile filtered and applied to the CD3 peptide affinity
column using an Akta Explorer system (GE Amersham).
[0543] The column was then washed with phosphate buffered saline
PBS pH 7.4 to wash out unbound sample. Elution was done with an
acidic buffer pH 3.0 containing 20 mM Citric acid and 1 M sodium
chloride. Eluted protein was neutralized immediately by 1 M
Trishydroxymethylamine TRIS pH 8.3 contained in the collection
tubes of the fraction collector.
[0544] Protein analysis was done by SDS PAGE and Western Blot.
[0545] For SDS PAGE BisTris Gels 4-12% are used (Invitrogen). The
running buffer was 1.times.MES-SDS-Puffer (Invitrogen). As protein
standard 15 .mu.l prestained Sharp Protein Standard (Invitrogen)
was applied. Electrophoresis was performed for 60 minutes at 200
volts 120 mA max. Gels were washed in demineralised water and
stained with Coomassie for one hour. Gels are destained in
demineralised water for 3 hours. Pictures are taken with a Syngene
Gel documentation system.
[0546] For Western Blot a double of the SDS PAGE gel was generated
and proteins were electroblotted onto a nitrocellulose membrane.
The membrane was blocked with 2% bovine serum albumin in PBS and
incubated with a biotinylated murine Penta His antibody (Qiagen).
As secondary reagent a streptavidin alkaline phosphatase conjugate
(DAKO) was used. Blots were developed with BCIP/NBT substrate
solution (Pierce).
[0547] As demonstrated in FIGS. 31, 32 and 33 the use of a human
CD3 peptide affinity column as described above allows the highly
efficient purification of the bispecific single chain molecules
from cell culture supernatant. The cross-species specific anti-CD3
single chain antibodies contained in the bispecific single chain
molecules therefore enable via their specific binding properties an
efficient generic one-step method of purification for the
cross-species specific bispecific single chain molecules, without
the need of any tags solely attached for purification purposes.
[0548] 18. Generic Pharmacokinetic Assay for Cross-Species Specific
Bispecific Single Chain Molecules
[0549] 18.1 Production of 1-27 CD3-Fc for Use in the
Pharmacokinetic Assay
[0550] The coding sequence of the 1-27 N-terminal amino acids of
the human CD3 epsilon chain fused to the hinge and Fc gamma region
of human immunoglobulin IgG1 was obtained by gene synthesis
according to standard protocols (cDNA sequence and amino acid
sequence of the recombinant fusion protein are listed under SEQ ID
NOs 309 and 310). The gene synthesis fragment was designed as to
contain first a Kozak site for eukaryotic expression of the
construct, followed by a 19 amino acid immunoglobulin leader
peptide, followed in frame by the coding sequence of the first 27
amino acids of the extracellular portion of the mature human CD3
epsilon chain, followed in frame by the coding sequence of the
hinge region and Fc gamma portion of human IgG1 and a stop codon.
The gene synthesis fragment was also designed and cloned as
described in example 3.1, supra. A clone with sequence-verified
nucleotide sequence was transfected into DHFR deficient CHO cells
for eukaryotic expression of the construct. Eukaryotic protein
expression in DHFR deficient CHO cells was performed as described
in example 9, supra. For the isolation of the fusion protein a goat
anti-human fc affinity column was prepared according to standard
protocols using a commercially available affinity purified goat
anti-human IgG fc fragment specific antibody with minimal
cross-reaction to bovine, horse, and mouse serum proteins (Jackson
ImmunoResearch Europe Ltd.). Using this affinity column the fusion
protein was isolated out of cell culture supernatant on an Akta
Explorer System (GE Amersham) and eluted by citric acid. The eluate
was neutralized and concentrated.
[0551] 18.2 Pharmacokinetic Assay for Cross-Species Specific
Bispecific Single Chain Molecules
[0552] The assay is based on the ECL-ELISA technology using
ruthenium labelled detection on carbon plates measured on a Sektor
Imager device (MSD). In a first step, carbon plates (MSD High Bind
Plate 96 well Cat: L15.times.B-3) were coated with 5 .mu.l/well at
50 ng/ml of the purified 1-27 CD3-Fc described in Example 18.1. The
plate was then dried overnight at 25.degree. C. Subsequently plates
were blocked with 5% BSA (Paesel&Lorei #100568) in PBS at 150
.mu.l/well for 1 h at 25.degree. C. in an incubator while shaking
(700 rpm). In the next step plates were washed three times with
0.05% Tween in PBS. A standard curve in 50% macaque serum in PBS
was generated by serial 1:4 dilution starting at 100 ng/ml of the
respective cross-species specific bispecific single chain molecule
to be detected in the assay. Quality control (QC) samples were
prepared in 50% macaque serum in PBS ranging from 1 ng/ml to 50
ng/ml of the respective cross-species specific bispecific single
chain molecule dependent on the expected sample serum
concentrations. Standard, QC or unknown samples were transferred to
the carbon plates at 10 .mu.l/well and incubated for 90 min at
25.degree. C. in the incubator while shaking (700 rpm).
Subsequently plates were washed three times with 0.05% Tween in
PBS. For detection 25 .mu.l/well of penta-His-Biotin antibody
(Qiagen, 200 .mu.g/ml in 0.05% Tween in PBS) was added and
incubated for 1 h at 25.degree. C. in an incubator while shaking
(700 rpm). In a second detection step 25 .mu.l/well
Streptavidin-SulfoTag solution (MSD; Cat: R32AD-1; Lot: W0010903)
was added and incubated for 1 h at 25.degree. C. in an incubator
while shaking (700 rpm). Subsequently plates were washed three
times with 0.05% Tween in PBS. Finally 150 .mu.l/well MSD Reading
Buffer (MSD, Cat: R9ZC-1) was added and plates were read in the
Sektor Imager device.
[0553] FIGS. 34 and 35 demonstrate the feasibility of detection of
cross-species specific bispecific single chain molecules in serum
samples of macaque monkeys for cross-species specific bispecific
single chain molecules. The cross-species specific anti-CD3 single
chain antibodies contained in the bispecific single chain molecules
enable therefore via their specific binding properties a highly
sensitive generic assay for detection of the cross-species specific
bispecific single chain molecules. The assay set out above can be
used in the context of formal toxicological studies that are needed
for drug development and can be easily adapted for measurement of
patient samples in connection with the clinical application of
cross-species specific bispecific single chain molecules.
[0554] 19. Generation of Recombinant Transmembrane Fusion Proteins
of the N-Terminal Amino Acids 1-27 of CD3 Epsilon from Different
Non-Chimpanzee Primates Fused to EpCAM from Cynomolgus Monkey (1-27
CD3-EpCAM).
[0555] 19.1 Cloning and Expression of 1-27 CD3-EpCAM
[0556] CD3 epsilon was isolated from different non-chimpanzee
primates (marmoset, tamarin, squirrel monkey) and swine. The coding
sequences of the 1-27 N-terminal amino acids of CD3 epsilon chain
of the mature human, common marmoset (Callithrix jacchus),
cottontop tamarin (Saguinus oedipus), common squirrel monkey
(Saimiri sciureus) and domestic swine (Sus scrofa; used as negative
control) fused to the N-terminus of Flag tagged cynomolgus EpCAM
were obtained by gene synthesis according to standard protocols
(cDNA sequence and amino acid sequence of the recombinant fusion
proteins are listed under SEQ ID NOs 231 to 240). The gene
synthesis fragments were designed as to contain first a BsrGI site
to allow for fusion in correct reading frame with the coding
sequence of a 19 amino acid immunoglobulin leader peptide already
present in the target expression vector, which was followed in
frame by the coding sequence of the N-terminal 1-27 amino acids of
the extracellular portion of the mature CD3 epsilon chains, which
was followed in frame by the coding sequence of a Flag tag and
followed in frame by the coding sequence of the mature cynomolgus
EpCAM transmembrane protein. The gene synthesis fragments were also
designed to introduce a restriction site at the end of the cDNA
coding for the fusion protein. The introduced restriction sites
BsrGI at the 5' end and SalI at the 3' end, were utilized in the
following cloning procedures. The gene synthesis fragments were
then cloned via BsrGI and SalI into a derivative of the plasmid
designated pEF-DHFR (pEF-DHFR is described in Raum et al. Cancer
Immunol Immunother 50 (2001) 141-150), which already contains the
coding sequence of the 19 amino acid immunoglobulin leader peptide
following standard protocols. Sequence verified plasmids were used
to transfect DHFR deficient CHO cells for eukaryotic expression of
the construct. Eukaryotic protein expression in DHFR deficient CHO
cells was performed as described by Kaufmann R. J. (1990) Methods
Enzymol. 185, 537-566. Gene amplification of the construct was
induced by increasing concentrations of methotrexate (MTX) to a
final concentration of up to 20 nM MTX.
[0557] Transfectants were tested for cell surface expression of the
recombinant transmembrane protein via an FACS assay according to
standard protocols. For that purpose a number of 2.5.times.10.sup.5
cells were incubated with 50 .mu.l of the anti-Flag M2 antibody
(Sigma-Aldrich Chemie GmbH, Taufkirchen, Germany) at 5 .mu.g/ml in
PBS with 2% FCS. Bound antibody was detected with an
R-Phycoerythrin-conjugated affinity purified F(ab')2 fragment, goat
anti-mouse IgG, Fc-gamma fragment specific 1:100 in PBS with 2% FCS
(Jackson ImmunoResearch Europe Ltd., Newmarket, Suffolk, UK). Flow
cytometry was performed on a FACS-Calibur apparatus, the CellQuest
software was used to acquire and analyze the data (Becton Dickinson
biosciences, Heidelberg). FACS staining and measuring of the
fluorescence intensity were performed as described in Current
Protocols in Immunology (Coligan, Kruisbeek, Margulies, Shevach and
Strober, Wiley-Interscience, 2002).
[0558] Expression of the Flag tagged recombinant transmembrane
fusion proteins consisting of cynomolgus EpCAM and the 1-27
N-terminal amino acids of the human, marmoset, tamarin, squirrel
monkey and swine CD3 epsilon chain respectively on transfected
cells is clearly detectable (FIG. 36).
[0559] 19.2 Cloning and Expression of the Cross-Species Specific
Anti-CD3 Single Chain Antibody I2C HL in Form of an IgG1
Antibody
[0560] In order to provide improved means of detection of binding
of the cross-species specific single chain anti-CD3 antibody the
I2C VHVL specificity is converted into an IgG1 antibody with murine
IgG1 and murine kappa constant regions. cDNA sequences coding for
the heavy chain of the IgG antibody were obtained by gene synthesis
according to standard protocols. The gene synthesis fragments were
designed as to contain first a Kozak site to allow for eukaryotic
expression of the construct, which is followed by an 19 amino acid
immunoglobulin leader peptide, which is followed in frame by the
coding sequence of the heavy chain variable region or light chain
variable region, followed in frame by the coding sequence of the
heavy chain constant region of murine IgG1 as published in
GENBANK.RTM. (Accession number AB097849) or the coding sequence of
the murine kappa light chain constant region as published in
GENBANK.RTM. (Accession number D14630), respectively.
[0561] Restriction sites were introduced at the beginning and the
end of the cDNA coding for the fusion protein. Restriction sites
EcoRI at the 5' end and SalI at the 3' end were used for the
following cloning procedures. The gene synthesis fragments were
cloned via EcoRI and SalI into a plasmid designated pEF-DHFR
(pEF-DHFR is described in Raum et al. Cancer Immunol Immunother 50
(2001) 141-150) for the heavy chain construct and pEFADA (pEFADA is
described in Raum et al. loc cit.) for the light chain construct
according to standard protocols. Sequence verified plasmids were
used for co-transfection of respective light and heavy chain
constructs into DHFR deficient CHO cells for eukaryotic expression
of the construct. Eukaryotic protein expression in DHFR deficient
CHO cells was performed as described by Kaufmann R. J. (1990)
Methods Enzymol. 185, 537-566. Gene amplification of the constructs
was induced by increasing concentrations of methotrexate (MTX) to a
final concentration of up to 20 nM MTX and deoxycoformycin (dCF) to
a final concentration of up to 300 nM dCF. After two passages of
stationary culture cell culture supernatant was collected and used
in the subsequent experiment.
[0562] 19.3 Binding of the Cross-Species Specific Anti-CD3 Single
Chain Antibody I2C HL in Form of an IgG1 Antibody to 1-27
CD3-EpCAM
[0563] Binding of the generated I2C IgG1 construct to the 1-27
N-terminal amino acids of the human, marmoset, tamarin and squirrel
monkey CD3 epsilon chains respectively fused to cynomolgus Ep-CAM
as described in Example 19.1 was tested in a FACS assay according
to standard protocols. For that purpose a number of
2.5.times.10.sup.5 cells were incubated with 50 .mu.l of cell
culture supernatant containing the I2C IgG1 construct as described
in Example 19.2. The binding of the antibody was detected with an
R-Phycoerythrin-conjugated affinity purified F(ab')2 fragment, goat
anti-mouse IgG, Fc-gamma fragment specific, diluted 1:100 in PBS
with 2% FCS (Jackson ImmunoResearch Europe Ltd., Newmarket,
Suffolk, UK). Flow cytometry was performed on a FACS-Calibur
apparatus, the CellQuest software was used to acquire and analyze
the data (Becton Dickinson biosciences, Heidelberg). FACS staining
and measuring of the fluorescence intensity were performed as
described in Current Protocols in Immunology (Coligan, Kruisbeek,
Margulies, Shevach and Strober, Wiley-Interscience, 2002).
[0564] As shown in FIG. 37 binding of the I2C IgG1 construct to the
transfectants expressing the recombinant transmembrane fusion
proteins consisting of the 1-27 N-terminal amino acids of CD3
epsilon of human, marmoset, tamarin or squirrel monkey fused to
cynomolgus EpCAM as compared to the negative control consisting of
the 1-27 N-terminal amino acids of CD3 epsilon of swine fused to
cynomolgus EpCAM was observed. Thus multi-primate cross-species
specificity of I2C was demonstrated. Signals obtained with the anti
Flag M2 antibody and the I2C IgG1 construct were comparable,
indicating a strong binding activity of the cross-species specific
specificity I2C to the N-terminal amino acids 1-27 of CD3
epsilon.
[0565] 20. Binding of the Cross-Species Specific Anti-CD3 Binding
Molecule I2C to the Human CD3 Epsilon Chain with and without
N-Terminal His6 Tag
[0566] A chimeric IgG1 antibody with the binding specificity I2C as
described in Example 19.2 specific for CD3 epsilon was tested for
binding to human CD3 epsilon with and without N-terminal His6 tag.
Binding of the antibody to the EL4 cell lines transfected with
His6-human CD3 epsilon as described in Example 6.1 and wild-type
human CD3 epsilon as described in Example 5.1 respectively was
tested by a FACS assay according to standard protocols.
2.5.times.10.sup.5 cells of the transfectants were incubated with
50 .mu.l of cell culture supernatant containing the I2C IgG1
construct or 50 .mu.l of the respective control antibodies at 5
.mu.g/ml in PBS with 2% FCS. As negative control an appropriate
isotype control and as positive control for expression of the
constructs the CD3 specific antibody UCHT-1 were used respectively.
Detection of the His6 tag was performed with the penta His antibody
(Qiagen). The binding of the antibodies was detected with a
R-Phycoerythrin-conjugated affinity purified F(ab')2 fragment, goat
anti-mouse IgG, Fc-gamma fragment specific, diluted 1:100 in PBS
with 2% FCS (Jackson ImmunoResearch Europe Ltd., Newmarket,
Suffolk, UK). Flow cytometry was performed on a FACS-Calibur
apparatus, the CellQuest software was used to acquire and analyze
the data (Becton Dickinson biosciences, Heidelberg). FACS staining
and measuring of the fluorescence intensity were performed as
described in Current Protocols in Immunology (Coligan, Kruisbeek,
Margulies, Shevach and Strober, Wiley-Interscience, 2002).
[0567] Comparable binding of the anti-human CD3 antibody UCHT-1 to
both transfectants demonstrates approximately equal levels of
expression of the constructs. The binding of the penta His antibody
confirmed the presence of the His6 tag on the His6-human CD3
construct but not on the wild-type construct.
[0568] Compared to the EL4 cell line transfected with wild-type
human CD3 epsilon a clear loss of binding of the I2C IgG1 construct
to human-CD3 epsilon with an N-terminal His6 tag was detected.
These results show that a free N-terminus of CD3 epsilon is
essential for binding of the cross-species specific anti-CD3
binding specificity I2C to the human CD3 epsilon chain (FIGS.
28(1)-28(2)).
[0569] 21. Generation of CD33 and CD3 Cross-Species Specific
Bispecific Single Chain Molecules
[0570] 21.1 Generation of CD33 and CD3 Cross-Species Specific
Bispecific Single Chain Molecules
[0571] Generally, bispecific single chain antibody molecules, each
comprising a domain with a binding specificity cross-species
specific for human and macaque CD3epsilon as well as a domain with
a binding specificity cross-species specific for human and macaque
CD33, were designed as set out in the following Table 6:
TABLE-US-00010 TABLE 6 Formats of anti-CD3 and anti-CD33
cross-species specific bispecific single chain antibody molecules
SEQ ID Formats of protein constructs (nucl/prot) (N .fwdarw. C)
316/315 I2CHL .times. AF5HL 314/313 F12QHL .times. AF5HL 312/311
H2CHL .times. AF5HL
[0572] The aforementioned constructs containing the variable
light-chain (L) and variable heavy-chain (H) domains cross-species
specific for human and macaque CD33 and the CD3 specific VH and VL
combinations cross-species specific for human and macaque CD3 were
obtained by gene synthesis. The gene synthesis fragments were
designed in analogy to the procedure described in example 9 for the
MCSP and CD3 cross-species specific single chain molecules. A clone
with sequence-verified nucleotide sequence was transfected into
DHFR deficient CHO cells for eukaryotic expression of the
construct. Eukaryotic protein expression in DHFR deficient CHO
cells was performed as also described in example 9 for the MCSP and
CD3 cross-species specific single chain molecules and used in the
subsequent experiments.
[0573] 21.2 Flow Cytometric Binding Analysis of the CD33 and CD3
Cross-Species Specific Bispecific Antibodies
[0574] In order to test the functionality of the cross-species
specific bispecific antibody constructs regarding the capability to
bind to human and macaque CD33 and CD3, respectively, a FACS
analysis is performed similar to the analysis described for the
analysis of the MCSP and CD3 cross-species specific bispecific
antibodies in example 10 using CHO cells expressing the human or
macyque CD33 extracellular domains (see examples 16.1 and
16.2).
[0575] The bispecific binding of the single chain molecules listed
above, which were cross-species specific for CD33 and cross-species
specific for human and non-chimpanzee primate CD3 was clearly
detectable as shown in FIGS. 41A-41B. In the FACS analysis all
constructs showed binding to CD3 and CD33 as compared to the
respective negative controls. Cross-species specificity of the
bispecific antibodies to human and macaque CD3 and CD33 antigens
was demonstrated.
[0576] 21.3. Bioactivity of CD33 and CD3 Cross-Species Specific
Bispecific Single Chain Antibodies
[0577] Bioactivity of the generated bispecific single chain
antibodies was analyzed by chromium 51 (.sup.51Cr) release in vitro
cytotoxicity assays using the CD33 positive cell lines described in
Examples 16.1 and 16.2. As effector cells stimulated human CD4/CD56
depleted PBMC or the macaque T cell line 4119LnPx were used as
specified in the respective figures. The cytotoxicity assays were
performed similar to the procedure described for the bioactivity
analysis of the MCSP and CD3 cross-species specific bispecific
antibodies in example 11 using CHO cells expressing the human or
macaque CD33 extracellular domains (see example 16.1 and 16.2) as
target cells.
[0578] As shown in FIG. 42, all of the generated cross-species
specific bispecific single chain antibody constructs demonstrate
cytotoxic activity against human CD33 positive target cells
elicited by stimulated human CD4/CD56 depleted PBMC and against
macaque CD33 positive target cells elicited by the macaque T cell
line 4119LnPx.
[0579] 22. Redistribution of Circulating Chimpanzee T Cells Upon
Exposure to a Conventional Bispecific CD3 Binding Molecule Directed
at a Target Molecule which is Absent from Circulating Blood
Cells
[0580] A single male chimpanzee was subjected to dose escalation
with intravenous single-chain EpCAM/CD3-bispecific antibody
construct (Schlereth (2005) Cancer Res 65: 2882). Like in the
conventional single-chain CD19/CD3-bispecific antibody construct
(Loffler (2000, Blood, Volume 95, Number 6) or WO 99/54440), the
CD3 arm of said EpCAM/CD3-construct is also directed against a
conventional context dependent epitope of human and chimpanzee CD3.
At day 0, the animal received 50 ml PBS/5% HSA without test
material, followed by 50 ml PBS/5% HSA plus single-chain
EpCAM/CD3-bispecific antibody construct at 1.6, 2.0, 3.0 and 4.5
.mu.g/kg on days 7, 14, 21 and 28, respectively. The infusion
period was 2 hours per administration. For each weekly infusion the
chimpanzee was sedated with 2-3 mg/kg Telazol intramuscularly,
intubated and placed on isoflurane/O.sub.2 anesthesia with stable
mean blood pressures. A second intravenous catheter was placed in
an opposite limb to collect (heparinized) whole blood samples at
the time points indicated in FIG. 43 for FACS analysis of
circulating blood cells. After standard erythrocyte lysis, T cells
were stained with a FITC-labeled antibody reacting with chimpanzee
CD2 (Becton Dickinson) and the percentage of T cells per total
lymphocytes determined by flow cytometry. As shown in FIG. 43,
every administration of single-chain EpCAM/CD3-bispecific antibody
construct induced a rapid drop of circulating T cells as observed
with single-chain CD19/CD3-bispecific antibody construct in B-NHL
patients, who had essentially no circulating target B (lymphoma)
cells. As there are no EpCAM-positive target cells in the
circulating blood of humans and chimpanzees, the drop of
circulating T cells upon exposure to the single-chain
EpCAM/CD3-bispecific antibody construct can be attributed solely to
a signal, which the T cells receive through pure interaction of the
CD3 arm of the construct with a conventional context dependent CD3
epitope in the absence of any target cell mediated crosslinking.
Like the redistribution of T cells induced through their exposure
to single-chain CD19/CD3-bispecific antibody construct in B-NHL
patients, who had essentially no circulating target B (lymphoma)
cells, the T cell redistribution in the chimpanzee upon exposure to
the single-chain EpCAM/CD3-bispecific antibody construct can be
explained by a conformational change of CD3 following the binding
event to a context dependent CD3 epitope further resulting in the
transient increase of T cell adhesiveness to blood vessel
endothelium (see Example 13). This finding confirms, that
conventional CD3 binding molecules directed to context dependent
CD3 epitopes--solely through this interaction--can lead to a
redistribution pattern of peripheral blood T cells, which is
associated with the risk of CNS adverse events in humans as
describe in Example 13.
[0581] 23. Specific Binding of scFv Clones to the N-Terminus of
Human CD3 Epsilon
[0582] 23.1 Bacterial Expression of scFv Constructs in E. coli XL1
Blue
[0583] As previously mentioned, E. coli XL1 Blue transformed with
pComb3H5Bhis/Flag containing a VL- and VH-segment produce soluble
scFv in sufficient amounts after excision of the gene III fragment
and induction with 1 mM IPTG. The scFv-chain is exported into the
periplasma where it folds into a functional conformation.
[0584] The following scFv clones were chosen for this
experiment:
[0585] i) ScFvs 4-10, 3-106, 3-114, 3-148, 4-48, 3-190 and 3-271 as
described in WO 2004/106380.
[0586] ii) ScFvs from the human anti-CD3epsilon binding clones H2C,
F12Q and I2C as described herein.
[0587] For periplasmic preparations, bacterial cells transformed
with the respective scFv containing plasmids allowing for
periplasmic expression were grown in SB-medium supplemented with 20
mM MgCl.sub.2 and carbenicillin 50 .mu.g/ml and redissolved in PBS
after harvesting. By four rounds of freezing at -70.degree. C. and
thawing at 37.degree. C., the outer membrane of the bacteria was
destroyed by osmotic shock and the soluble periplasmic proteins
including the scFvs were released into the supernatant. After
elimination of intact cells and cell-debris by centrifugation, the
supernatant containing the human anti-human CD3-scFvs was collected
and used for further examination. These crude supernatants
containing scFv will be further termed periplasmic preparations
(PPP).
[0588] 23.2 Binding of scFvs to Human CD3 Epsilon (aa 1-27)-Fc
Fusion Protein
[0589] ELISA experiments were carried out by coating the human CD3
epsilon (aa 1-27)-Fc fusion protein to the wells of 96 well plastic
plates (Nunc, maxisorb) typically at 4.degree. C. over night. The
antigen coating solution was then removed, wells washed once with
PBS/0.05% Tween 20 and subsequently blocked with PBS/3% BSA for at
least one hour. After removal of the blocking solution, PPPs and
control solutions were added to the wells and incubated for
typically one hour at room temperature. The wells were then washed
three times with PBS/0.05% Tween 20. Detection of scFvs bound to
immobilized antigen was carried out using a Biotin-labeled anti
FLAG-tag antibody (M2 anti Flag-Bio, Sigma, typically at a final
concentration of 1 .mu.g/ml PBS) and detected with a
peroxidase-labeled Streptavidine (Dianova, 1 .mu.g/ml PBS). The
signal was developed by adding ABTS substrate solution and measured
at a wavelength of 405 nm. Unspecific binding of the test-samples
to the blocking agent and/or the human IgG1 portion of the human
CD3 epsilon (aa 1-27)-Fc fusion protein was examined by carrying
out the identical assay with the identical reagents and identical
timing on ELISA plates which were coated with human IgG1 (Sigma).
PBS was used as a negative control.
[0590] As shown in FIG. 44, scFvs H2C, F12Q and I2C show strong
binding signals on human CD3 epsilon (aa 1-27)-Fc fusion protein.
The human scFvs 3-106, 3-114, 3-148, 3-190, 3-271, 4-10 and 4-48
(as described in WO 2004/106380) do not show any significant
binding above negative control level.
[0591] To exclude the possibility that the positive binding of
scFvs H2C, F12Q and I2C to wells coated with human CD3 epsilon (aa
1-27)-Fc fusion protein might be due to binding to BSA (used as a
blocking agent) and/or the human IgG1 Fc-gamma-portion of the human
CD3 epsilon (aa 1-27)-Fc fusion protein, a second ELISA experiment
was performed in parallel. In this second ELISA experiment, all
parameters were identical to those in the first ELISA experiment,
except that in the second ELISA experiment human IgG1 (Sigma) was
coated instead of human CD3 epsilon (aa 1-27)-Fc fusion protein. As
shown in FIG. 45, none of the scFvs tested showed any significant
binding to BSA and/or human IgG1 above background level.
[0592] Taken together, these results allow the conclusion that
conventional CD3 binding molecules recognizing a context-dependent
epitope of CD3 epsilon (e.g. as disclosed in WO 2004/106380) do not
bind specifically to the human CD3 epsilon (aa 1-27)-region,
whereas the scFvs H2C, F12Q and I2C binding a context-independent
epitope of CD3 epsilon clearly show specific binding to the
N-terminal 27 amino acids of human CD3 epsilon.
[0593] 24. Generation and Characterization of PSMA and CD3
Cross-Species Specific Bispecific Single Chain Antibody
Molecules
[0594] 24.1 Cloning and Expression of Human PSMA Antigen on CHO
Cells
[0595] The sequence of the human PSMA antigen (`AY101595`, Homo
sapiens prostate-specific membrane antigen mRNA, complete cds,
National Center for Biotechnology Information,
http_colon-slash-slash_www.ncbi.nlm.nih.gov/entrez) was used to
obtain a synthetic molecule by gene synthesis according to standard
protocols. The gene synthesis fragment was also designed as to
contain a Kozak site for eukaryotic expression of the construct and
restriction sites at the beginning and the end of the DNA. The
introduced restriction sites XbaI at the 5' end and SalI at the 3'
end were utilised during the cloning step into the expression
plasmid designated pEFDHFR as described in Mack et al. (Mack M et
al., Proc Natl Acad Sci USA 1995; 92:7021-5. and Raum et al. Cancer
Immunol Immunother (2001) 50(3)). After sequence verification the
plasmid was used to transfect CHO/dhfr- cells as follows. A
sequence verified plasmid was used to transfect CHO/dhfr- cells
(ATCC No. CRL 9096; cultivated in RPMI 1640 with stabilized
glutamine obtained from Biochrom AG Berlin, Germany, supplemented
with 10% FCS, 1% penicillin/streptomycin all obtained from Biochrom
AG Berlin, Germany and nucleosides from a stock solution of cell
culture grade reagents obtained from Sigma-Aldrich Chemie GmbH,
Taufkirchen, Germany, to a final concentration of 10 .mu.g/ml
Adenosine, 10 .mu.g/ml Deoxyadenosine and 10 .mu.g/ml Thymidine, in
an incubator at 37.degree. C., 95% humidity and 7% CO.sub.2).
Transfection was performed using the PolyFect Transfection Reagent
(Qiagen GmbH, Hilden, Germany) and 5 .mu.g of plasmid DNA according
to the manufacturer's protocol. After a cultivation of 24 hours
cells were washed once with PBS and again cultivated in the
aforementioned cell culture medium except that the medium was not
supplemented with nucleosides and dialysed FCS (obtained from
Biochrom AG Berlin, Germany) was used. Thus the cell culture medium
did not contain nucleosides and thereby selection was applied on
the transfected cells. Approximately 14 days after transfection the
outgrowth of resistant cells was observed. After an additional 7 to
14 days the transfectants were tested positive for expression of
the construct via FACS. Eukaryotic protein expression in DHFR
deficient CHO cells is performed as described by Kaufmann R. J.
(1990) Methods Enzymol. 185, 537-566. Gene amplification of the
construct is induced by increasing concentrations of methothrexate
(MTX) to a final concentration of up to 20 nM MTX
[0596] 24.2 Cloning and Expression of Macaque PSMA Antigen on CHO
Cells
[0597] The cDNA sequence of macaque PSMA (cynomolgus) was obtained
by a set of five PCRs on cDNA from macaque monkey prostate prepared
according to standard protocols. The following reaction conditions:
1 cycle at 94.degree. C. for 2 minutes followed by 40 cycles with
94.degree. C. for 1 minute, 52.degree. C. for 1 minute and
72.degree. C. for 1.5 minutes followed by a terminal cycle of
72.degree. C. for 3 minutes and the following primers were
used:
TABLE-US-00011 4. forward primer: (SEQ ID NO. 375)
5'-cactgtggcccaggttcgagg-3' reverse primer: (SEQ ID NO. 376)
5'-gacataccacacaaattcaatacgg-3' 5. forward primer: (SEQ ID NO. 377)
5'-gctctgctcgcgccgagatgtgg-3' reverse primer: (SEQ ID NO. 378)
5'-acgctggacaccacctccagg-3' 6. forward primer: (SEQ ID NO. 379)
5'-ggttctactgagtgggcagagg-3' reverse primer: (SEQ ID NO. 380)
5'-acttgttgtggctgcttggagc-3' 7. forward primer: (SEQ ID NO. 381)
5'-gggtgaagtcctatccagatgg-3' reverse primer: (SEQ ID NO. 382)
5'-gtgctctgcctgaagcaattcc-3' 8. forward primer: (SEQ ID NO. 383)
5'-ctcggcttcctcttcgggtgg-3' reverse primer: (SEQ ID NO. 384)
5'-gcatattcatttgctgggtaacctgg-3'
[0598] These PCRs generated five overlapping fragments, which were
isolated and sequenced according to standard protocols using the
PCR primers, and thereby provided a portion of the cDNA sequence
coding macaque PSMA from codon 3 to the last codon of the mature
protein. To generate a construct for expression of macaque PSMA a
cDNA fragment was obtained by gene synthesis according to standard
protocols (the cDNA and amino acid sequence of the construct is
listed under SEQ ID 385 and 386). In this construct the coding
sequence of macaque PSMA from amino acid 3 to the last amino acid
of the mature PSMA protein followed by a stop codon was fused in
frame to the coding sequence of the first two amino acids of the
human PSMA protein. The gene synthesis fragment was also designed
as to contain a Kozak site for eukaryotic expression of the
construct and restriction sites at the beginning and the end of the
fragment containing the cDNA. The introduced restriction sites,
XbaI at the 5' end and SalI at the 3' end, were utilised in the
following cloning procedures. The gene synthesis fragment was
cloned via XbaI and SalI into a plasmid designated pEF-DHFR
following standard protocols. The aforementioned procedures were
carried out according to standard protocols (Sambrook, Molecular
Cloning; A Laboratory Manual, 3rd edition, Cold Spring Harbour
Laboratory Press, Cold Spring Harbour, N.Y. (2001)). A clone with
sequence-verified nucleotide sequence was transfected into DHFR
deficient CHO cells for eukaryotic expression of the construct.
Eukaryotic protein expression in DHFR deficient CHO cells was
performed as described by Kaufmann R. J. (1990) Methods Enzymol.
185, 537-566. Gene amplification of the construct was induced by
increasing concentrations of methotrexate (MTX) to a final
concentration of up to 20 nM MTX.
[0599] 24.3 Generation of PSMA and CD3 Cross-Species Specific
Bispecific Single Chain Molecules
[0600] Generally, bispecific single chain antibody molecules, each
comprising a domain with a binding specificity for the human and
the macaque CD3 antigen as well as a domain with a binding
specificity for the human and the macaque PSMA antigen, were
designed as set out in the following Table 7:
TABLE-US-00012 TABLE 7 Formats of anti-CD3 and anti-PSMA
cross-species specific bispecific single chain antibody molecules
SEQ ID NO. Formats of protein constructs (nucl/prot) (N .fwdarw. C)
400/399 PSMA-3 HL .times. I2C HL 414/413 PSMA-4 HL .times. I2C HL
428/427 PSMA-6 LH .times. I2C HL 442/441 PSMA-7 LH .times. I2C HL
456/455 PSMA-8 LH .times. I2C HL 470/469 PSMA-9 LH .times. I2C HL
484/483 PSMA-10 LH .times. I2C HL 498/497 PSMA-A LH .times. I2C HL
512/511 PSMA-B LH .times. I2C HL 526/525 PSMA-C LH .times. I2C HL
540/539 PSMA-D LH .times. I2C HL 554/553 PSMA-E LH .times. I2C HL
568/567 PSMA-F LH .times. I2C HL 582/581 PSMA-J LH .times. I2C HL
596/595 PSMA-L LH .times. I2C HL
[0601] The aforementioned constructs containing the variable
light-chain (L) and variable heavy-chain (H) domains cross-species
specific for human and macaque PSMA and the CD3 specific VH and VL
combinations cross-species specific for human and macaque CD3 were
obtained by gene synthesis. The gene synthesis fragments were
designed and eukaryotic protein expression was performed in analogy
to the procedure described in example 9 for the MCSP and CD3
cross-species specific single chain molecules. Alternatively the
constructs can be transfected into DHFR-deficient CHO-cells in a
transient manner according to standard protocols.
[0602] 24.4 Flow Cytometric Binding Analysis of the PSMA and CD3
Cross-Species Specific Bispecific Antibodies
[0603] In order to test the functionality of the cross-species
specific bispecific antibody constructs with regard to binding
capability to human and macaque PSMA and to human and macaque CD3,
a FACS analysis was performed. For this purpose the CHO cells
transfected with human PSMA as described in Example 24.1 and human
CD3 positive T cell leukemia cell line HPB-ALL (DSMZ, Braunschweig,
ACC483) were used to check the binding to human antigens. The
binding reactivity to macaque antigens was tested by using the
generated macaque PSMA transfectant described in Example 24.2 and a
macaque T cell line 4119LnPx (kindly provided by Prof Fickenscher,
Hygiene Institute, Virology, Erlangen-Nuernberg; published in
Knappe A, et al., and Fickenscher H., Blood 2000, 95, 3256-61). The
flow cytrometric analysis was performed in analogy to the procedure
described in example 10.
[0604] The binding ability of all PSMA based bispecific single
chain molecules were clearly detectable as shown in FIGS.
46(1)-46(4). In the FACS analysis, all constructs showed binding to
CD3 and PSMA compared to the negative control using culture medium
and 1. and 2. detection antibody. In summary, the cross-species
specificity of the bispecific antibody to human and macaque CD3 and
to human and macaque PSMA could clearly be demonstrated.
[0605] 24.5 Bioactivity of PSMA and CD3 Cross-Species Specific
Bispecific Single Chain Antibodies
[0606] Bioactivity of the generated bispecific single chain
antibodies was analyzed by chromium 51 release in vitro
cytotoxicity assays using the PSMA positive cell lines described in
example 24.1 and 24.2. As effector cells stimulated human CD8
positive T cells or the macaque T cell line 4119LnPx were used. The
cytotoxicity assays were performed similar to the procedure
described for the bioactivity analysis of the MCSP and CD3
cross-species specific bispecific antibodies in example 11.
[0607] As shown in FIGS. 47(1)A-47(2)B and 48(1)A-48(2)B, all of
the depicted cross-species specific bispecific single chain
antibody constructs revealed cytotoxic activity against human PSMA
positive target cells elicited by human CD8+ cells and against
macaque PSMA positive target cells elicited by the macaque T cell
line 4119LnPx. As a negative control, an irrelevant bispecific
single chain antibody was used.
[0608] 24.6 Generation of PSMA and CD3 Cross-Species Specific
Bispecific Single Chain Molecules
[0609] Bispecific single chain antibody molecules, each comprising
a domain binding to the human and to the macaque CD3 antigen as
well as a domain binding to the human PSMA antigen, were designed
as set out in the following Table 8:
TABLE-US-00013 TABLE 8 Formats of anti-CD3 and anti-PSMA
cross-species specific bispecific single chain antibody molecules
SEQ ID NO. Formats of protein constructs (nucl/prot) (N .fwdarw. C)
610/609 PM99-A8 HL .times. I2C HL 624/623 PM86-A10 HL .times. I2C
HL 638/637 PM86-B4-2 HL .times. I2C HL 652/651 PM98-B4 HL .times.
I2C HL 666/665 PM86-C3 HL .times. I2C HL 680/679 PM86-E12 HL
.times. I2C HL 694/693 PMF1-A10 HL .times. I2C HL 708/707 PM99-F1
HL .times. I2C HL 736/721 PM99-F5 HL .times. I2C HL 735/734 PM86-F6
HL .times. I2C HL 800/799 PM76-A9 HL .times. I2C HL 818/817
PM76-B10 HL .times. I2C HL 864/863 PM29-G1 HL .times. I2C HL
850/849 PM49-B9 HL .times. I2C HL 836/835 PM34-C7 HL .times. I2C HL
786/785 PM84-D7 HL .times. I2C HL 882/881 PM08-B6 HL .times. I2C HL
900/899 PM08-E11 HL .times. I2C HL 936/935 PM95-A8 HL .times. I2C
HL 1018/1017 PM26-C9 HL .times. I2C HL 1032/1031 PM26-H4 HL .times.
I2C HL 918/917 PM95-H6 HL .times. I2C HL 1004/1003 PM07-D3 HL
.times. I2C HL 954/953 PM07-A12 HL .times. I2C HL 972/971 PM07-F8
HL .times. I2C HL 990/989 PM07-E5 HL .times. I2C HL
[0610] The aforementioned constructs each comprising a combination
of a variable light-chain (L) and a variable heavy-chain (H) domain
binding to the human and to the macaque CD3 antigen as well as a
combination of a variable light-chain (L) and a variable
heavy-chain (H) domains binding to the human PSMA antigen were
obtained by gene synthesis. Each combination of a variable
light-chain (L) and a variable heavy-chain (H) domains binding to
the human PSMA antigen was obtained via phage display from a
scFv-library by panning on the PSMA-positive human prostate cancer
cell line LNCaP (ATCC No. CRL-1740) followed by FACS-based
screening for positive clones using the same cell line. The gene
synthesis fragments of the above listed bispecific single chain
antibody molecules were designed and eukaryotic protein expression
was performed in analogy to the procedure described in example
24.3, supra, respectively for the MCSP and CD3 cross-species
specific single chain molecules in example 9. The same holds true
for the expression and purification of the PSMA and CD3 bispecific
single chain antibody molecules.
[0611] 24.7 Flow Cytometric Binding Analysis of PSMA and CD3
Cross-Species Specific Bispecific Antibodies
[0612] In order to test the functionality of cross-species specific
bispecific antibody constructs regarding the capability to bind to
PSMA and CD3 a FACS analysis was performed. For this purpose
PSMA-positive cells were used to test the binding to human
antigens. The binding reactivity to macaque CD3 was tested by using
the macaque T cell line 4119LnPx (kindly provided by Prof
Fickenscher, Hygiene Institute, Virology, Erlangen-Nuernberg;
published in Knappe A, et al., and Fickenscher H., Blood 2000, 95,
3256-61). The flow cytrometric analysis was performed in analogy to
the procedure described in example 10.
[0613] The bispecific binding of the generated single chain
molecules shown in FIGS. 49A-49E and FIGS. 51A-51C, to human PSMA
and to human and non-chimpanzee primate CD3 was clearly detectable.
In the FACS analysis all shown constructs demonstrated binding to
CD3 and PSMA compared to the negative control.
[0614] 24.8 Bioactivity of PSMA and CD3 Cross-Species Specific
Bispecific Single Chain Antibodies
[0615] Bioactivity of generated bispecific single chain antibodies
was analyzed by chromium 51 (.sup.51Cr) release in vitro
cytotoxicity assays using PSMA positive cell lines. As effector
cells stimulated human CD4/CD56 depleted PBMC or the macaque T cell
line 4119LnPx were used. The cytotoxicity assays were performed
similar to the procedure described for the bioactivity analysis of
the MCSP and CD3 cross-species specific bispecific antibodies in
example 11.
[0616] The generated cross-species specific bispecific single chain
antibody constructs shown in FIGS. 50(1)-50(3)B and 52A-52B
demonstrated cytotoxic activity against PSMA positive target
cells.
[0617] 24.9. Generation of Additional PSMA and CD3 Cross-Species
Specific Bispecific Single Chain Molecules
[0618] The human antibody germline VH sequence VH3 3-11
(_http_colon-slash-slash vbase.mrc-cpe.cam.ac.uk) is chosen as
framework context for CDRH1 (SEQ ID NO. 394, CDRH2 (SEQ ID NO. 395)
and CDRH3 (SEQ ID NO. 396). Likewise the human antibody germline VH
sequence VH1 1-02 (http colon-slash-slash vbase.mrc-cpe.cam.ac.uk)
is chosen as framework context for CDRH1 (SEQ ID NO. 408), CDRH2
(SEQ ID NO. 409) and CDRH3 (SEQ ID NO. 410) as well as the human
antibody germline VH sequence VH1 1-03 (http colon-slash-slash
vbase.mrc-cpe.cam.ac.uk) as framework context for CDRH1 (SEQ ID NO.
445), CDRH2 (SEQ ID NO. 446) and CDRH3 (SEQ ID NO. 447). For each
human VH several degenerated oligonucleotides have to be
synthesized that overlap in a terminal stretch of approximately
15-20 nucleotides. To this end every second primer is an antisense
primer. For VH3 3-11 the following set of oligonucleotides is
used:
TABLE-US-00014 5'PM3-VH-A-XhoI (SEQ ID NO. 737) CCG GAT CTC GAG TCT
GGC GGC GGA CTG GTG AAG CCT GGC GRG TCC CTG ARG CTG TCC TGT
3'PM3-VH-B (SEQ ID NO. 738) CCA GTA CAT GTA GTA GTC GGA GAA GGT GAA
GCC GGA GGC GRY ACA GGA CAG CYT CAG GGA 5'PM3-VH-C (SEQ ID NO. 739)
TAC TAC ATG TAC TGG RTC CGC CAG RCC CCT GRG AAG SGG CTG GAA TGG GTG
KCC ATC ATC TCC GAC GGC 3'PM3-VH-D (SEQ ID NO. 740) GGC GTT GTC CCG
GGA GAT GGT GAA CCG GCC CTT GAT GAT GTC GGA GTA GTA GGT GTA GTA GCC
GCC GTC GGA GAT GAT 5'PM3-VH-E (SEQ ID NO. 741) TCC CGG GAC AAC GCC
AAG AAC ARC CTG TAC CTG CAG ATG ARC TCC CTG ARG KCC GAG GAC ACC GCC
RTG TAC TAC TGC RCC CGG GGC 3'PM3-VH-F-BstEII (SEQ ID NO. 742) CGA
TAC GGT GAC CAG GGT GCC CTG GCC CCA GTA ATC CAT GGC GCC GTG TCT CAG
CAG AGG GAA GCC CCG GGY GCA GTA GTA
[0619] For VH1 1-02 the oligonucleotides are as follows:
TABLE-US-00015 5'PM4-VH-A-XhoI (SEQ ID NO. 743) CTT GAT CTC GAG TCT
GGC GCC GAA STG RWG RAG CCT GGC GCC TCC GTG AAG STG TCC TGC AAG GCC
TCC GGC TAC 3'PM4-VH-B (SEQ ID NO. 744) CCA TTC CAG GCC CTG CYC AGG
CSY CTG CCG CAS CCA GTT GAT GTC GAA GTA GGT GAA GGT GTA GCC GGA GGC
CTT 5'PM4-VH-C (SEQ ID NO. 745) CAG GGC CTG GAA TGG ATS GGC GGC ATC
TCC CCT GGC GAC GGC AAC ACC AAC TAC AAC GAG AAC TTC AAG 3'PM4-VH-D
(SEQ ID NO. 746) AT GTA GGC GGT GGA GMT GGA CKT GTC TMT GGT CAK TGT
GRC CYT GCC CTT GAA GTT CTC GTT GTA 5'PM4-VH-E (SEQ ID NO. 747) C
TCC ACC GCC TAC ATS SAG CTG TCC CGG CTG ASA TCT GAS GAC ACC GCC GTG
TAC TWC TGC GCC AGG GAC GGC 3'PM4-VH-F-BstEII (SEQ ID NO. 748) AGA
CAC GGT CAC CGT GGT GCC CTG GCC CCA AGA GTC CAT GGC GTA GTA AGG GAA
GTT GCC GTC CCT GGC GCA
[0620] For VH1 1-03 the following oligonucleotides are used:
TABLE-US-00016 5'PM8-VH-A-XhoI (SEQ ID NO. 749) CTT GAT CTC GAG TCC
GGC SCT GAG STG RWG AAG CCT GGC GCC TCC GTG AAG RTG TCC TGC AAG GCC
TCC GGC TAC 3'PM8-VH-B (SEQ ID NO. 750) CCA TTC CAG CMS CTG GCC GGG
TKY CTG TYT CAC CCA GTG CAT CAC GTA GCC GGT GAA GGT GTA GCC GGA GGC
CTT GCA 5'PM8-VH-C (SEQ ID NO. 751) CCC GGC CAG SKG CTG GAA TGG ATS
GGC TAC ATC AAC CCT TAC AAC GAC GTG ACC CGG TAC AAC GGC AAG TTC AAG
3'PM8-VH-D (SEQ ID NO. 752) TTC CAT GTA GGC GGT GGA GGM GKA CKT GTC
KCT GGT AAK GGT GRC TYT GCC CTT GAA CTT GCC GTT GTA 5'PM8-VH-E (SEQ
ID NO. 753) TCC ACC GCC TAC ATG GAA CTG TCC RGC CTG ASG TCT GAG GAC
ACC GCC GTG TAC TAC TGC GCC AGG GGC 3'PM8-VH-F-BstEII (SEQ ID NO.
754) CGA TAC GGT GAC CAG AGT GCC TCT GCC CCA GGA GTC GAA GTA GTA
CCA GTT CTC GCC CCT GGC GCA GTA GTA
[0621] Each of these primer-sets spans over the whole corresponding
VH sequence. Within each set primers are mixed in equal amounts
(e.g. 1 .mu.l of each primer (primer stocks 20 to 100 .mu.M) to a
20 .mu.l PCR reaction) and added to a PCR mix consisting of PCR
buffer, nucleotides and Taq polymerase. This mix is incubated at
94.degree. C. for 3 minutes, 65.degree. C. for 1 minute, 62.degree.
C. for 1 minute, 59.degree. C. for 1 minute, 56.degree. C. for 1
minute, 52.degree. C. for 1 minute, 50.degree. C. for 1 minute and
at 72.degree. C. for 10 minutes in a PCR cycler. Subsequently the
product is run in an agarose gel electrophoresis and the product of
a size from 200 to 400 isolated from the gel according to standard
methods.
[0622] Each VH PCR product is then used as a template for a
standard PCR reaction using primers that incorporate N-terminal and
C-terminal suitable cloning restriction sites. The DNA fragment of
the correct size (for a VH approximately 350 nucleotides) is
isolated by agarose gel electrophoresis according to standard
methods. In this way sufficient VH DNA fragment is amplified.
[0623] The human antibody germline VL sequence VkI L1 (http
colon-slash-slash vbase.mrc-cpe.cam.ac.uk) is chosen as framework
context for CDRL1 (SEQ ID NO. 389), CDRL2 (SEQ ID NO. 390) and
CDRL3 (SEQ ID NO. 391). Likewise human antibody germline VL
sequence VkII A17 (http colon-slash-slash vbase.mrc-cpe.cam.ac.uk)
is chosen as framework context for CDRL1 (SEQ ID NO. 403), CDRL2
(SEQ ID NO. 404) and CDRL3 (SEQ ID NO. 405) as well as the human
antibody germline VL sequence VkII A1 (http colon-slash-slash
vbase.mrc-cpe.cam.ac.uk) as framework context for CDRL1 (SEQ ID NO.
450), CDRL2 (SEQ ID NO. 451) and CDRL3 (SEQ ID NO. 452). For each
human VL several degenerated oligonucleotides have to be
synthesized that overlap in a terminal stretch of approximately
15-20 nucleotides. To this end every second primer is an antisense
primer. Restriction sites needed for later cloning within the
oligonucleotides are deleted. For Vkl L1 the following
oligonucleotides are used:
TABLE-US-00017 5'PM3-VL-A-SacI (SEQ ID NO. 755) CTT GAT GAG CTC CAG
ATG ACC CAG TCC CCC ARS TYC MTG TCC RCC TCC GTG GGC GAC AGA GTG ACC
3'PM3-VL-B (SEQ ID NO. 756) GCC GGG CTT CTG CTG AWA CCA GGC CAC GTT
GGT GTC CAC GTT CTG GGA GGC CTT GCA GGT GAY GGT CAC TCT GTC GCC
5'PM3-VL-C (SEQ ID NO. 757) CAG CAG AAG CCC GGC MAG KCC CCT AAG KCC
CTG ATC TAC TCC GCC TCC TAC CGG TAC TCT 3'PM3-VL-D (SEQ ID NO. 758)
CAG GGT GAA GTC GGT GCC GGA CYC GGA GCC GGA GAA CCG GKM AGG CAC GYC
AGA GTA CCG GTA GGA 5'PM3-VL-E (SEQ ID NO. 759) ACC GAC TTC ACC CTG
ACC ATC TCC ARC STG CAG YCT GAG GAC YTC GCC RMG TAC TWC TGC CAG CAG
TAC GAC 3'PM3-VL-F-BsiWI/SpeI (SEQ ID NO. 760) CGA GTA ACT AGT CGT
ACG CTT GAT TTC CAG CTT GGT CCC TCC GCC GAA GGT GTA AGG GTA GGA GTC
GTA CTG CTG GCA
[0624] For VkII A17 the oligonucleotides are as follows:
TABLE-US-00018 5'PM4-VL-A-SacI (SEQ ID NO. 761) CTT GAT GAG CTC GTG
ATG ACC CAG TCC CCC CTG TCC CTG CCT GTG AYC CTG GGC SAM CMG GCC TCC
ATC TCC TGC CGG 3'PM4-VL-B (SEQ ID NO. 762) AAA CCA GTG CAG GTA GGT
ATT GCC GTT GGA GTG CAC CAG GGA CTG GGA GGA CCG GCA GGA GAT GGA GGC
5'PM4-VL-C (SEQ ID NO. 763) ACC TAC CTG CAC TGG TTT CWG CAG ARG CCT
GGC CAG TCC CCT ARG CKG CTG ATC TAC ACC GTG TCC AAC CGG 3'PM4-VL-D
(SEQ ID NO. 764) CAG GGT GAA GTC GGT GCC GGA GCC GGA GCC AGA GAA
CCT GTC AGG CAC GCC GGA GAA CCG GTT GGA CAC GGT 5'PM4-VL-E (SEQ ID
NO. 765) GGC ACC GAC TTC ACC CTG AAG ATC TCC CGG GTG GAG GCC GAA
GAT STG GGC GTG TAC TWT TGC TCC CAG TCC ACC 3'PM4-VL-F-BsiWI/SpeI
(SEQ ID NO. 766) ACT CAG ACT AGT CGT ACG CTT GAT TTC CAG CTT GGT
CCC TCC GCC GAA GGT AGG CAC GTG GGT GGA CTG GGA GCA
[0625] For VkII A1 the following oligonucleotides are used:
TABLE-US-00019 5'PM8-VL-A-SacI (SEQ ID NO. 767) CTT GAT GAG CTC GTG
ATG ACC CAG TCT CCA SYC TCC CTG SCT GTG ACT CTG GGC CAG CSG GCC TCC
ATC TCT TGC CGG 3'PM8-VL-B (SEQ ID NO. 768) CCA GTG CAT GAA GGT GTT
GTC GTA GGA GTC GAT GGA CTC GGA GGC CCG GCA AGA GAT GGA GGC
5'PM8-VL-C (SEQ ID NO. 769) ACC TTC ATG CAC TGG TWT CAG CAG ARG CCT
GGC CAG YCT CCT MRC CKG CTG ATC TWC CGG GCC TCT ATC CTG GAA
3'PM8-VL-D (SEQ ID NO. 770) CAG GGT GAA GTC GGT GCC GGA GCC AGA GCC
GGA GAA CCG GKC AGG GAY GCC GGA TTC CAG GAT AGA GGC CCG 5'PM8-VL-E
(SEQ ID NO. 771) ACC GAC TTC ACC CTG AMA ATC TMC CST GTG GAG GCC
GAS GAC GTG GSC RYC TAC TAC TGC CAC CAG 3'PM8-VL-F-BsiWI/SpeI (SEQ
ID NO. 772) ACT CAG ACT AGT CGT ACG CTT GAT TTC CAG CTT GGT CCC TCC
GCC GAA GGT GTA AGG GTC CTC GAT GGA CTG GTG GCA GTA GTA
[0626] Each of these primer-sets spans over the whole corresponding
VL sequence.
[0627] Within each set primers are mixed in equal amounts (e.g. 1
.mu.l of each primer (primer stocks 20 to 100 .mu.M) to a 20 .mu.l
PCR reaction) and added to a PCR mix consisting of PCR buffer,
nucleotides and Taq polymerase. This mix is incubated at 94.degree.
C. for 3 minutes, 65.degree. C. for 1 minute, 62.degree. C. for 1
minute, 59.degree. C. for 1 minute, 56.degree. C. for 1 minute,
52.degree. C. for 1 minute, 50.degree. C. for 1 minute and at
72.degree. C. for 10 minutes in a PCR cycler. Subsequently the
product is run in an agarose gel electrophoresis and the product of
a size from 200 to 400 isolated from the gel according to standard
methods.
[0628] Each VL PCR product is then used as a template for a
standard PCR reaction using primers that incorporate N-terminal and
C-terminal suitable cloning restriction sites. The DNA fragment of
the correct size (for a VL approximately 330 nucleotides) is
isolated by agarose gel electrophoresis according to standard
methods. In this way sufficient VL DNA fragment is amplified.
[0629] The final VH3 3-11-based VH PCR product (i.e. the repertoire
of human/humanized VH) is then combined with the final VkI L1-based
VL PCR product (i.e. the repertoire of human/humanized VL), the
final VH1 1-02-based VH PCR product (i.e. the repertoire of
human/humanized VH) is combined with the final VkII A17-based VL
PCR product (i.e. the repertoire of human/humanized VL) and the
final VH1 1-03-based VH PCR product (i.e. the repertoire of
human/humanized VH) is combined with the final VkII A1-based VL PCR
product (i.e. the repertoire of human/humanized VL) in the phage
display vector pComb3H5Bhis, respectively. These three VH-VL
combinations form three different libraries of functional scFvs
from which--after display on filamentous phage--anti-PSMA binders
are selected, screened, identified and confirmed as described in
the following:
[0630] 450 ng of the light chain fragments (SacI-SpeI digested) are
ligated with 1400 ng of the phagemid pComb3H5Bhis (SacI-SpeI
digested; large fragment). The resulting combinatorial antibody
library is then transformed into 300 ul of electrocompetent
Escherichia coli XL1 Blue cells by electroporation (2.5 kV, 0.2 cm
gap cuvette, 25 uFD, 200 Ohm, Biorad gene-pulser) resulting in a
library size of more than 10.sup.7 independent clones. After one
hour of phenotype expression, positive transformants are selected
for carbenicilline resistance encoded by the pComb3H5BHis vector in
100 ml of liquid super broth (SB)-culture over night. Cells are
then harvested by centrifugation and plasmid preparation is carried
out using a commercially available plasmid preparation kit
(Qiagen).
[0631] 2800 ng of this plasmid-DNA containing the VL-library
(Xhol-BstEII digested; large fragment) are ligated with 900 ng of
the heavy chain V-fragments (XhoI-BstEII digested) and again
transformed into two 300 ul aliquots of electrocompetent E. coli
XL1 Blue cells by electroporation (2.5 kV, 0.2 cm gap cuvette, 25
uFD, 200 Ohm) resulting in a total VH-VL scFv (single chain
variable fragment) library size of more than 10.sup.7 independent
clones.
[0632] After phenotype expression and slow adaptation to
carbenicilline, the E. coli cells containing the antibody library
are transferred into SB-carbenicilline (SB with 50 ug/mL
carbenicilline) selection medium. The E. coli cells containing the
antibody library is then infected with an infectious dose of
10.sup.12 particles of helper phage VCSM13 resulting in the
production and secretion of filamentous M13 phage, wherein phage
particle contains single stranded pComb3H5BHis-DNA encoding a
scFv-fragment and displayed the corresponding scFv-protein as a
translational fusion to phage coat protein III. This pool of phages
displaying the antibody library is used for the selection of
antigen binding entities.
[0633] For this purpose the phage library carrying the cloned
scFv-repertoire is harvested from the respective culture
supernatant by PEG8000/NaCl precipitation and centrifugation.
Approximately 10.sup.11to 10.sup.12 scFv phage particles are
resuspended in 0.4 ml of PBS/0.1% BSA and incubated with 10.sup.5
to 10.sup.7 PSMA-positive human prostate cancer cell line LNCaP
(ATCC No. CRL-1740) for 1 hour on ice under slow agitation. These
LNCaP cells are harvested beforehand by centrifugation, washed in
PBS and resuspended in PBS/1% FCS (containing 0.05% Na Azide). scFv
phage which do not specifically bind to LNCaP cells are eliminated
by up to five washing steps with PBS/1% FCS (containing 0.05% Na
Azide). After washing, binding entities are eluted from the cells
by resuspending the cells in HCl-glycine pH 2.2 (10 min incubation
with subsequent vortexing) and after neutralization with 2 M Tris
pH 12, the eluate is used for infection of a fresh uninfected E.
coli XL1 Blue culture (OD600>0.5). The E. coli culture
containing E. coli cells successfully transduced with a phagemid
copy, encoding a human/humanized scFv-fragment, are again selected
for carbenicilline resistance and subsequently infected with VCMS
13 helper phage to start the second round of antibody display and
in vitro selection. A total of 4 to 5 rounds of selections are
carried out, normally.
[0634] In order to screen for PSMA specific binders plasmid DNA
corresponding to 4 and 5 rounds of panning is isolated from E. coli
cultures after selection. For the production of soluble
scFv-protein, VH-VL-DNA fragments are excised from the plasmids
(XhoI-SpeI). These fragments are cloned via the same restriction
sites into the plasmid pComb3H5BFlag/His differing from the
original pComb3H5BHis in that the expression construct (e.g. scFv)
includes a Flag-tag (DYKDDDDK)(SEQ ID NO: 1054) between the scFv
and the His6-tag and the additional phage proteins are deleted.
After ligation, each pool (different rounds of panning) of plasmid
DNA is transformed into 100 .mu.l heat shock competent E. coli TG1
or XLI blue and plated onto carbenicilline LB-agar. Single colonies
are picked into 100 .mu.l of LB carb (50 ug/ml carbenicilline).
[0635] E. coli transformed with pComb3H5BFlag/His containing a
VL-and VH-segment produce soluble scFv in sufficient amounts after
induction with 1 mM IPTG. Due to a suitable signal sequence, the
scFv-chain is exported into the periplasma where it folds into a
functional conformation.
[0636] Single E. coli TG1 bacterial colonies from the
transformation plates are picked for periplasmic small scale
preparations and grown in SB-medium (e.g. 10 ml) supplemented with
20 mM MgCl.sub.2 and carbenicilline 50 .mu.g/ml (and re-dissolved
in PBS (e.g. 1 ml) after harvesting. By four rounds of freezing at
-70.degree. C. and thawing at 37.degree. C., the outer membrane of
the bacteria is destroyed by temperature shock and the soluble
periplasmic proteins including the scFvs are released into the
supernatant. After elimination of intact cells and cell-debris by
centrifugation, the supernatant containing the anti-PSMA scFvs is
collected and used for the identification of PSMA specific binders
as follows:
[0637] Binding of scFvs to PSMA is tested by flow cytometry on the
PSMA-positive human prostate cancer cell line LNCaP (ATCC No.
CRL-1740). A periplasmic small scale preparation as described above
without any grown bacteria is used as negative control.
[0638] For flow cytometry 2.5.times.10.sup.5 cells are incubated
with 50 ul of scFv periplasmic preparation or with 5 .mu.g/ml of
purified scFv in 50 .mu.l PBS with 2% FCS. The binding of scFv is
detected with an anti-His antibody (Penta-His Antibody, BSA free,
Qiagen GmbH, Hilden, FRG) at 2 .mu.g/ml in 50 .mu.l PBS with 2%
FCS. As a second step reagent a R-Phycoerythrin-conjugated affinity
purified F(ab')2 fragment, goat anti-mouse IgG (Fc-gamma fragment
specific), diluted 1:100 in 50 .mu.l PBS with 2% FCS (Dianova,
Hamburg, FRG) is used. The samples are measured on a FACSscan (BD
biosciences, Heidelberg, FRG).
[0639] Single clones are then analyzed for favourable properties
and amino acid sequence. PSMA specific scFvs are converted into
recombinant bispecific single chain antibodies by joining them via
a Gly.sub.4Ser.sub.1-linker with the CD3 specific scFv I2C (SEQ ID
185) or any other CD3 specific scFv of the invention to result in
constructs with the domain arrangement
VH.sub.PSMA-(Gly.sub.4Ser.sub.1).sub.3-Gly.sub.4Ser.sub.1-VH.sub.CD3-(Gly-
.sub.4Ser.sub.1).sub.3-VL.sub.CD3 or
VL.sub.PSMA-(Gly.sub.4Ser.sub.1).sub.3-VH.sub.PSMA-Gly.sub.4Ser.sub.1-VH.-
sub.CD3-(Gly.sub.4Ser.sub.1).sub.3-VL.sub.CD3 or alternative domain
arrangements. For expression in CHO cells the coding sequences of
(i) an N-terminal immunoglobulin heavy chain leader comprising a
start codon embedded within a Kozak consensus sequence and (ii) a
C-terminal His.sub.6-tag followed by a stop codon are both attached
in frame to the nucleotide sequence encoding the bispecific single
chain antibodies prior to insertion of the resulting DNA-fragment
as obtained by gene synthesis into the multiple cloning site of the
expression vector pEF-DHFR (Raum et al. Cancer Immunol Immunother
50 (2001) 141-150). Transfection of the generated expression
plasmids, protein expression and purification of cross-species
specific bispecific antibody constructs are performed as described
in chapters 24.6 and 24.7 of this example. All other state of the
art procedures are carried out according to standard protocols
(Sambrook, Molecular Cloning; A Laboratory Manual, 3rd edition,
Cold Spring Harbour Laboratory Press, Cold Spring Harbour, N.Y.
(2001)).
[0640] Identification of functional bispecific single-chain
antibody constructs is carried out by flow cytometric binding
analysis of culture supernatant from transfected cells expressing
the cross-species specific bispecific antibody constructs. The
flowcytometric analysis is perfomed on the human PSMA positive
prostate cancer cell line LNCaP (ATCC No. CRL-1740) as described in
chapter 24.7 of this example. Only those constructs showing
bispecific binding to human and macaque CD3 as well as to PSMA are
selected for further use.
[0641] Cytotoxic activity of the generated cross-species specific
bispecific single chain antibody constructs against PSMA positive
target cells elicited by effector T cells is analyzed as described
in chapter 24.8 of this example. The human PSMA positive prostate
cancer cell line LNCaP (ATCC No. CRL-1740) is used as source of
target cells. Only those constructs showing potent recruitment of
cytotoxic activity of effector T cells against target cells
positive for PSMA are selected for further use.
[0642] 25. Epitope Mapping of PSMA and CD3 Cross-Species Specific
Bispecific Single Chain Antibody Molecules
[0643] 25.1 Generation of CHO Cells Expressing Human/Rat PSMA
Chimeras
[0644] For mapping of the binding epitopes of PSMA cross-species
specific bispecific single chain antibody molecules, chimeric PSMA
proteins were generated with PSMA from two different species. This
approach requires that only the PSMA protein from one species is
recognized by the antibody. Here, PSMA of rattus norvegicus, which
is not bound by the tested PSMA cross-species specific bispecific
single chain antibody molecules, was used for making chimera with
human PSMA. Therefore creating a chimera in the region containing
the binding epitope of a PSMA cross-species specific bispecific
single chain antibody leads to loss of binding of said single chain
antibody to the respective PSMA construct.
[0645] The coding sequence of human PSMA as published in
GENBANK.RTM. (Accession number NM_004476) and the coding sequence
of rat PSMA (NM_057185, Rattus norvegicus folate hydrolase (Folh1),
mRNA, National Center for Biotechnology Information, http
colon-slash-slash www.ncbi.nlm.nih.gov/entrez) were used for
generation of the chimeric constructs.
[0646] A set of 7 chimeric cDNA constructs was designed and
generated by gene synthesis according to standard protocols. In the
constructs segments of the coding sequences for the amino acids 140
to 169, 191 to 258, 281 to 284, 300 to 344, 589 to 617, 683 to 690
and 716 to 750, respectively, were exchanged for the homologous
sequences of rat PSMA.
[0647] Chimeric PSMA constructs were generated as described above
and designated as set out in the following Table 9:
TABLE-US-00020 TABLE 9 Designation of chimeric PSMA constructs SEQ
ID (nucl/prot) Designation 1033/1034 huPSMArat140-169 1035/1036
huPSMArat191-258 1037/1038 huPSMArat281-284 1039/1040
huPSMArat300-344 1041/1042 huPSMArat598-617 1043/1044
huPSMArat683-690 1045/1046 huPSMArat716-750
[0648] The gene synthesis fragments were designed as to contain
first a Kozak site for eukaryotic expression of the construct
followed by the coding sequence of the chimeric PSMA proteins,
followed in frame by the coding sequence of a FLAG-tag and a stop
codon. The gene synthesis fragments were also designed as to
introduce restriction sites at the beginning and at the end of the
fragments. The introduced restriction sites, EcoRI at the 5' end
and SalI at the 3' end, were utilized in the following cloning
procedures. Undesirable internal restriction sites were removed by
silent mutation of the coding sequence in the gene synthesis
fragments. The gene synthesis fragments were cloned via EcoRI and
SalI into a plasmid designated pEF-DHFR (pEF-DHFR is described in
Raum et al. Cancer Immunol Immunother 50 (2001) 141-150) following
standard protocols. The aforementioned procedures were carried out
according to standard protocols (Sambrook, Molecular Cloning; A
Laboratory Manual, 3rd edition, Cold Spring Harbour Laboratory
Press, Cold Spring Harbour, N.Y. (2001)). A clone with
sequence-verified nucleotide sequence was transfected into DHFR
deficient CHO cells for eukaryotic expression of the construct.
Eukaryotic protein expression in DHFR deficient CHO cells was
performed as described by Kaufmann R. J. (1990) Methods Enzymol.
185, 537-566. Gene amplification of the construct was induced by
increasing concentrations of methotrexate (MTX) to a final
concentration of up to 20 nM MTX.
[0649] 25.2 Flow Cytometric Binding Analysis for Epitope Mapping of
PSMA and CD3 Cross-Species Specific Bispecific Single Chain
Antibody Molecules Using Chimeric PSMA Proteins
[0650] In order to determine the binding epitope of PSMA
cross-species specific bispecific single chain antibody constructs
a FACS analysis was performed. For this purpose CHO cells
transfected with human/rat chimeric PSMA molecules as described in
Example 25.1 were used. FACS analysis with supernatant of CHO cells
expressing bispecific single chain antibody constructs was
performed as described herein. Detection of binding of PSMA
cross-species specific bispecific single chain antibody constructs
was performed using a murine Penta His antibody and as second step
reagent an Fc gamma-specific antibody conjugated to phycoerythrin.
Supernatant of untransfected cells was used as a negative
control.
[0651] As shown in FIGS. 53A-53D all PSMA cross-species specific
bispecific single chain antibody constructs tested showed binding
to the chimeric constructs huPSMArat140-169, huPSMArat191-258,
huPSMArat281-284, huPSMArat683-690 and huPSMArat716-750. As
furthermore shown in FIGS. 53A-53D there is a lack of binding for
the PSMA cross-species specific bispecific single chain antibody
constructs PM84-D7.times.I2C, PM29-G1.times.I2C and
PM49-B9.times.I2C to the construct huPSMArat300-344, which
demonstrates the presence of a major binding epitope for these
constructs in the region of amino acids 300 to 344 of human PSMA.
As also shown in FIGS. 53A-53D there is a lack of binding for the
PSMA cross-species specific bispecific single chain antibody
construct PM34-C7.times.I2C to the construct huPSMArat598-617,
which demonstrates the presence of a major binding epitope for this
construct in the region of amino acids 598 to 617 of human
PSMA.
[0652] 26 Epitope Mapping Using a Peptide Scanning Approach
[0653] The two PSMA BiTE antibodies PM 76-610.times.I2C and PM
76-A9.times.I2C were cross-reactive with rat PSMA, which excluded
them from mapping by using human-rat PSMA chimeras. Likewise,
binding signals of PSMA BiTE antibody PM F1-A10.times.I2C on
human-rat PSMA chimeras were too weak for reliable epitope mapping.
These three PSMA BiTE antibodies were subjected to an alternative
epitope mapping approach based on peptide scanning (Pepscan).
Pepscan uses overlapping peptides of a given protein and analyses
antibody binding to immobilized peptides by enzyme-linked
immunosorbent assays (ELISAs). The epitope mapping experiments with
PSMA BiTE antibodies were performed at the company Pepscan
(Lelystad, The Netherlands). A detailed description of the method
is found elsewhere (Bernard et al. 2004, J. Biol. Chem., 279:
24313-22; Teeling et al. 2006, J Immunol., 177: 362-71).
[0654] In brief, 693 different 15-mer peptides were synthesized
that span the entire extracellular amino acid sequence of human
PSMA and overlap with each neighbouring 15-mer peptide by 14 amino
acids. These peptides were coated to ELISA wells in a 384-well
plate format. For this series of experiments, anti-PSMA scFvs of
the respective BiTE antibody candidates (scFv MP 9076-A9 for BiTE
antibody PM 76-A9.times.I2C; scFv MP 9076-610 for BiTE antibody PM
76-610.times.I2C; scFv F1-A10 for BiTE antibody PM
F1-A10.times.I2C) were produced in E. coli and used for ELISA as
crude periplasmic extracts. To this end 7 ml of crude periplasmic
extracts were shipped on dry ice to Pepscan (The Netherlands).
Using scFv counterparts in this assay minimized the risk to pick up
signals from the second non-PSMA binding specificity of the BiTE
antibodies, which may lead to misinterpretation of the PSMA binding
epitopes of the target binders. The scFvs were incubated with the
peptides and specific binding detected using an anti-His antibody.
Binding signals were measured in a 384-well ELISA reader. Results
are shown in FIGS. 54, 55 and 56.
[0655] Of the three anti-PSMA scFv antibodies used to generate PSMA
BiTE antibodies (scFv MP 9076-A9 for BiTE antibody PM
76-A9.times.I2C; scFv MP 9076-610 for BiTE antibody PM
76-610.times.I2C; scFv F1-A10 for BiTE antibody PM
F1-A10.times.I2C) apparently two (MP 9076-A9 and MP 9076-610) bound
to a similar dominant epitope of human PSMA. This finding is
supported by the close homology of the two scFv antibodies and
sequence identity in their six CDRs. The peptide binding signals
point to a core epitope between Thr334 to Thr339. This sequence is
located in an exposed loop of the apical domain of human PSMA as is
shown in FIG. 57. For scFv F1-A10, a dominant epitope could be
detected within the sequence LFEPPPPGYENVS (amino acids 143-155 of
human PSMA), which is also localized in the apical domain. The
strong binding of the three antibody fragments MP 9076-A9.
MP9076-610 and F1-A10 to discrete peptides indicates recognition of
a linear protein epitope rather than a carbohydrate moiety.
TABLE-US-00021 SEQ ID NO. DESIGNATION SOURCE TYPE SEQUENCE 1. Human
CD3.epsilon. extracellular domain human aa
QDGNEEMGGITQTPYKVSISGTTVILTCPQYPGSEILWQHNDKNIGGDEDDKNIGSDEDHLSLKE
FSELEQSGYYVCYPRGSKPEDANFYLYLRARVCENCMEMD 2. Human CD3.epsilon. 1-27
human aa QDGNEEMGGITQTPYKVSISGTTVILT 3. Callithrix jacchus
CD3.epsilon. extracellular domain Callithrix aa
QDGNEEMGDTTQNPYKVSISGTTVTLTCPRYDGHEIKWLVNSQNKEGHEDHLLLEDFSEMEQSGY
jacchus YACLSKETPAEEASHYLYLKARVCENCVEVD 4. Callithrix jacchus
CD3.epsilon. 1-27 Callithrix aa QDGNEEMGDTTQNPYKVSISGTTVTLT jacchus
5. Saguinus oedipus CD3.epsilon. extracellular domain Saguinus aa
QDGNEEMGDTTQNPYKVSISGTTVTLTCPRYDGHEIKWLVNSQNKEGHEDHLLLEDFSEMEQSGY
oedipus YACLSKETPAEEASHYLYLKARVCENCVEVD 6. Saguinus oedipus
CD3.epsilon. 1-27 Saguinus aa QDGNEEMGDTTQNPYKVSISGTTVTLT oedipus
7. Saimiri sciureus CD3.epsilon. extracellular domain Saimiri aa
QDGNEEIGDTTQNPYKVSISGTTVTLTCPRYDGQEIKWLVNDQNKEGHEDHLLLEDFSEMEQSGY
sciureus YACLSKETPTEEASHYLYLKARVCENCVEVD 8. Saimiri sciureus
CD3.epsilon. 1-27 Saimiri aa QDGNEEIGDTTQNPYKVSISGTTVTLT sciureus
9. CDR-L1 of F6A artificial aa GSSTGAVTSGYYPN 10. CDR-L2 of F6A
artificial aa GTKFLAP 11. CDR-L3 of F6A artificial aa ALWYSNRWV 12.
CDR-H1 of F6A artificial aa IYAMN 13. CDR-H2 of F6A artificial aa
RIRSKYNNYATYYADSVKS 14. CDR-H3 of F6A artificial aa HGNFGNSYVSFFAY
15. VH of F6A artificial aa
EVQLVESGGGLVQPGGSLKLSCAASGFTFNIYAMNWVRQAPGKGLEWVARIRSKYNNYATYYADS
VKSRFTISRDDSKNTAYLQMNNLKTEDTAVYYCVRHGNFGNSYVSFFAYWGQGTLVTVSS 16. VH
of F6A artificial nt
GAGGTGCAGCTGGTCGAGTCTGGAGGAGGATTGGTGCAGCCTGGAGGGTCATTGAAACTCTCATG
TGCAGCCTCTGGATTCACCTTCAATATCTACGCCATGAACTGGGTCCGCCAGGCTCCAGGAAAGG
GTTTGGAATGGGTTGCTCGCATAAGAAGTAAATATAATAATTATGCAACATATTATGCCGATTCA
GTGAAAAGCAGGTTCACCATCTCCAGAGATGATTCAAAAAACACTGCCTATCTACAAATGAACAA
CTTGAAAACTGAGGACACTGCCGTGTACTACTGTGTGAGACATGGGAACTTCGGTAATAGCTACG
TATCCTTCTTCGCTTACTGGGGCCAAGGGACTCTGGTCACCGTCTCCTCA 17. VL of F6A
artificial aa
QTVVTQEPSLTVSPGGTVTLTCGSSTGAVTSGYYPNWVQQKPGQAPRGLIGGTKFLAPGTPARFS
GSLLGGKAALTLSGVQPEDEAEYYCALWYSNRWVFGGGTKLTVL 18. VL of F6A
artificial nt
CAGACTGTTGTGACTCAGGAACCTTCACTCACCGTATCACCTGGTGGAACAGTCACACTCACTTG
TGGCTCCTCGACTGGGGCTGTTACATCTGGCTACTACCCAAACTGGGTCCAACAAAAACCAGGTC
AGGCACCCCGTGGTCTAATAGGTGGGACTAAGTTCCTCGCCCCCGGTACTCCTGCCAGATTCTCA
GGCTCCCTGCTTGGAGGCAAGGCTGCCCTCACCCTCTCAGGGGTACAGCCAGAGGATGAGGCAGA
ATATTACTGTGCTCTATGGTACAGCAACCGCTGGGTGTTCGGTGGAGGAACCAAACTGACTGTCC
TA 19. VH-P of F6A artificial aa
EVQLLESGGGLVQPGGSLKLSCAASGFTFNIYAMNWVRQAPGKGLEWVARIRSKYNNYATYYADS
VKSRFTISRDDSKNTAYLQMNNLKTEDTAVYYCVRHGNFGNSYVSFFAYWGQGTLVTVSS 20.
VH-P of F6A artificial nt
GAGGTGCAGCTGCTCGAGTCTGGAGGAGGATTGGTGCAGCCTGGAGGGTCATTGAAACTCTCATG
TGCAGCCTCTGGATTCACCTTCAATATCTACGCCATGAACTGGGTCCGCCAGGCTCCAGGAAAGG
GTTTGGAATGGGTTGCTCGCATAAGAAGTAAATATAATAATTATGCAACATATTATGCCGATTCA
GTGAAAAGCAGGTTCACCATCTCCAGAGATGATTCAAAAAACACTGCCTATCTACAAATGAACAA
CTTGAAAACTGAGGACACTGCCGTGTACTACTGTGTGAGACATGGGAACTTCGGTAATAGCTACG
TATCCTTCTTCGCTTACTGGGGCCAAGGGACTCTGGTCACCGTCTCCTCA 21. VL-P of F6A
artificial aa
ELVVTQEPSLTVSPGGTVTLTCGSSTGAVTSGYYPNWVQQKPGQAPRGLIGGTKFLAPGTPARFS
GSLLGGKAALTLSGVQPEDEAEYYCALWYSNRWVFGGGTKLTVL 22. VL-P of F6A
artificial nt
GAGCTCGTTGTGACTCAGGAACCTTCACTCACCGTATCACCTGGTGGAACAGTCACACTCACTTG
TGGCTCCTCGACTGGGGCTGTTACATCTGGCTACTACCCAAACTGGGTCCAACAAAAACCAGGTC
AGGCACCCCGTGGTCTAATAGGTGGGACTAAGTTCCTCGCCCCCGGTACTCCTGCCAGATTCTCA
GGCTCCCTGCTTGGAGGCAAGGCTGCCCTCACCCTCTCAGGGGTACAGCCAGAGGATGAGGCAGA
ATATTACTGTGCTCTATGGTACAGCAACCGCTGGGTGTTCGGTGGAGGAACCAAACTGACTGTCC
TA 23. VH-VL of F6A artificial aa
EVQLVESGGGLVQPGGSLKLSCAASGFTFNIYAMNWVRQAPGKGLEWVARIRSKYNNYATYYADS
VKSRFTISRDDSKNTAYLQMNNLKTEDTAVYYCVRHGNFGNSYVSFFAYWGQGTLVTVSSGGGGS
GGGGSGGGGSQTVVTQEPSLTVSPGGTVTLTCGSSTGAVTSGYYPNWVQQKPGQAPRGLIGGTKF
LAPGTPARFSGSLLGGKAALTLSGVQPEDEAEYYCALWYSNRWVFGGGTKLTVL 24. VH-VL of
F6A artificial nt
GAGGTGCAGCTGGTCGAGTCTGGAGGAGGATTGGTGCAGCCTGGAGGGTCATTGAAACTCTCATG
TGCAGCCTCTGGATTCACCTTCAATATCTACGCCATGAACTGGGTCCGCCAGGCTCCAGGAAAGG
GTTTGGAATGGGTTGCTCGCATAAGAAGTAAATATAATAATTATGCAACATATTATGCCGATTCA
GTGAAAAGCAGGTTCACCATCTCCAGAGATGATTCAAAAAACACTGCCTATCTACAAATGAACAA
CTTGAAAACTGAGGACACTGCCGTGTACTACTGTGTGAGACATGGGAACTTCGGTAATAGCTACG
TATCCTTCTTCGCTTACTGGGGCCAAGGGACTCTGGTCACCGTCTCCTCAGGTGGTGGTGGTTCT
GGCGGCGGCGGCTCCGGTGGTGGTGGTTCTCAGACTGTTGTGACTCAGGAACCTTCACTCACCGT
ATCACCTGGTGGAACAGTCACACTCACTTGTGGCTCCTCGACTGGGGCTGTTACATCTGGCTACT
ACCCAAACTGGGTCCAACAAAAACCAGGTCAGGCACCCCGTGGTCTAATAGGTGGGACTAAGTTC
CTCGCCCCCGGTACTCCTGCCAGATTCTCAGGCTCCCTGCTTGGAGGCAAGGCTGCCCTCACCCT
CTCAGGGGTACAGCCAGAGGATGAGGCAGAATATTACTGTGCTCTATGGTACAGCAACCGCTGGG
TGTTCGGTGGAGGAACCAAACTGACTGTCCTA 25. VH-VL-P of F6A artificial aa
EVQLLESGGGLVQPGGSLKLSCAASGFTFNIYAMNWVRQAPGKGLEWVARIRSKYNNYATYYADS
VKSRFTISRDDSKNTAYLQMNNLKTEDTAVYYCVRHGNFGNSYVSFFAYWGQGTLVTVSSGGGGS
GGGGSGGGGSELVVTQEPSLTVSPGGTVTLTCGSSTGAVTSGYYPNWVQQKPGQAPRGLIGGTKF
LAPGTPARFSGSLLGGKAALTLSGVQPEDEAEYYCALWYSNRWVFGGGTKLTVL 26. VH-VL-P
of F6A artificial nt
GAGGTGCAGCTGCTCGAGTCTGGAGGAGGATTGGTGCAGCCTGGAGGGTCATTGAAACTCTCATG
TGCAGCCTCTGGATTCACCTTCAATATCTACGCCATGAACTGGGTCCGCCAGGCTCCAGGAAAGG
GTTTGGAATGGGTTGCTCGCATAAGAAGTAAATATAATAATTATGCAACATATTATGCCGATTCA
GTGAAAAGCAGGTTCACCATCTCCAGAGATGATTCAAAAAACACTGCCTATCTACAAATGAACAA
CTTGAAAACTGAGGACACTGCCGTGTACTACTGTGTGAGACATGGGAACTTCGGTAATAGCTACG
TATCCTTCTTCGCTTACTGGGGCCAAGGGACTCTGGTCACCGTCTCCTCAGGTGGTGGTGGTTCT
GGCGGCGGCGGCTCCGGTGGTGGTGGTTCTGAGCTCGTTGTGACTCAGGAACCTTCACTCACCGT
ATCACCTGGTGGAACAGTCACACTCACTTGTGGCTCCTCGACTGGGGCTGTTACATCTGGCTACT
ACCCAAACTGGGTCCAACAAAAACCAGGTCAGGCACCCCGTGGTCTAATAGGTGGGACTAAGTTC
CTCGCCCCCGGTACTCCTGCCAGATTCTCAGGCTCCCTGCTTGGAGGCAAGGCTGCCCTCACCCT
CTCAGGGGTACAGCCAGAGGATGAGGCAGAATATTACTGTGCTCTATGGTACAGCAACCGCTGGG
TGTTCGGTGGAGGAACCAAACTGACTGTCCTA 27. CDR-L1 of H2C artificial aa
GSSTGAVTSGYYPN 28. CDR-L2 of H2C artificial aa GTKFLAP 29. CDR-L3
of H2C artificial aa ALWYSNRWV 30. CDR-H1 of H2C artificial aa
KYAMN 31. CDR-H2 of H2C artificial aa RIRSKYNNYATYYADSVKD 32.
CDR-H3 of H2C artificial aa HGNFGNSYISYWAY 33. VH of H2C artificial
aa
EVQLVESGGGLVQPGGSLKLSCAASGFTFNKYAMNWVRQAPGKGLEWVARIRSKYNNYATYYADS
VKDRFTISRDDSKNTAYLQMNNLKTEDTAVYYCVRHGNFGNSYISYWAYWGQGTLVTVSS 34. VH
of H2C artificial nt
GAGGTGCAGCTGGTCGAGTCTGGAGGAGGATTGGTGCAGCCTGGAGGGTCATTGAAACTCTCATG
TGCAGCCTCTGGATTCACCTTCAATAAGTACGCCATGAACTGGGTCCGCCAGGCTCCAGGAAAGG
GTTTGGAATGGGTTGCTCGCATAAGAAGTAAATATAATAATTATGCAACATATTATGCCGATTCA
GTGAAAGACAGGTTCACCATCTCCAGAGATGATTCAAAAAACACTGCCTATCTACAAATGAACAA
CTTGAAAACTGAGGACACTGCCGTGTACTACTGTGTGAGACATGGGAACTTCGGTAATAGCTACA
TATCCTACTGGGCTTACTGGGGCCAAGGGACTCTGGTCACCGTCTCCTCA 35. VL of H2C
artificial aa
QTVVTQEPSLTVSPGGTVTLTCGSSTGAVTSGYYPNWVQQKPGQAPRGLIGGTKFLAPGTPARFS
GSLLGGKAALTLSGVQPEDEAEYYCALWYSNRWVFGGGTKLTVL 36. VL of H2C
artificial nt
CAGACTGTTGTGACTCAGGAACCTTCACTCACCGTATCACCTGGTGGAACAGTCACACTCACTTG
TGGCTCCTCGACTGGGGCTGTTACATCTGGCTACTACCCAAACTGGGTCCAACAAAAACCAGGTC
AGGCACCCCGTGGTCTAATAGGTGGGACTAAGTTCCTCGCCCCCGGTACTCCTGCCAGATTCTCA
GGCTCCCTGCTTGGAGGCAAGGCTGCCCTCACCCTCTCAGGGGTACAGCCAGAGGATGAGGCAGA
ATATTACTGTGCTCTATGGTACAGCAACCGCTGGGTGTTCGGTGGAGGAACCAAACTGACTGTCC
TA 37. VH-P of H2C artificial aa
EVQLLESGGGLVQPGGSLKLSCAASGFTFNKYAMNWVRQAPGKGLEWVARIRSKYNNYATYYADS
VKDRFTISRDDSKNTAYLQMNNLKTEDTAVYYCVRHGNFGNSYISYWAYWGQGTLVTVSS 38.
VH-P of H2C artificial nt
GAGGTGCAGCTGCTCGAGTCTGGAGGAGGATTGGTGCAGCCTGGAGGGTCATTGAAACTCTCATG
TGCAGCCTCTGGATTCACCTTCAATAAGTACGCCATGAACTGGGTCCGCCAGGCTCCAGGAAAGG
GTTTGGAATGGGTTGCTCGCATAAGAAGTAAATATAATAATTATGCAACATATTATGCCGATTCA
GTGAAAGACAGGTTCACCATCTCCAGAGATGATTCAAAAAACACTGCCTATCTACAAATGAACAA
CTTGAAAACTGAGGACACTGCCGTGTACTACTGTGTGAGACATGGGAACTTCGGTAATAGCTACA
TATCCTACTGGGCTTACTGGGGCCAAGGGACTCTGGTCACCGTCTCCTCA 39. VL-P of H2C
artificial aa
ELVVTQEPSLTVSPGGTVTLTCGSSTGAVTSGYYPNWVQQKPGQAPRGLIGGTKFLAPGTPARFS
GSLLGGKAALTLSGVQPEDEAEYYCALWYSNRWVFGGGTKLTVL 40. VL-P of H2C
artificial nt
GAGCTCGTTGTGACTCAGGAACCTTCACTCACCGTATCACCTGGTGGAACAGTCACACTCACTTG
TGGCTCCTCGACTGGGGCTGTTACATCTGGCTACTACCCAAACTGGGTCCAACAAAAACCAGGTC
AGGCACCCCGTGGTCTAATAGGTGGGACTAAGTTCCTCGCCCCCGGTACTCCTGCCAGATTCTCA
GGCTCCCTGCTTGGAGGCAAGGCTGCCCTCACCCTCTCAGGGGTACAGCCAGAGGATGAGGCAGA
ATATTACTGTGCTCTATGGTACAGCAACCGCTGGGTGTTCGGTGGAGGAACCAAACTGACTGTCC
TA 41. VH-VL of H2C artificial aa
EVQLVESGGGLVQPGGSLKLSCAASGFTFNKYAMNWVRQAPGKGLEWVARIRSKYNNYATYYADS
VKDRFTISRDDSKNTAYLQMNNLKTEDTAVYYCVRHGNFGNSYISYWAYWGQGTLVTVSSGGGGS
GGGGSGGGGSQTVVTQEPSLTVSPGGTVTLTCGSSTGAVTSGYYPNWVQQKPGQAPRGLIGGTKF
LAPGTPARFSGSLLGGKAALTLSGVQPEDEAEYYCALWYSNRWVFGGGTKLTVL 42. VH-VL of
H2C artificial nt
GAGGTGCAGCTGGTCGAGTCTGGAGGAGGATTGGTGCAGCCTGGAGGGTCATTGAAACTCTCATG
TGCAGCCTCTGGATTCACCTTCAATAAGTACGCCATGAACTGGGTCCGCCAGGCTCCAGGAAAGG
GTTTGGAATGGGTTGCTCGCATAAGAAGTAAATATAATAATTATGCAACATATTATGCCGATTCA
GTGAAAGACAGGTTCACCATCTCCAGAGATGATTCAAAAAACACTGCCTATCTACAAATGAACAA
CTTGAAAACTGAGGACACTGCCGTGTACTACTGTGTGAGACATGGGAACTTCGGTAATAGCTACA
TATCCTACTGGGCTTACTGGGGCCAAGGGACTCTGGTCACCGTCTCCTCAGGTGGTGGTGGTTCT
GGCGGCGGCGGCTCCGGTGGTGGTGGTTCTCAGACTGTTGTGACTCAGGAACCTTCACTCACCGT
ATCACCTGGTGGAACAGTCACACTCACTTGTGGCTCCTCGACTGGGGCTGTTACATCTGGCTACT
ACCCAAACTGGGTCCAACAAAAACCAGGTCAGGCACCCCGTGGTCTAATAGGTGGGACTAAGTTC
CTCGCCCCCGGTACTCCTGCCAGATTCTCAGGCTCCCTGCTTGGAGGCAAGGCTGCCCTCACCCT
CTCAGGGGTACAGCCAGAGGATGAGGCAGAATATTACTGTGCTCTATGGTACAGCAACCGCTGGG
TGTTCGGTGGAGGAACCAAACTGACTGTCCTA 43. VH-VL-P of H2C artificial aa
EVQLLESGGGLVQPGGSLKLSCAASGFTFNKYAMNWVRQAPGKGLEWVARIRSKYNNYATYYADS
VKDRFTISRDDSKNTAYLQMNNLKTEDTAVYYCVRHGNFGNSYISYWAYWGQGTLVTVSSGGGGS
GGGGSGGGGSELVVTQEPSLTVSPGGTVTLTCGSSTGAVTSGYYPNWVQQKPGQAPRGLIGGTKF
LAPGTPARFSGSLLGGKAALTLSGVQPEDEAEYYCALWYSNRWVFGGGTKLTVL 44. VH-VL-P
of H2C artificial nt
GAGGTGCAGCTGCTCGAGTCTGGAGGAGGATTGGTGCAGCCTGGAGGGTCATTGAAACTCTCATG
TGCAGCCTCTGGATTCACCTTCAATAAGTACGCCATGAACTGGGTCCGCCAGGCTCCAGGAAAGG
GTTTGGAATGGGTTGCTCGCATAAGAAGTAAATATAATAATTATGCAACATATTATGCCGATTCA
GTGAAAGACAGGTTCACCATCTCCAGAGATGATTCAAAAAACACTGCCTATCTACAAATGAACAA
CTTGAAAACTGAGGACACTGCCGTGTACTACTGTGTGAGACATGGGAACTTCGGTAATAGCTACA
TATCCTACTGGGCTTACTGGGGCCAAGGGACTCTGGTCACCGTCTCCTCAGGTGGTGGTGGTTCT
GGCGGCGGCGGCTCCGGTGGTGGTGGTTCTGAGCTCGTTGTGACTCAGGAACCTTCACTCACCGT
ATCACCTGGTGGAACAGTCACACTCACTTGTGGCTCCTCGACTGGGGCTGTTACATCTGGCTACT
ACCCAAACTGGGTCCAACAAAAACCAGGTCAGGCACCCCGTGGTCTAATAGGTGGGACTAAGTTC
CTCGCCCCCGGTACTCCTGCCAGATTCTCAGGCTCCCTGCTTGGAGGCAAGGCTGCCCTCACCCT
CTCAGGGGTACAGCCAGAGGATGAGGCAGAATATTACTGTGCTCTATGGTACAGCAACCGCTGGG
TGTTCGGTGGAGGAACCAAACTGACTGTCCTA 45. CDR-L1 of H1E artificial aa
GSSTGAVTSGYYPN 46. CDR-L2 of H1E artificial aa GTKFLAP 47. CDR-L3
of H1E artificial aa ALWYSNRWV 48. CDR-H1 of H1E artificial aa
SYAMN 49. CDR-H2 of H1E artificial aa RIRSKYNNYATYYADSVKG 50.
CDR-H3 of H1E artificial aa HGNFGNSYLSFWAY 51. VH of H1E artificial
aa
EVQLVESGGGLEQPGGSLKLSCAASGFTFNSYAMNWVRQAPGKGLEWVARIRSKYNNYATYYADS
VKGRFTISRDDSKNTAYLQMNNLKTEDTAVYYCVRHGNFGNSYLSFWAYWGQGTLVTVSS
52. VH of H1E artificial nt
GAGGTGCAGCTGGTCGAGTCTGGAGGAGGATTGGAGCAGCCTGGAGGGTCATTGAAACTCTCATG
TGCAGCCTCTGGATTCACCTTCAATTCGTACGCCATGAACTGGGTCCGCCAGGCTCCAGGAAAGG
GTTTGGAATGGGTTGCTCGCATAAGAAGTAAATATAATAATTATGCAACATATTATGCCGATTCA
GTGAAAGGGAGGTTCACCATCTCCAGAGATGATTCAAAAAACACTGCCTATCTACAAATGAACAA
CTTGAAAACTGAGGACACTGCCGTGTACTACTGTGTGAGACATGGGAACTTCGGTAATAGCTACC
TATCCTTCTGGGCTTACTGGGGCCAAGGGACTCTGGTCACCGTCTCCTC 53. VL of H1E
artificial aa
QTVVTQEPSLTVSPGGTVTLTCGSSTGAVTSGYYPNWVQQKPGQAPRGLIGGTKFLAPGTPARFS
GSLLGGKAALTLSGVQPEDEAEYYCALWYSNRWVFGGGTKLTVL 54. VL of H1E
artificial nt
CAGACTGTTGTGACTCAGGAACCTTCACTCACCGTATCACCTGGTGGAACAGTCACACTCACTTG
TGGCTCCTCGACTGGGGCTGTTACATCTGGCTACTACCCAAACTGGGTCCAACAAAAACCAGGTC
AGGCACCCCGTGGTCTAATAGGTGGGACTAAGTTCCTCGCCCCCGGTACTCCTGCCAGATTCTCA
GGCTCCCTGCTTGGAGGCAAGGCTGCCCTCACCCTCTCAGGGGTACAGCCAGAGGATGAGGCAGA
ATATTACTGTGCTCTATGGTACAGCAACCGCTGGGTGTTCGGTGGAGGAACCAAACTGACTGTCC
TA 55. VH-P of H1E artificial aa
EVQLLESGGGLEQPGGSLKLSCAASGFTFNSYAMNWVRQAPGKGLEWVARIRSKYNNYATYYADS
VKGRFTISRDDSKNTAYLQMNNLKTEDTAVYYCVRHGNFGNSYLSFWAYWGQGTLVTVSS 56.
VH-P of H1E artificial nt
GAGGTGCAGCTGCTCGAGTCTGGAGGAGGATTGGAGCAGCCTGGAGGGTCATTGAAACTCTCATG
TGCAGCCTCTGGATTCACCTTCAATTCGTACGCCATGAACTGGGTCCGCCAGGCTCCAGGAAAGG
GTTTGGAATGGGTTGCTCGCATAAGAAGTAAATATAATAATTATGCAACATATTATGCCGATTCA
GTGAAAGGGAGGTTCACCATCTCCAGAGATGATTCAAAAAACACTGCCTATCTACAAATGAACAA
CTTGAAAACTGAGGACACTGCCGTGTACTACTGTGTGAGACATGGGAACTTCGGTAATAGCTACC
TATCCTTCTGGGCTTACTGGGGCCAAGGGACTCTGGTCACCGTCTCCTCA 57. VL-P of H1E
artificial aa
ELVVTQEPSLTVSPGGTVTLTCGSSTGAVTSGYYPNWVQQKPGQAPRGLIGGTKFLAPGTPARFS
GSLLGGKAALTLSGVQPEDEAEYYCALWYSNRWVFGGGTKLTVL 58. VL-P of H1E
artificial nt
GAGCTCGTTGTGACTCAGGAACCTTCACTCACCGTATCACCTGGTGGAACAGTCACACTCACTTG
TGGCTCCTCGACTGGGGCTGTTACATCTGGCTACTACCCAAACTGGGTCCAACAAAAACCAGGTC
AGGCACCCCGTGGTCTAATAGGTGGGACTAAGTTCCTCGCCCCCGGTACTCCTGCCAGATTCTCA
GGCTCCCTGCTTGGAGGCAAGGCTGCCCTCACCCTCTCAGGGGTACAGCCAGAGGATGAGGCAGA
ATATTACTGTGCTCTATGGTACAGCAACCGCTGGGTGTTCGGTGGAGGAACCAAACTGACTGTCC
TA 59. VH-VL of H1E artificial aa
EVQLVESGGGLEQPGGSLKLSCAASGFTFNSYAMNWVRQAPGKGLEWVARIRSKYNNYATYYADS
VKGRFTISRDDSKNTAYLQMNNLKTEDTAVYYCVRHGNFGNSYLSFWAYWGQGTLVTVSSGGGGS
GGGGSGGGGSQTVVTQEPSLTVSPGGTVTLTCGSSTGAVTSGYYPNWVQQKPGQAPRGLIGGTKF
LAPGTPARFSGSLLGGKAALTLSGVQPEDEAEYYCALWYSNRWVFGGGTKLTVL 60. VH-VL of
H1E artificial nt
GAGGTGCAGCTGGTCGAGTCTGGAGGAGGATTGGAGCAGCCTGGAGGGTCATTGAAACTCTCATG
TGCAGCCTCTGGATTCACCTTCAATTCGTACGCCATGAACTGGGTCCGCCAGGCTCCAGGAAAGG
GTTTGGAATGGGTTGCTCGCATAAGAAGTAAATATAATAATTATGCAACATATTATGCCGATTCA
GTGAAAGGGAGGTTCACCATCTCCAGAGATGATTCAAAAAACACTGCCTATCTACAAATGAACAA
CTTGAAAACTGAGGACACTGCCGTGTACTACTGTGTGAGACATGGGAACTTCGGTAATAGCTACC
TATCCTTCTGGGCTTACTGGGGCCAAGGGACTCTGGTCACCGTCTCCTCAGGTGGTGGTGGTTCT
GGCGGCGGCGGCTCCGGTGGTGGTGGTTCTCAGACTGTTGTGACTCAGGAACCTTCACTCACCGT
ATCACCTGGTGGAACAGTCACACTCACTTGTGGCTCCTCGACTGGGGCTGTTACATCTGGCTACT
ACCCAAACTGGGTCCAACAAAAACCAGGTCAGGCACCCCGTGGTCTAATAGGTGGGACTAAGTTC
CTCGCCCCCGGTACTCCTGCCAGATTCTCAGGCTCCCTGCTTGGAGGCAAGGCTGCCCTCACCCT
CTCAGGGGTACAGCCAGAGGATGAGGCAGAATATTACTGTGCTCTATGGTACAGCAACCGCTGGG
TGTTCGGTGGAGGAACCAAACTGACTGTCCTA 61. VH-VL-P of H1E artificial aa
EVQLLESGGGLEQPGGSLKLSCAASGFTFNSYAMNWVRQAPGKGLEWVARIRSKYNNYATYYADS
VKGRFTISRDDSKNTAYLQMNNLKTEDTAVYYCVRHGNFGNSYLSFWAYWGQGTLVTVSSGGGGS
GGGGSGGGGSELVVTQEPSLTVSPGGTVTLTCGSSTGAVTSGYYPNWVQQKPGQAPRGLIGGTKF
LAPGTPARFSGSLLGGKAALTLSGVQPEDEAEYYCALWYSNRWVFGGGTKLTVL 62. VH-VL-P
of H1E artificial nt
GAGGTGCAGCTGCTCGAGTCTGGAGGAGGATTGGAGCAGCCTGGAGGGTCATTGAAACTCTCATG
TGCAGCCTCTGGATTCACCTTCAATTCGTACGCCATGAACTGGGTCCGCCAGGCTCCAGGAAAGG
GTTTGGAATGGGTTGCTCGCATAAGAAGTAAATATAATAATTATGCAACATATTATGCCGATTCA
GTGAAAGGGAGGTTCACCATCTCCAGAGATGATTCAAAAAACACTGCCTATCTACAAATGAACAA
CTTGAAAACTGAGGACACTGCCGTGTACTACTGTGTGAGACATGGGAACTTCGGTAATAGCTACC
TATCCTTCTGGGCTTACTGGGGCCAAGGGACTCTGGTCACCGTCTCCTCAGGTGGTGGTGGTTCT
GGCGGCGGCGGCTCCGGTGGTGGTGGTTCTGAGCTCGTTGTGACTCAGGAACCTTCACTCACCGT
ATCACCTGGTGGAACAGTCACACTCACTTGTGGCTCCTCGACTGGGGCTGTTACATCTGGCTACT
ACCCAAACTGGGTCCAACAAAAACCAGGTCAGGCACCCCGTGGTCTAATAGGTGGGACTAAGTTC
CTCGCCCCCGGTACTCCTGCCAGATTCTCAGGCTCCCTGCTTGGAGGCAAGGCTGCCCTCACCCT
CTCAGGGGTACAGCCAGAGGATGAGGCAGAATATTACTGTGCTCTATGGTACAGCAACCGCTGGG
TGTTCGGTGGAGGAACCAAACTGACTGTCCTA 63. CDR-L1 of G4H artificial aa
GSSTGAVTSGYYPN 64. CDR-L2 of G4H artificial aa GTKFLAP 65. CDR-L3
of G4H artificial aa ALWYSNRWV 66. CDR-H1 of G4H artificial aa
RYAMN 67. CDR-H2 of G4H artificial aa RIRSKYNNYATYYADSVKG 68.
CDR-H3 of G4H artificial aa HGNFGNSYLSYFAY 69. VH of G4H artificial
aa
EVQLVESGGGLVQPGGSLKLSCAASGFTFNRYAMNWVRQAPGKGLEWVARIRSKYNNYATYYADS
VKGRFTISRDDSKNTAYLQMNNLKTEDTAVYYCVRHGNFGNSYLSYFAYWGQGTLVTVSS 70. VH
of G4H artificial nt
GAGGTGCAGCTGGTCGAGTCTGGAGGAGGATTGGTGCAGCCTGGAGGGTCATTGAAACTCTCATG
TGCAGCCTCTGGATTCACCTTCAATCGCTACGCCATGAACTGGGTCCGCCAGGCTCCAGGAAAGG
GTTTGGAATGGGTTGCTCGCATAAGAAGTAAATATAATAATTATGCAACATATTATGCCGATTCA
GTGAAAGGGAGGTTCACCATCTCCAGAGATGATTCAAAAAACACTGCCTATCTACAAATGAACAA
CTTGAAAACTGAGGACACTGCCGTGTACTACTGTGTGAGACATGGGAACTTCGGTAATAGCTACT
TATCCTACTTCGCTTACTGGGGCCAAGGGACTCTGGTCACCGTCTCCTCA 71. VL of G4H
artificial aa
QTVVTQEPSLTVSPGGTVTLTCGSSTGAVTSGYYPNWVQQKPGQAPRGLIGGTKFLAPGTPARFS
GSLLGGKAALTLSGVQPEDEAEYYCALWYSNRWVFGGGTKLTVL 72. VL of G4H
artificial nt
CAGACTGTTGTGACTCAGGAACCTTCACTCACCGTATCACCTGGTGGAACAGTCACACTCACTTG
TGGCTCCTCGACTGGGGCTGTTACATCTGGCTACTACCCAAACTGGGTCCAACAAAAACCAGGTC
AGGCACCCCGTGGTCTAATAGGTGGGACTAAGTTCCTCGCCCCCGGTACTCCTGCCAGATTCTCA
GGCTCCCTGCTTGGAGGCAAGGCTGCCCTCACCCTCTCAGGGGTACAGCCAGAGGATGAGGCAGA
ATATTACTGTGCTCTATGGTACAGCAACCGCTGGGTGTTCGGTGGAGGAACCAAACTGACTGTCC
TA 73. VH-P of G4H artificial aa
EVQLLESGGGLVQPGGSLKLSCAASGFTFNRYAMNWVRQAPGKGLEWVARIRSKYNNYATYYADS
VKGRFTISRDDSKNTAYLQMNNLKTEDTAVYYCVRHGNFGNSYLSYFAYWGQGTLVTVSS 74.
VH-P of G4H artificial nt
GAGGTGCAGCTGCTCGAGTCTGGAGGAGGATTGGTGCAGCCTGGAGGGTCATTGAAACTCTCATG
TGCAGCCTCTGGATTCACCTTCAATCGCTACGCCATGAACTGGGTCCGCCAGGCTCCAGGAAAGG
GTTTGGAATGGGTTGCTCGCATAAGAAGTAAATATAATAATTATGCAACATATTATGCCGATTCA
GTGAAAGGGAGGTTCACCATCTCCAGAGATGATTCAAAAAACACTGCCTATCTACAAATGAACAA
CTTGAAAACTGAGGACACTGCCGTGTACTACTGTGTGAGACATGGGAACTTCGGTAATAGCTACT
TATCCTACTTCGCTTACTGGGGCCAAGGGACTCTGGTCACCGTCTCCTCA 75. VL-P of G4H
artificial aa
ELVVTQEPSLTVSPGGTVTLTCGSSTGAVTSGYYPNWVQQKPGQAPRGLIGGTKFLAPGTPARFS
GSLLGGKAALTLSGVQPEDEAEYYCALWYSNRWVFGGGTKLTVL 76. VL-P of G4H
artificial nt
GAGCTCGTTGTGACTCAGGAACCTTCACTCACCGTATCACCTGGTGGAACAGTCACACTCACTTG
TGGCTCCTCGACTGGGGCTGTTACATCTGGCTACTACCCAAACTGGGTCCAACAAAAACCAGGTC
AGGCACCCCGTGGTCTAATAGGTGGGACTAAGTTCCTCGCCCCCGGTACTCCTGCCAGATTCTCA
GGCTCCCTGCTTGGAGGCAAGGCTGCCCTCACCCTCTCAGGGGTACAGCCAGAGGATGAGGCAGA
ATATTACTGTGCTCTATGGTACAGCAACCGCTGGGTGTTCGGTGGAGGAACCAAACTGACTGTCC
TA 77. VH-VL of G4H artificial aa
EVQLVESGGGLVQPGGSLKLSCAASGFTFNRYAMNWVRQAPGKGLEWVARIRSKYNNYATYYADS
VKGRFTISRDDSKNTAYLQMNNLKTEDTAVYYCVRHGNFGNSYLSYFAYWGQGTLVTVSSGGGGS
GGGGSGGGGSQTVVTQEPSLTVSPGGTVTLTCGSSTGAVTSGYYPNWVQQKPGQAPRGLIGGTKF
LAPGTPARFSGSLLGGKAALTLSGVQPEDEAEYYCALWYSNRWVFGGGTKLTVL 78. VH-VL of
G4H artificial nt
GAGGTGCAGCTGGTCGAGTCTGGAGGAGGATTGGTGCAGCCTGGAGGGTCATTGAAACTCTCATG
TGCAGCCTCTGGATTCACCTTCAATCGCTACGCCATGAACTGGGTCCGCCAGGCTCCAGGAAAGG
GTTTGGAATGGGTTGCTCGCATAAGAAGTAAATATAATAATTATGCAACATATTATGCCGATTCA
GTGAAAGGGAGGTTCACCATCTCCAGAGATGATTCAAAAAACACTGCCTATCTACAAATGAACAA
CTTGAAAACTGAGGACACTGCCGTGTACTACTGTGTGAGACATGGGAACTTCGGTAATAGCTACT
TATCCTACTTCGCTTACTGGGGCCAAGGGACTCTGGTCACCGTCTCCTCAGGTGGTGGTGGTTCT
GGCGGCGGCGGCTCCGGTGGTGGTGGTTCTCAGACTGTTGTGACTCAGGAACCTTCACTCACCGT
ATCACCTGGTGGAACAGTCACACTCACTTGTGGCTCCTCGACTGGGGCTGTTACATCTGGCTACT
ACCCAAACTGGGTCCAACAAAAACCAGGTCAGGCACCCCGTGGTCTAATAGGTGGGACTAAGTTC
CTCGCCCCCGGTACTCCTGCCAGATTCTCAGGCTCCCTGCTTGGAGGCAAGGCTGCCCTCACCCT
CTCAGGGGTACAGCCAGAGGATGAGGCAGAATATTACTGTGCTCTATGGTACAGCAACCGCTGGG
TGTTCGGTGGAGGAACCAAACTGACTGTCCTA 79. VH-VL-P of G4H artificial aa
EVQLLESGGGLVQPGGSLKLSCAASGFTFNRYAMNWVRQAPGKGLEWVARIRSKYNNYATYYADS
VKGRFTISRDDSKNTAYLQMNNLKTEDTAVYYCVRHGNFGNSYLSYFAYWGQGTLVTVSSGGGGS
GGGGSGGGGSELVVTQEPSLTVSPGGTVTLTCGSSTGAVTSGYYPNWVQQKPGQAPRGLIGGTKF
LAPGTPARFSGSLLGGKAALTLSGVQPEDEAEYYCALWYSNRWVFGGGTKLTVL 80. VH-VL-P
of G4H artificial nt
GAGGTGCAGCTGCTCGAGTCTGGAGGAGGATTGGTGCAGCCTGGAGGGTCATTGAAACTCTCATG
TGCAGCCTCTGGATTCACCTTCAATCGCTACGCCATGAACTGGGTCCGCCAGGCTCCAGGAAAGG
GTTTGGAATGGGTTGCTCGCATAAGAAGTAAATATAATAATTATGCAACATATTATGCCGATTCA
GTGAAAGGGAGGTTCACCATCTCCAGAGATGATTCAAAAAACACTGCCTATCTACAAATGAACAA
CTTGAAAACTGAGGACACTGCCGTGTACTACTGTGTGAGACATGGGAACTTCGGTAATAGCTACT
TATCCTACTTCGCTTACTGGGGCCAAGGGACTCTGGTCACCGTCTCCTCAGGTGGTGGTGGTTCT
GGCGGCGGCGGCTCCGGTGGTGGTGGTTCTGAGCTCGTTGTGACTCAGGAACCTTCACTCACCGT
ATCACCTGGTGGAACAGTCACACTCACTTGTGGCTCCTCGACTGGGGCTGTTACATCTGGCTACT
ACCCAAACTGGGTCCAACAAAAACCAGGTCAGGCACCCCGTGGTCTAATAGGTGGGACTAAGTTC
CTCGCCCCCGGTACTCCTGCCAGATTCTCAGGCTCCCTGCTTGGAGGCAAGGCTGCCCTCACCCT
CTCAGGGGTACAGCCAGAGGATGAGGCAGAATATTACTGTGCTCTATGGTACAGCAACCGCTGGG
TGTTCGGTGGAGGAACCAAACTGACTGTCCTA 81. CDR-L1 of A2J artificial aa
RSSTGAVTSGYYPN 82. CDR-L2 of A2J artificial aa ATDMRPS 83. CDR-L3
of A2J artificial aa ALWYSNRWV 84. CDR-H1 of A2J artificial aa
VYAMN 85. CDR-H2 of A2J artificial aa RIRSKYNNYATYYADSVKK 86.
CDR-H3 of A2J artificial aa HGNFGNSYLSWWAY 87. VH of A2J artificial
aa
EVQLVESGGGLVQPGGSLKLSCAASGFTFNVYAMNWVRQAPGKGLEWVARIRSKYNNYATYYADS
VKKRFTISRDDSKNTAYLQMNNLKTEDTAVYYCVRHGNFGNSYLSWWAYWGQGTLVTVSS 88. VH
of A2J artificial nt
GAGGTGCAGCTGGTCGAGTCTGGAGGAGGATTGGTGCAGCCTGGAGGGTCATTGAAACTCTCATG
TGCAGCCTCTGGATTCACCTTCAATGTCTACGCCATGAACTGGGTCCGCCAGGCTCCAGGAAAGG
GTTTGGAATGGGTTGCTCGCATAAGAAGTAAATATAATAATTATGCAACATATTATGCCGATTCA
GTGAAAAAGAGGTTCACCATCTCCAGAGATGATTCAAAAAACACTGCCTATCTACAAATGAACAA
CTTGAAAACTGAGGACACTGCCGTGTACTACTGTGTGAGACATGGGAACTTCGGTAATAGCTACT
TATCCTGGTGGGCTTACTGGGGCCAAGGGACTCTGGTCACCGTCTCCTCA 89. VL of A2J
artificial aa
QTVVTQEPSLTVSPGGTVTLTCRSSTGAVTSGYYPNWVQQKPGQAPRGLIGATDMRPSGTPARFS
GSLLGGKAALTLSGVQPEDEAEYYCALWYSNRWVFGGGTKLTVL 90. VL of A2J
artificial nt
CAGACTGTTGTGACTCAGGAACCTTCACTCACCGTATCACCTGGTGGAACAGTCACACTCACTTG
TCGCTCCTCGACTGGGGCTGTTACATCTGGCTACTACCCAAACTGGGTCCAACAAAAACCAGGTC
AGGCACCCCGTGGTCTAATAGGTGCCACTGACATGAGGCCCTCTGGTACTCCTGCCAGATTCTCA
GGCTCCCTGCTTGGAGGCAAGGCTGCCCTCACCCTCTCAGGGGTACAGCCAGAGGATGAGGCAGA
ATATTACTGTGCTCTATGGTACAGCAACCGCTGGGTGTTCGGTGGAGGAACCAAACTGACTGTCC
TA 91. VH-P of A2J artificial aa
EVQLLESGGGLVQPGGSLKLSCAASGFTFNVYAMNWVRQAPGKGLEWVARIRSKYNNYATYYADS
VKKRFTISRDDSKNTAYLQMNNLKTEDTAVYYCVRHGNFGNSYLSWWAYWGQGTLVTVSS 92.
VH-P of A2J artificial nt
GAGGTGCAGCTGCTCGAGTCTGGAGGAGGATTGGTGCAGCCTGGAGGGTCATTGAAACTCTCATG
TGCAGCCTCTGGATTCACCTTCAATGTCTACGCCATGAACTGGGTCCGCCAGGCTCCAGGAAAGG
GTTTGGAATGGGTTGCTCGCATAAGAAGTAAATATAATAATTATGCAACATATTATGCCGATTCA
GTGAAAAAGAGGTTCACCATCTCCAGAGATGATTCAAAAAACACTGCCTATCTACAAATGAACAA
CTTGAAAACTGAGGACACTGCCGTGTACTACTGTGTGAGACATGGGAACTTCGGTAATAGCTACT
TATCCTGGTGGGCTTACTGGGGCCAAGGGACTCTGGTCACCGTCTCCTCA 93. VL-P of A2J
artificial aa
ELVVTQEPSLTVSPGGTVTLTCRSSTGAVTSGYYPNWVQQKPGQAPRGLIGATDMRPSGTPARFS
GSLLGGKAALTLSGVQPEDEAEYYCALWYSNRWVFGGGTKLTVL 94. VL-P of A2J
artificial nt
GAGCTCGTTGTGACTCAGGAACCTTCACTCACCGTATCACCTGGTGGAACAGTCACACTCACTTG
TCGCTCCTCGACTGGGGCTGTTACATCTGGCTACTACCCAAACTGGGTCCAACAAAAACCAGGTC
AGGCACCCCGTGGTCTAATAGGTGCCACTGACATGAGGCCCTCTGGTACTCCTGCCAGATTCTCA
GGCTCCCTGCTTGGAGGCAAGGCTGCCCTCACCCTCTCAGGGGTACAGCCAGAGGATGAGGCAGA
ATATTACTGTGCTCTATGGTACAGCAACCGCTGGGTGTTCGGTGGAGGAACCAAACTGACTGTCC
TA 95. VH-VL of A2J artificial aa
EVQLVESGGGLVQPGGSLKLSCAASGFTFNVYAMNWVRQAPGKGLEWVARIRSKYNNYATYYADS
VKKRFTISRDDSKNTAYLQMNNLKTEDTAVYYCVRHGNFGNSYLSWWAYWGQGTLVTVSSGGGGS
GGGGSGGGGSQTVVTQEPSLTVSPGGTVTLTCRSSTGAVTSGYYPNWVQQKPGQAPRGLIGATDM
RPSGTPARFSGSLLGGKAALTLSGVQPEDEAEYYCALWYSNRWVFGGGTKLTVL 96. VH-VL of
A2J artificial nt
GAGGTGCAGCTGGTCGAGTCTGGAGGAGGATTGGTGCAGCCTGGAGGGTCATTGAAACTCTCATG
TGCAGCCTCTGGATTCACCTTCAATGTCTACGCCATGAACTGGGTCCGCCAGGCTCCAGGAAAGG
GTTTGGAATGGGTTGCTCGCATAAGAAGTAAATATAATAATTATGCAACATATTATGCCGATTCA
GTGAAAAAGAGGTTCACCATCTCCAGAGATGATTCAAAAAACACTGCCTATCTACAAATGAACAA
CTTGAAAACTGAGGACACTGCCGTGTACTACTGTGTGAGACATGGGAACTTCGGTAATAGCTACT
TATCCTGGTGGGCTTACTGGGGCCAAGGGACTCTGGTCACCGTCTCCTCAGGTGGTGGTGGTTCT
GGCGGCGGCGGCTCCGGTGGTGGTGGTTCTCAGACTGTTGTGACTCAGGAACCTTCACTCACCGT
ATCACCTGGTGGAACAGTCACACTCACTTGTCGCTCCTCGACTGGGGCTGTTACATCTGGCTACT
ACCCAAACTGGGTCCAACAAAAACCAGGTCAGGCACCCCGTGGTCTAATAGGTGCCACTGACATG
AGGCCCTCTGGTACTCCTGCCAGATTCTCAGGCTCCCTGCTTGGAGGCAAGGCTGCCCTCACCCT
CTCAGGGGTACAGCCAGAGGATGAGGCAGAATATTACTGTGCTCTATGGTACAGCAACCGCTGGG
TGTTCGGTGGAGGAACCAAACTGACTGTCCTA 97. VH-VL-P of A2J artificial aa
EVQLLESGGGLVQPGGSLKLSCAASGFTFNVYAMNWVRQAPGKGLEWVARIRSKYNNYATYYADS
VKKRFTISRDDSKNTAYLQMNNLKTEDTAVYYCVRHGNFGNSYLSWWAYWGQGTLVTVSSGGGGS
GGGGSGGGGSELVVTQEPSLTVSPGGTVTLTCRSSTGAVTSGYYPNWVQQKPGQAPRGLIGATDM
RPSGTPARFSGSLLGGKAALTLSGVQPEDEAEYYCALWYSNRWVFGGGTKLTVL 98. VH-VL-P
of A2J artificial nt
GAGGTGCAGCTGCTCGAGTCTGGAGGAGGATTGGTGCAGCCTGGAGGGTCATTGAAACTCTCATG
TGCAGCCTCTGGATTCACCTTCAATGTCTACGCCATGAACTGGGTCCGCCAGGCTCCAGGAAAGG
GTTTGGAATGGGTTGCTCGCATAAGAAGTAAATATAATAATTATGCAACATATTATGCCGATTCA
GTGAAAAAGAGGTTCACCATCTCCAGAGATGATTCAAAAAACACTGCCTATCTACAAATGAACAA
CTTGAAAACTGAGGACACTGCCGTGTACTACTGTGTGAGACATGGGAACTTCGGTAATAGCTACT
TATCCTGGTGGGCTTACTGGGGCCAAGGGACTCTGGTCACCGTCTCCTCAGGTGGTGGTGGTTCT
GGCGGCGGCGGCTCCGGTGGTGGTGGTTCTGAGCTCGTTGTGACTCAGGAACCTTCACTCACCGT
ATCACCTGGTGGAACAGTCACACTCACTTGTCGCTCCTCGACTGGGGCTGTTACATCTGGCTACT
ACCCAAACTGGGTCCAACAAAAACCAGGTCAGGCACCCCGTGGTCTAATAGGTGCCACTGACATG
AGGCCCTCTGGTACTCCTGCCAGATTCTCAGGCTCCCTGCTTGGAGGCAAGGCTGCCCTCACCCT
CTCAGGGGTACAGCCAGAGGATGAGGCAGAATATTACTGTGCTCTATGGTACAGCAACCGCTGGG
TGTTCGGTGGAGGAACCAAACTGACTGTCCTA 99. CDR-L1 of E1L artificial aa
GSSTGAVTSGYYPN 100. CDR-L2 of E1L artificial aa GTKFLAP 101. CDR-L3
of E1L artificial aa ALWYSNRWV 102. CDR-H1 of E1L artificial aa
KYAMN 103. CDR-H2 of E1L artificial aa RIRSKYNNYATYYADSVKS 104.
CDR-H3 of E1L artificial aa HGNFGNSYTSYYAY 105. VH of E1L
artificial aa
EVQLVESGGGLVQPGGSLKLSCAASGFTFNKYAMNWVRQAPGKGLEWVARIRSKYNNYATYYADS
VKSRFTISRDDSKNTAYLQMNNLKTEDTAVYYCVRHGNFGNSYTSYYAYWGQGTLVTVSS 106.
VH of E1L artificial nt
GAGGTGCAGCTGGTCGAGTCTGGAGGAGGATTGGTGCAGCCTGGAGGGTCATTGAAACTCTCATG
TGCAGCCTCTGGATTCACCTTCAATAAGTACGCCATGAACTGGGTCCGCCAGGCTCCAGGAAAGG
GTTTGGAATGGGTTGCTCGCATAAGAAGTAAATATAATAATTATGCAACATATTATGCCGATTCA
GTGAAATCGAGGTTCACCATCTCCAGAGATGATTCAAAAAACACTGCCTATCTACAAATGAACAA
CTTGAAAACTGAGGACACTGCCGTGTACTACTGTGTGAGACATGGGAACTTCGGTAATAGCTACA
CATCCTACTACGCTTACTGGGGCCAAGGGACTCTGGTCACCGTCTCCTCA 107. VL of E1L
artificial aa
QTVVTQEPSLTVSPGGTVTLTCGSSTGAVTSGYYPNWVQQKPGQAPRGLIGGTKFLAPGTPARFS
GSLLGGKAALTLSGVQPEDEAEYYCALWYSNRWVFGGGTKLTVL 108. VL of E1L
artificial nt
CAGACTGTTGTGACTCAGGAACCTTCACTCACCGTATCACCTGGTGGAACAGTCACACTCACTTG
TGGCTCCTCGACTGGGGCTGTTACATCTGGCTACTACCCAAACTGGGTCCAACAAAAACCAGGTC
AGGCACCCCGTGGTCTAATAGGTGGGACTAAGTTCCTCGCCCCCGGTACTCCTGCCAGATTCTCA
GGCTCCCTGCTTGGAGGCAAGGCTGCCCTCACCCTCTCAGGGGTACAGCCAGAGGATGAGGCAGA
ATATTACTGTGCTCTATGGTACAGCAACCGCTGGGTGTTCGGTGGAGGAACCAAACTGACTGTCC
TA 109. VH-P of E1L artificial aa
EVQLLESGGGLVQPGGSLKLSCAASGFTFNKYAMNWVRQAPGKGLEWVARIRSKYNNYATYYADS
VKSRFTISRDDSKNTAYLQMNNLKTEDTAVYYCVRHGNFGNSYTSYYAYWGQGTLVTVSS 110.
VH-P of E1L artificial nt
GAGGTGCAGCTGCTCGAGTCTGGAGGAGGATTGGTGCAGCCTGGAGGGTCATTGAAACTCTCATG
TGCAGCCTCTGGATTCACCTTCAATAAGTACGCCATGAACTGGGTCCGCCAGGCTCCAGGAAAGG
GTTTGGAATGGGTTGCTCGCATAAGAAGTAAATATAATAATTATGCAACATATTATGCCGATTCA
GTGAAATCGAGGTTCACCATCTCCAGAGATGATTCAAAAAACACTGCCTATCTACAAATGAACAA
CTTGAAAACTGAGGACACTGCCGTGTACTACTGTGTGAGACATGGGAACTTCGGTAATAGCTACA
CATCCTACTACGCTTACTGGGGCCAAGGGACTCTGGTCACCGTCTCCTCA 111. VL-P of E1L
artificial aa
ELVVTQEPSLTVSPGGTVTLTCGSSTGAVTSGYYPNWVQQKPGQAPRGLIGGTKFLAPGTPARFS
GSLLGGKAALTLSGVQPEDEAEYYCALWYSNRWVFGGGTKLTVL 112. VL-P of E1L
artificial nt
GAGCTCGTTGTGACTCAGGAACCTTCACTCACCGTATCACCTGGTGGAACAGTCACACTCACTTG
TGGCTCCTCGACTGGGGCTGTTACATCTGGCTACTACCCAAACTGGGTCCAACAAAAACCAGGTC
AGGCACCCCGTGGTCTAATAGGTGGGACTAAGTTCCTCGCCCCCGGTACTCCTGCCAGATTCTCA
GGCTCCCTGCTTGGAGGCAAGGCTGCCCTCACCCTCTCAGGGGTACAGCCAGAGGATGAGGCAGA
ATATTACTGTGCTCTATGGTACAGCAACCGCTGGGTGTTCGGTGGAGGAACCAAACTGACTGTCC
TA 113. VH-VL of E1L artificial aa
EVQLVESGGGLVQPGGSLKLSCAASGFTFNKYAMNWVRQAPGKGLEWVARIRSKYNNYATYYADS
VKSRFTISRDDSKNTAYLQMNNLKTEDTAVYYCVRHGNFGNSYTSYYAYWGQGTLVTVSSGGGGS
GGGGSGGGGSQTVVTQEPSLTVSPGGTVTLTCGSSTGAVTSGYYPNWVQQKPGQAPRGLIGGTKF
LAPGTPARFSGSLLGGKAALTLSGVQPEDEAEYYCALWYSNRWVFGGGTKLTVL 114. VH-VL
of E1L artificial nt
GAGGTGCAGCTGGTCGAGTCTGGAGGAGGATTGGTGCAGCCTGGAGGGTCATTGAAACTCTCATG
TGCAGCCTCTGGATTCACCTTCAATAAGTACGCCATGAACTGGGTCCGCCAGGCTCCAGGAAAGG
GTTTGGAATGGGTTGCTCGCATAAGAAGTAAATATAATAATTATGCAACATATTATGCCGATTCA
GTGAAATCGAGGTTCACCATCTCCAGAGATGATTCAAAAAACACTGCCTATCTACAAATGAACAA
CTTGAAAACTGAGGACACTGCCGTGTACTACTGTGTGAGACATGGGAACTTCGGTAATAGCTACA
CATCCTACTACGCTTACTGGGGCCAAGGGACTCTGGTCACCGTCTCCTCAGGTGGTGGTGGTTCT
GGCGGCGGCGGCTCCGGTGGTGGTGGTTCTCAGACTGTTGTGACTCAGGAACCTTCACTCACCGT
ATCACCTGGTGGAACAGTCACACTCACTTGTGGCTCCTCGACTGGGGCTGTTACATCTGGCTACT
ACCCAAACTGGGTCCAACAAAAACCAGGTCAGGCACCCCGTGGTCTAATAGGTGGGACTAAGTTC
CTCGCCCCCGGTACTCCTGCCAGATTCTCAGGCTCCCTGCTTGGAGGCAAGGCTGCCCTCACCCT
CTCAGGGGTACAGCCAGAGGATGAGGCAGAATATTACTGTGCTCTATGGTACAGCAACCGCTGGG
TGTTCGGTGGAGGAACCAAACTGACTGTCCTA 115. VH-VL-P of E1L artificial aa
EVQLLESGGGLVQPGGSLKLSCAASGFTFNKYAMNWVRQAPGKGLEWVARIRSKYNNYATYYADS
VKSRFTISRDDSKNTAYLQMNNLKTEDTAVYYCVRHGNFGNSYTSYYAYWGQGTLVTVSSGGGGS
GGGGSGGGGSELVVTQEPSLTVSPGGTVTLTCGSSTGAVTSGYYPNWVQQKPGQAPRGLIGGTKF
LAPGTPARFSGSLLGGKAALTLSGVQPEDEAEYYCALWYSNRWVFGGGTKLTVL 116. VH-VL-P
of E1L artificial nt
GAGGTGCAGCTGCTCGAGTCTGGAGGAGGATTGGTGCAGCCTGGAGGGTCATTGAAACTCTCATG
TGCAGCCTCTGGATTCACCTTCAATAAGTACGCCATGAACTGGGTCCGCCAGGCTCCAGGAAAGG
GTTTGGAATGGGTTGCTCGCATAAGAAGTAAATATAATAATTATGCAACATATTATGCCGATTCA
GTGAAATCGAGGTTCACCATCTCCAGAGATGATTCAAAAAACACTGCCTATCTACAAATGAACAA
CTTGAAAACTGAGGACACTGCCGTGTACTACTGTGTGAGACATGGGAACTTCGGTAATAGCTACA
CATCCTACTACGCTTACTGGGGCCAAGGGACTCTGGTCACCGTCTCCTCAGGTGGTGGTGGTTCT
GGCGGCGGCGGCTCCGGTGGTGGTGGTTCTGAGCTCGTTGTGACTCAGGAACCTTCACTCACCGT
ATCACCTGGTGGAACAGTCACACTCACTTGTGGCTCCTCGACTGGGGCTGTTACATCTGGCTACT
ACCCAAACTGGGTCCAACAAAAACCAGGTCAGGCACCCCGTGGTCTAATAGGTGGGACTAAGTTC
CTCGCCCCCGGTACTCCTGCCAGATTCTCAGGCTCCCTGCTTGGAGGCAAGGCTGCCCTCACCCT
CTCAGGGGTACAGCCAGAGGATGAGGCAGAATATTACTGTGCTCTATGGTACAGCAACCGCTGGG
TGTTCGGTGGAGGAACCAAACTGACTGTCCTA 117. CDR-L1 of E2M artificial aa
RSSTGAVTSGYYPN 118. CDR-L2 of E2M artificial aa ATDMRPS 119. CDR-L3
of E2M artificial aa ALWYSNRWV 120. CDR-H1 of E2M artificial aa
GYAMN 121. CDR-H2 of E2M artificial aa RIRSKYNNYATYYADSVKE 122.
CDR-H3 of E2M artificial aa HRNFGNSYLSWFAY 123. VH of E2M
artificial aa
EVQLVESGGGLVQPGGSLKLSCAASGFTFNGYAMNWVRQAPGKGLEWVARIRSKYNNYATYYADS
VKERFTISRDDSKNTAYLQMNNLKTEDTAVYYCVRHRNFGNSYLSWFAYWGQGTLVTVSS 124.
VH of E2M artificial nt
GAGGTGCAGCTGGTCGAGTCTGGAGGAGGATTGGTGCAGCCTGGAGGGTCATTGAAACTCTCATG
TGCAGCCTCTGGATTCACCTTCAATGGCTACGCCATGAACTGGGTCCGCCAGGCTCCAGGAAAGG
GTTTGGAATGGGTTGCTCGCATAAGAAGTAAATATAATAATTATGCAACATATTATGCCGATTCA
GTGAAAGAGAGGTTCACCATCTCCAGAGATGATTCAAAAAACACTGCCTATCTACAAATGAACAA
CTTGAAAACTGAGGACACTGCCGTGTACTACTGTGTGAGACATAGGAACTTCGGTAATAGCTACT
TATCCTGGTTCGCTTACTGGGGCCAAGGGACTCTGGTCACCGTCTCCTCA 125. VL of E2M
artificial aa
QTVVTQEPSLTVSPGGTVTLTCRSSTGAVTSGYYPNWVQQKPGQAPRGLIGATDMRPSGTPARFS
GSLLGGKAALTLSGVQPEDEAEYYCALWYSNRWVFGGGTKLTVL 126. VL of E2M
artificial nt
CAGACTGTTGTGACTCAGGAACCTTCACTCACCGTATCACCTGGTGGAACAGTCACACTCACTTG
TCGCTCCTCGACTGGGGCTGTTACATCTGGCTACTACCCAAACTGGGTCCAACAAAAACCAGGTC
AGGCACCCCGTGGTCTAATAGGTGCCACTGACATGAGGCCCTCTGGTACTCCTGCCAGATTCTCA
GGCTCCCTGCTTGGAGGCAAGGCTGCCCTCACCCTCTCAGGGGTACAGCCAGAGGATGAGGCAGA
ATATTACTGTGCTCTATGGTACAGCAACCGCTGGGTGTTCGGTGGAGGAACCAAACTGACTGTCC
TA 127. VH-P of E2M artificial aa
EVQLLESGGGLVQPGGSLKLSCAASGFTFNGYAMNWVRQAPGKGLEWVARIRSKYNNYATYYADS
VKERFTISRDDSKNTAYLQMNNLKTEDTAVYYCVRHRNFGNSYLSWFAYWGQGTLVTVSS 128.
VH-P of E2M artificial nt
GAGGTGCAGCTGCTCGAGTCTGGAGGAGGATTGGTGCAGCCTGGAGGGTCATTGAAACTCTCATG
TGCAGCCTCTGGATTCACCTTCAATGGCTACGCCATGAACTGGGTCCGCCAGGCTCCAGGAAAGG
GTTTGGAATGGGTTGCTCGCATAAGAAGTAAATATAATAATTATGCAACATATTATGCCGATTCA
GTGAAAGAGAGGTTCACCATCTCCAGAGATGATTCAAAAAACACTGCCTATCTACAAATGAACAA
CTTGAAAACTGAGGACACTGCCGTGTACTACTGTGTGAGACATAGGAACTTCGGTAATAGCTACT
TATCCTGGTTCGCTTACTGGGGCCAAGGGACTCTGGTCACCGTCTCCTCA 129. VL-P of E2M
artificial aa
ELVVTQEPSLTVSPGGTVTLTCRSSTGAVTSGYYPNWVQQKPGQAPRGLIGATDMRPSGTPARFS
GSLLGGKAALTLSGVQPEDEAEYYCALWYSNRWVFGGGTKLTVL 130. VL-P of E2M
artificial nt
GAGCTCGTTGTGACTCAGGAACCTTCACTCACCGTATCACCTGGTGGAACAGTCACACTCACTTG
TCGCTCCTCGACTGGGGCTGTTACATCTGGCTACTACCCAAACTGGGTCCAACAAAAACCAGGTC
AGGCACCCCGTGGTCTAATAGGTGCCACTGACATGAGGCCCTCTGGTACTCCTGCCAGATTCTCA
GGCTCCCTGCTTGGAGGCAAGGCTGCCCTCACCCTCTCAGGGGTACAGCCAGAGGATGAGGCAGA
ATATTACTGTGCTCTATGGTACAGCAACCGCTGGGTGTTCGGTGGAGGAACCAAACTGACTGTCC
TA 131. VH-VL of E2M artificial aa
EVQLVESGGGLVQPGGSLKLSCAASGFTFNGYAMNWVRQAPGKGLEWVARIRSKYNNYATYYADS
VKERFTISRDDSKNTAYLQMNNLKTEDTAVYYCVRHRNFGNSYLSWFAYWGQGTLVTVSSGGGGS
GGGGSGGGGSQTVVTQEPSLTVSPGGTVTLTCRSSTGAVTSGYYPNWVQQKPGQAPRGLIGATDM
RPSGTPARFSGSLLGGKAALTLSGVQPEDEAEYYCALWYSNRWVFGGGTKLTVL 132. VH-VL
of E2M artificial nt
GAGGTGCAGCTGGTCGAGTCTGGAGGAGGATTGGTGCAGCCTGGAGGGTCATTGAAACTCTCATG
TGCAGCCTCTGGATTCACCTTCAATGGCTACGCCATGAACTGGGTCCGCCAGGCTCCAGGAAAGG
GTTTGGAATGGGTTGCTCGCATAAGAAGTAAATATAATAATTATGCAACATATTATGCCGATTCA
GTGAAAGAGAGGTTCACCATCTCCAGAGATGATTCAAAAAACACTGCCTATCTACAAATGAACAA
CTTGAAAACTGAGGACACTGCCGTGTACTACTGTGTGAGACATAGGAACTTCGGTAATAGCTACT
TATCCTGGTTCGCTTACTGGGGCCAAGGGACTCTGGTCACCGTCTCCTCAGGTGGTGGTGGTTCT
GGCGGCGGCGGCTCCGGTGGTGGTGGTTCTCAGACTGTTGTGACTCAGGAACCTTCACTCACCGT
ATCACCTGGTGGAACAGTCACACTCACTTGTCGCTCCTCGACTGGGGCTGTTACATCTGGCTACT
ACCCAAACTGGGTCCAACAAAAACCAGGTCAGGCACCCCGTGGTCTAATAGGTGCCACTGACATG
AGGCCCTCTGGTACTCCTGCCAGATTCTCAGGCTCCCTGCTTGGAGGCAAGGCTGCCCTCACCCT
CTCAGGGGTACAGCCAGAGGATGAGGCAGAATATTACTGTGCTCTATGGTACAGCAACCGCTGGG
TGTTCGGTGGAGGAACCAAACTGACTGTCCTA 133. VH-VL-P of E2M artificial aa
EVQLLESGGGLVQPGGSLKLSCAASGFTFNGYAMNWVRQAPGKGLEWVARIRSKYNNYATYYADS
VKERFTISRDDSKNTAYLQMNNLKTEDTAVYYCVRHRNFGNSYLSWFAYWGQGTLVTVSSGGGGS
GGGGSGGGGSELVVTQEPSLTVSPGGTVTLTCRSSTGAVTSGYYPNWVQQKPGQAPRGLIGATDM
RPSGTPARFSGSLLGGKAALTLSGVQPEDEAEYYCALWYSNRWVFGGGTKLTVL 134. VH-VL-P
of E2M artificial nt
GAGGTGCAGCTGCTCGAGTCTGGAGGAGGATTGGTGCAGCCTGGAGGGTCATTGAAACTCTCATG
TGCAGCCTCTGGATTCACCTTCAATGGCTACGCCATGAACTGGGTCCGCCAGGCTCCAGGAAAGG
GTTTGGAATGGGTTGCTCGCATAAGAAGTAAATATAATAATTATGCAACATATTATGCCGATTCA
GTGAAAGAGAGGTTCACCATCTCCAGAGATGATTCAAAAAACACTGCCTATCTACAAATGAACAA
CTTGAAAACTGAGGACACTGCCGTGTACTACTGTGTGAGACATAGGAACTTCGGTAATAGCTACT
TATCCTGGTTCGCTTACTGGGGCCAAGGGACTCTGGTCACCGTCTCCTCAGGTGGTGGTGGTTCT
GGCGGCGGCGGCTCCGGTGGTGGTGGTTCTGAGCTCGTTGTGACTCAGGAACCTTCACTCACCGT
ATCACCTGGTGGAACAGTCACACTCACTTGTCGCTCCTCGACTGGGGCTGTTACATCTGGCTACT
ACCCAAACTGGGTCCAACAAAAACCAGGTCAGGCACCCCGTGGTCTAATAGGTGCCACTGACATG
AGGCCCTCTGGTACTCCTGCCAGATTCTCAGGCTCCCTGCTTGGAGGCAAGGCTGCCCTCACCCT
CTCAGGGGTACAGCCAGAGGATGAGGCAGAATATTACTGTGCTCTATGGTACAGCAACCGCTGGG
TGTTCGGTGGAGGAACCAAACTGACTGTCCTA 135. CDR-L1 of F7O artificial aa
GSSTGAVTSGYYPN 136. CDR-L2 of F7O artificial aa GTKFLAP 137. CDR-L3
of F7O artificial aa ALWYSNRWV 138. CDR-H1 of F7O artificial aa
VYAMN 139. CDR-H2 of F7O artificial aa RIRSKYNNYATYYADSVKK 140.
CDR-H3 of F7O artificial aa HGNFGNSYISWWAY 141. VH of F7O
artificial aa
EVQLVESGGGLVQPGGSLKLSCAASGFTFNVYAMNWVRQAPGKGLEWVARIRSKYNNYATYYADS
VKKRFTISRDDSKNTAYLQMNNLKTEDTAVYYCVRHGNFGNSYISWWAYWGQGTLVTVSS 142.
VH of F7O artificial nt
GAGGTGCAGCTGGTCGAGTCTGGAGGAGGATTGGTGCAGCCTGGAGGGTCATTGAAACTCTCATG
TGCAGCCTCTGGATTCACCTTCAATGTGTACGCCATGAACTGGGTCCGCCAGGCTCCAGGAAAGG
GTTTGGAATGGGTTGCTCGCATAAGAAGTAAATATAATAATTATGCAACATATTATGCCGATTCA
GTGAAAAAGAGGTTCACCATCTCCAGAGATGATTCAAAAAACACTGCCTATCTACAAATGAACAA
CTTGAAAACTGAGGACACTGCCGTGTACTACTGTGTGAGACATGGGAACTTCGGTAATAGCTACA
TATCCTGGTGGGCTTACTGGGGCCAAGGGACTCTGGTCACCGTCTCCTCA 143. VL of F7O
artificial aa
QTVVTQEPSLTVSPGGTVTLTCGSSTGAVTSGYYPNWVQQKPGQAPRGLIGGTKFLAPGTPARFS
GSLLGGKAALTLSGVQPEDEAEYYCALWYSNRWVFGGGTKLTVL 144. VL of F7O
artificial nt
CAGACTGTTGTGACTCAGGAACCTTCACTCACCGTATCACCTGGTGGAACAGTCACACTCACTTG
TGGCTCCTCGACTGGGGCTGTTACATCTGGCTACTACCCAAACTGGGTCCAACAAAAACCAGGTC
AGGCACCCCGTGGTCTAATAGGTGGGACTAAGTTCCTCGCCCCCGGTACTCCTGCCAGATTCTCA
GGCTCCCTGCTTGGAGGCAAGGCTGCCCTCACCCTCTCAGGGGTACAGCCAGAGGATGAGGCAGA
ATATTACTGTGCTCTATGGTACAGCAACCGCTGGGTGTTCGGTGGAGGAACCAAACTGACTGTCC
TA 145. VH-P of F7O artificial aa
EVQLLESGGGLVQPGGSLKLSCAASGFTFNVYAMNWVRQAPGKGLEWVARIRSKYNNYATYYADS
VKKRFTISRDDSKNTAYLQMNNLKTEDTAVYYCVRHGNFGNSYISWWAYWGQGTLVTVSS 146.
VH-P of F7O artificial nt
GAGGTGCAGCTGCTCGAGTCTGGAGGAGGATTGGTGCAGCCTGGAGGGTCATTGAAACTCTCATG
TGCAGCCTCTGGATTCACCTTCAATGTGTACGCCATGAACTGGGTCCGCCAGGCTCCAGGAAAGG
GTTTGGAATGGGTTGCTCGCATAAGAAGTAAATATAATAATTATGCAACATATTATGCCGATTCA
GTGAAAAAGAGGTTCACCATCTCCAGAGATGATTCAAAAAACACTGCCTATCTACAAATGAACAA
CTTGAAAACTGAGGACACTGCCGTGTACTACTGTGTGAGACATGGGAACTTCGGTAATAGCTACA
TATCCTGGTGGGCTTACTGGGGCCAAGGGACTCTGGTCACCGTCTCCTCA 147. VL-P of F7O
artificial aa
ELVVTQEPSLTVSPGGTVTLTCGSSTGAVTSGYYPNWVQQKPGQAPRGLIGGTKFLAPGTPARFS
GSLLGGKAALTLSGVQPEDEAEYYCALWYSNRWVFGGGTKLTVL 148. VL-P of F7O
artificial nt
GAGCTCGTTGTGACTCAGGAACCTTCACTCACCGTATCACCTGGTGGAACAGTCACACTCACTTG
TGGCTCCTCGACTGGGGCTGTTACATCTGGCTACTACCCAAACTGGGTCCAACAAAAACCAGGTC
AGGCACCCCGTGGTCTAATAGGTGGGACTAAGTTCCTCGCCCCCGGTACTCCTGCCAGATTCTCA
GGCTCCCTGCTTGGAGGCAAGGCTGCCCTCACCCTCTCAGGGGTACAGCCAGAGGATGAGGCAGA
ATATTACTGTGCTCTATGGTACAGCAACCGCTGGGTGTTCGGTGGAGGAACCAAACTGACTGTCC
TA 149. VH-VL of F7O artificial aa
EVQLVESGGGLVQPGGSLKLSCAASGFTFNVYAMNWVRQAPGKGLEWVARIRSKYNNYATYYADS
VKKRFTISRDDSKNTAYLQMNNLKTEDTAVYYCVRHGNFGNSYISWWAYWGQGTLVTVSSGGGGS
GGGGSGGGGSQTVVTQEPSLTVSPGGTVTLTCGSSTGAVTSGYYPNWVQQKPGQAPRGLIGGTKF
LAPGTPARFSGSLLGGKAALTLSGVQPEDEAEYYCALWYSNRWVFGGGTKLTVL 150. VH-VL
of F7O artificial nt
GAGGTGCAGCTGGTCGAGTCTGGAGGAGGATTGGTGCAGCCTGGAGGGTCATTGAAACTCTCATG
TGCAGCCTCTGGATTCACCTTCAATGTGTACGCCATGAACTGGGTCCGCCAGGCTCCAGGAAAGG
GTTTGGAATGGGTTGCTCGCATAAGAAGTAAATATAATAATTATGCAACATATTATGCCGATTCA
GTGAAAAAGAGGTTCACCATCTCCAGAGATGATTCAAAAAACACTGCCTATCTACAAATGAACAA
CTTGAAAACTGAGGACACTGCCGTGTACTACTGTGTGAGACATGGGAACTTCGGTAATAGCTACA
TATCCTGGTGGGCTTACTGGGGCCAAGGGACTCTGGTCACCGTCTCCTCAGGTGGTGGTGGTTCT
GGCGGCGGCGGCTCCGGTGGTGGTGGTTCTCAGACTGTTGTGACTCAGGAACCTTCACTCACCGT
ATCACCTGGTGGAACAGTCACACTCACTTGTGGCTCCTCGACTGGGGCTGTTACATCTGGCTACT
ACCCAAACTGGGTCCAACAAAAACCAGGTCAGGCACCCCGTGGTCTAATAGGTGGGACTAAGTTC
CTCGCCCCCGGTACTCCTGCCAGATTCTCAGGCTCCCTGCTTGGAGGCAAGGCTGCCCTCACCCT
CTCAGGGGTACAGCCAGAGGATGAGGCAGAATATTACTGTGCTCTATGGTACAGCAACCGCTGGG
TGTTCGGTGGAGGAACCAAACTGACTGTCCTA 151. VH-VL-P of F7O artificial aa
EVQLLESGGGLVQPGGSLKLSCAASGFTFNVYAMNWVRQAPGKGLEWVARIRSKYNNYATYYADS
VKKRFTISRDDSKNTAYLQMNNLKTEDTAVYYCVRHGNFGNSYISWWAYWGQGTLVTVSSGGGGS
GGGGSGGGGSELVVTQEPSLTVSPGGTVTLTCGSSTGAVTSGYYPNWVQQKPGQAPRGLIGGTKF
LAPGTPARFSGSLLGGKAALTLSGVQPEDEAEYYCALWYSNRWVFGGGTKLTVL 152. VH-VL-P
of F7O artificial nt
GAGGTGCAGCTGCTCGAGTCTGGAGGAGGATTGGTGCAGCCTGGAGGGTCATTGAAACTCTCATG
TGCAGCCTCTGGATTCACCTTCAATGTGTACGCCATGAACTGGGTCCGCCAGGCTCCAGGAAAGG
GTTTGGAATGGGTTGCTCGCATAAGAAGTAAATATAATAATTATGCAACATATTATGCCGATTCA
GTGAAAAAGAGGTTCACCATCTCCAGAGATGATTCAAAAAACACTGCCTATCTACAAATGAACAA
CTTGAAAACTGAGGACACTGCCGTGTACTACTGTGTGAGACATGGGAACTTCGGTAATAGCTACA
TATCCTGGTGGGCTTACTGGGGCCAAGGGACTCTGGTCACCGTCTCCTCAGGTGGTGGTGGTTCT
GGCGGCGGCGGCTCCGGTGGTGGTGGTTCTGAGCTCGTTGTGACTCAGGAACCTTCACTCACCGT
ATCACCTGGTGGAACAGTCACACTCACTTGTGGCTCCTCGACTGGGGCTGTTACATCTGGCTACT
ACCCAAACTGGGTCCAACAAAAACCAGGTCAGGCACCCCGTGGTCTAATAGGTGGGACTAAGTTC
CTCGCCCCCGGTACTCCTGCCAGATTCTCAGGCTCCCTGCTTGGAGGCAAGGCTGCCCTCACCCT
CTCAGGGGTACAGCCAGAGGATGAGGCAGAATATTACTGTGCTCTATGGTACAGCAACCGCTGGG
TGTTCGGTGGAGGAACCAAACTGACTGTCCTA 153. CDR-L1 of F12Q artificial aa
GSSTGAVTSGNYPN 154. CDR-L2 of F12Q artificial aa GTKFLAP 155.
CDR-L3 of F12Q artificial aa VLWYSNRWV 156. CDR-H1 of F12Q
artificial aa SYAMN 157. CDR-H2 of F12Q artificial aa
RIRSKYNNYATYYADSVKG 158. CDR-H3 of F12Q artificial aa
HGNFGNSYVSWWAY 159. VH of F12Q artificial aa
EVQLVESGGGLVQPGGSLKLSCAASGFTFNSYAMNWVRQAPGKGLEWVARIRSKYNNYATYYADS
VKGRFTISRDDSKNTAYLQMNNLKTEDTAVYYCVRHGNFGNSYVSWWAYWGQGTLVTVSS 160.
VH of F12Q artificial nt
GAGGTGCAGCTGGTCGAGTCTGGAGGAGGATTGGTGCAGCCTGGAGGGTCATTGAAACTCTCATG
TGCAGCCTCTGGATTCACCTTCAATAGCTACGCCATGAACTGGGTCCGCCAGGCTCCAGGAAAGG
GTTTGGAATGGGTTGCTCGCATAAGAAGTAAATATAATAATTATGCAACATATTATGCCGATTCA
GTGAAAGGCAGGTTCACCATCTCCAGAGATGATTCAAAAAACACTGCCTATCTACAAATGAACAA
CTTGAAAACTGAGGACACTGCCGTGTACTACTGTGTGAGACATGGGAACTTCGGTAATAGCTACG
TTTCCTGGTGGGCTTACTGGGGCCAAGGGACTCTGGTCACCGTCTCCTCA 161. VL of F12Q
artificial aa
QTVVTQEPSLTVSPGGTVTLTCGSSTGAVTSGNYPNWVQQKPGQAPRGLIGGTKFLAPGTPARFS
GSLLGGKAALTLSGVQPEDEAEYYCVLWYSNRWVFGGGTKLTVL 162. VL of F12Q
artificial nt
CAGACTGTTGTGACTCAGGAACCTTCACTCACCGTATCACCTGGTGGAACAGTCACACTCACTTG
TGGCTCCTCGACTGGGGCTGTTACATCTGGCAACTACCCAAACTGGGTCCAACAAAAACCAGGTC
AGGCACCCCGTGGTCTAATAGGTGGGACTAAGTTCCTCGCCCCCGGTACTCCTGCCAGATTCTCA
GGCTCCCTGCTTGGAGGCAAGGCTGCCCTCACCCTCTCAGGGGTACAGCCAGAGGATGAGGCAGA
ATATTACTGTGTTCTATGGTACAGCAACCGCTGGGTGTTCGGTGGAGGAACCAAACTGACTGTCC
TA 163. VH-P of F12Q artificial aa
EVQLLESGGGLVQPGGSLKLSCAASGFTFNSYAMNWVRQAPGKGLEWVARIRSKYNNYATYYADS
VKGRFTISRDDSKNTAYLQMNNLKTEDTAVYYCVRHGNFGNSYVSWWAYWGQGTLVTVSS 164.
VH-P of F12Q artificial nt
GAGGTGCAGCTGCTCGAGTCTGGAGGAGGATTGGTGCAGCCTGGAGGGTCATTGAAACTCTCATG
TGCAGCCTCTGGATTCACCTTCAATAGCTACGCCATGAACTGGGTCCGCCAGGCTCCAGGAAAGG
GTTTGGAATGGGTTGCTCGCATAAGAAGTAAATATAATAATTATGCAACATATTATGCCGATTCA
GTGAAAGGCAGGTTCACCATCTCCAGAGATGATTCAAAAAACACTGCCTATCTACAAATGAACAA
CTTGAAAACTGAGGACACTGCCGTGTACTACTGTGTGAGACATGGGAACTTCGGTAATAGCTACG
TTTCCTGGTGGGCTTACTGGGGCCAAGGGACTCTGGTCACCGTCTCCTCA 165. VL-P of
F12Q artificial aa
ELVVTQEPSLTVSPGGTVTLTCGSSTGAVTSGNYPNWVQQKPGQAPRGLIGGTKFLAPGTPARFS
GSLLGGKAALTLSGVQPEDEAEYYCVLWYSNRWVFGGGTKLTVL 166. VL-P of F12Q
artificial nt
GAGCTCGTTGTGACTCAGGAACCTTCACTCACCGTATCACCTGGTGGAACAGTCACACTCACTTG
TGGCTCCTCGACTGGGGCTGTTACATCTGGCAACTACCCAAACTGGGTCCAACAAAAACCAGGTC
AGGCACCCCGTGGTCTAATAGGTGGGACTAAGTTCCTCGCCCCCGGTACTCCTGCCAGATTCTCA
GGCTCCCTGCTTGGAGGCAAGGCTGCCCTCACCCTCTCAGGGGTACAGCCAGAGGATGAGGCAGA
ATATTACTGTGTTCTATGGTACAGCAACCGCTGGGTGTTCGGTGGAGGAACCAAACTGACTGTCC
TA 167. VH-VL of F12Q artificial aa
EVQLVESGGGLVQPGGSLKLSCAASGFTFNSYAMNWVRQAPGKGLEWVARIRSKYNNYATYYADS
VKGRFTISRDDSKNTAYLQMNNLKTEDTAVYYCVRHGNFGNSYVSWWAYWGQGTLVTVSSGGGGS
GGGGSGGGGSQTVVTQEPSLTVSPGGTVTLTCGSSTGAVTSGNYPNWVQQKPGQAPRGLIGGTKF
LAPGTPARFSGSLLGGKAALTLSGVQPEDEAEYYCVLWYSNRWVFGGGTKLTVL 168. VH-VL
of F12Q artificial nt
GAGGTGCAGCTGGTCGAGTCTGGAGGAGGATTGGTGCAGCCTGGAGGGTCATTGAAACTCTCATG
TGCAGCCTCTGGATTCACCTTCAATAGCTACGCCATGAACTGGGTCCGCCAGGCTCCAGGAAAGG
GTTTGGAATGGGTTGCTCGCATAAGAAGTAAATATAATAATTATGCAACATATTATGCCGATTCA
GTGAAAGGCAGGTTCACCATCTCCAGAGATGATTCAAAAAACACTGCCTATCTACAAATGAACAA
CTTGAAAACTGAGGACACTGCCGTGTACTACTGTGTGAGACATGGGAACTTCGGTAATAGCTACG
TTTCCTGGTGGGCTTACTGGGGCCAAGGGACTCTGGTCACCGTCTCCTCAGGTGGTGGTGGTTCT
GGCGGCGGCGGCTCCGGTGGTGGTGGTTCTCAGACTGTTGTGACTCAGGAACCTTCACTCACCGT
ATCACCTGGTGGAACAGTCACACTCACTTGTGGCTCCTCGACTGGGGCTGTTACATCTGGCAACT
ACCCAAACTGGGTCCAACAAAAACCAGGTCAGGCACCCCGTGGTCTAATAGGTGGGACTAAGTTC
CTCGCCCCCGGTACTCCTGCCAGATTCTCAGGCTCCCTGCTTGGAGGCAAGGCTGCCCTCACCCT
CTCAGGGGTACAGCCAGAGGATGAGGCAGAATATTACTGTGTTCTATGGTACAGCAACCGCTGGG
TGTTCGGTGGAGGAACCAAACTGACTGTCCTA 169. VH-VL-P of F12Q artificial aa
EVQLLESGGGLVQPGGSLKLSCAASGFTFNSYAMNWVRQAPGKGLEWVARIRSKYNNYATYYADS
VKGRFTISRDDSKNTAYLQMNNLKTEDTAVYYCVRHGNFGNSYVSWWAYWGQGTLVTVSSGGGGS
GGGGSGGGGSELVVTQEPSLTVSPGGTVTLTCGSSTGAVTSGNYPNWVQQKPGQAPRGLIGGTKF
LAPGTPARFSGSLLGGKAALTLSGVQPEDEAEYYCVLWYSNRWVFGGGTKLTVL 170. VH-VL-P
of F12Q artificial nt
GAGGTGCAGCTGCTCGAGTCTGGAGGAGGATTGGTGCAGCCTGGAGGGTCATTGAAACTCTCATG
TGCAGCCTCTGGATTCACCTTCAATAGCTACGCCATGAACTGGGTCCGCCAGGCTCCAGGAAAGG
GTTTGGAATGGGTTGCTCGCATAAGAAGTAAATATAATAATTATGCAACATATTATGCCGATTCA
GTGAAAGGCAGGTTCACCATCTCCAGAGATGATTCAAAAAACACTGCCTATCTACAAATGAACAA
CTTGAAAACTGAGGACACTGCCGTGTACTACTGTGTGAGACATGGGAACTTCGGTAATAGCTACG
TTTCCTGGTGGGCTTACTGGGGCCAAGGGACTCTGGTCACCGTCTCCTCAGGTGGTGGTGGTTCT
GGCGGCGGCGGCTCCGGTGGTGGTGGTTCTGAGCTCGTTGTGACTCAGGAACCTTCACTCACCGT
ATCACCTGGTGGAACAGTCACACTCACTTGTGGCTCCTCGACTGGGGCTGTTACATCTGGCAACT
ACCCAAACTGGGTCCAACAAAAACCAGGTCAGGCACCCCGTGGTCTAATAGGTGGGACTAAGTTC
CTCGCCCCCGGTACTCCTGCCAGATTCTCAGGCTCCCTGCTTGGAGGCAAGGCTGCCCTCACCCT
CTCAGGGGTACAGCCAGAGGATGAGGCAGAATATTACTGTGTTCTATGGTACAGCAACCGCTGGG
TGTTCGGTGGAGGAACCAAACTGACTGTCCTA 171. CDR-L1 of I2C artificial aa
GSSTGAVTSGNYPN 172. CDR-L2 of I2C artificial aa GTKFLAP 173. CDR-L3
of I2C artificial aa VLWYSNRWV 174. CDR-H1 of I2C artificial aa
KYAMN 175. CDR-H2 of I2C artificial aa RIRSKYNNYATYYADSVKD 176.
CDR-H3 of I2C artificial aa HGNFGNSYISYWAY 177. VH of I2C
artificial aa
EVQLVESGGGLVQPGGSLKLSCAASGFTFNKYAMNWVRQAPGKGLEWVARIRSKYNNYATYYADS
VKDRFTISRDDSKNTAYLQMNNLKTEDTAVYYCVRHGNFGNSYISYWAYWGQGTLVTVSS 178.
VH of I2C artificial nt
GAGGTGCAGCTGGTCGAGTCTGGAGGAGGATTGGTGCAGCCTGGAGGGTCATTGAAACTCTCATG
TGCAGCCTCTGGATTCACCTTCAATAAGTACGCCATGAACTGGGTCCGCCAGGCTCCAGGAAAGG
GTTTGGAATGGGTTGCTCGCATAAGAAGTAAATATAATAATTATGCAACATATTATGCCGATTCA
GTGAAAGACAGGTTCACCATCTCCAGAGATGATTCAAAAAACACTGCCTATCTACAAATGAACAA
CTTGAAAACTGAGGACACTGCCGTGTACTACTGTGTGAGACATGGGAACTTCGGTAATAGCTACA
TATCCTACTGGGCTTACTGGGGCCAAGGGACTCTGGTCACCGTCTCCTCA 179. VL of I2C
artificial aa
QTVVTQEPSLTVSPGGTVTLTCGSSTGAVTSGNYPNWVQQKPGQAPRGLIGGTKFLAPGTPARFS
GSLLGGKAALTLSGVQPEDEAEYYCVLWYSNRWVFGGGTKLTVL 180. VL of I2C
artificial nt
CAGACTGTTGTGACTCAGGAACCTTCACTCACCGTATCACCTGGTGGAACAGTCACACTCACTTG
TGGCTCCTCGACTGGGGCTGTTACATCTGGCAACTACCCAAACTGGGTCCAACAAAAACCAGGTC
AGGCACCCCGTGGTCTAATAGGTGGGACTAAGTTCCTCGCCCCCGGTACTCCTGCCAGATTCTCA
GGCTCCCTGCTTGGAGGCAAGGCTGCCCTCACCCTCTCAGGGGTACAGCCAGAGGATGAGGCAGA
ATATTACTGTGTTCTATGGTACAGCAACCGCTGGGTGTTCGGTGGAGGAACCAAACTGACTGTCC
TA 181. VH-P of I2C artificial aa
EVQLLESGGGLVQPGGSLKLSCAASGFTFNKYAMNWVRQAPGKGLEWVARIRSKYNNYATYYADS
VKDRFTISRDDSKNTAYLQMNNLKTEDTAVYYCVRHGNFGNSYISYWAYWGQGTLVTVSS 182.
VH-P of I2C artificial nt
GAGGTGCAGCTGCTCGAGTCTGGAGGAGGATTGGTGCAGCCTGGAGGGTCATTGAAACTCTCATG
TGCAGCCTCTGGATTCACCTTCAATAAGTACGCCATGAACTGGGTCCGCCAGGCTCCAGGAAAGG
GTTTGGAATGGGTTGCTCGCATAAGAAGTAAATATAATAATTATGCAACATATTATGCCGATTCA
GTGAAAGACAGGTTCACCATCTCCAGAGATGATTCAAAAAACACTGCCTATCTACAAATGAACAA
CTTGAAAACTGAGGACACTGCCGTGTACTACTGTGTGAGACATGGGAACTTCGGTAATAGCTACA
TATCCTACTGGGCTTACTGGGGCCAAGGGACTCTGGTCACCGTCTCCTCA 183. VL-P of I2C
artificial aa
ELVVTQEPSLTVSPGGTVTLTCGSSTGAVTSGNYPNWVQQKPGQAPRGLIGGTKFLAPGTPARFS
GSLLGGKAALTLSGVQPEDEAEYYCVLWYSNRWVFGGGTKLTVL 184. VL-P of I2C
artificial nt
GAGCTCGTTGTGACTCAGGAACCTTCACTCACCGTATCACCTGGTGGAACAGTCACACTCACTTG
TGGCTCCTCGACTGGGGCTGTTACATCTGGCAACTACCCAAACTGGGTCCAACAAAAACCAGGTC
AGGCACCCCGTGGTCTAATAGGTGGGACTAAGTTCCTCGCCCCCGGTACTCCTGCCAGATTCTCA
GGCTCCCTGCTTGGAGGCAAGGCTGCCCTCACCCTCTCAGGGGTACAGCCAGAGGATGAGGCAGA
ATATTACTGTGTTCTATGGTACAGCAACCGCTGGGTGTTCGGTGGAGGAACCAAACTGACTGTCC
TA 185. VH-VL of I2C artificial aa
EVQLVESGGGLVQPGGSLKLSCAASGFTFNKYAMNWVRQAPGKGLEWVARIRSKYNNYATYYADS
VKDRFTISRDDSKNTAYLQMNNLKTEDTAVYYCVRHGNFGNSYISYWAYWGQGTLVTVSSGGGGS
GGGGSGGGGSQTVVTQEPSLTVSPGGTVTLTCGSSTGAVTSGNYPNWVQQKPGQAPRGLIGGTKF
LAPGTPARFSGSLLGGKAALTLSGVQPEDEAEYYCVLWYSNRWVFGGGTKLTVL 186. VH-VL
of I2C artificial nt
GAGGTGCAGCTGGTCGAGTCTGGAGGAGGATTGGTGCAGCCTGGAGGGTCATTGAAACTCTCATG
TGCAGCCTCTGGATTCACCTTCAATAAGTACGCCATGAACTGGGTCCGCCAGGCTCCAGGAAAGG
GTTTGGAATGGGTTGCTCGCATAAGAAGTAAATATAATAATTATGCAACATATTATGCCGATTCA
GTGAAAGACAGGTTCACCATCTCCAGAGATGATTCAAAAAACACTGCCTATCTACAAATGAACAA
CTTGAAAACTGAGGACACTGCCGTGTACTACTGTGTGAGACATGGGAACTTCGGTAATAGCTACA
TATCCTACTGGGCTTACTGGGGCCAAGGGACTCTGGTCACCGTCTCCTCAGGTGGTGGTGGTTCT
GGCGGCGGCGGCTCCGGTGGTGGTGGTTCTCAGACTGTTGTGACTCAGGAACCTTCACTCACCGT
ATCACCTGGTGGAACAGTCACACTCACTTGTGGCTCCTCGACTGGGGCTGTTACATCTGGCAACT
ACCCAAACTGGGTCCAACAAAAACCAGGTCAGGCACCCCGTGGTCTAATAGGTGGGACTAAGTTC
CTCGCCCCCGGTACTCCTGCCAGATTCTCAGGCTCCCTGCTTGGAGGCAAGGCTGCCCTCACCCT
CTCAGGGGTACAGCCAGAGGATGAGGCAGAATATTACTGTGTTCTATGGTACAGCAACCGCTGGG
TGTTCGGTGGAGGAACCAAACTGACTGTCCTA 187. VH-VL-P of I2C artificial aa
EVQLLESGGGLVQPGGSLKLSCAASGFTFNKYAMNWVRQAPGKGLEWVARIRSKYNNYATYYADS
VKDRFTISRDDSKNTAYLQMNNLKTEDTAVYYCVRHGNFGNSYISYWAYWGQGTLVTVSSGGGGS
GGGGSGGGGSELVVTQEPSLTVSPGGTVTLTCGSSTGAVTSGNYPNWVQQKPGQAPRGLIGGTKF
LAPGTPARFSGSLLGGKAALTLSGVQPEDEAEYYCVLWYSNRWVFGGGTKLTVL 188. VH-VL-P
of I2C artificial nt
GAGGTGCAGCTGCTCGAGTCTGGAGGAGGATTGGTGCAGCCTGGAGGGTCATTGAAACTCTCATG
TGCAGCCTCTGGATTCACCTTCAATAAGTACGCCATGAACTGGGTCCGCCAGGCTCCAGGAAAGG
GTTTGGAATGGGTTGCTCGCATAAGAAGTAAATATAATAATTATGCAACATATTATGCCGATTCA
GTGAAAGACAGGTTCACCATCTCCAGAGATGATTCAAAAAACACTGCCTATCTACAAATGAACAA
CTTGAAAACTGAGGACACTGCCGTGTACTACTGTGTGAGACATGGGAACTTCGGTAATAGCTACA
TATCCTACTGGGCTTACTGGGGCCAAGGGACTCTGGTCACCGTCTCCTCAGGTGGTGGTGGTTCT
GGCGGCGGCGGCTCCGGTGGTGGTGGTTCTGAGCTCGTTGTGACTCAGGAACCTTCACTCACCGT
ATCACCTGGTGGAACAGTCACACTCACTTGTGGCTCCTCGACTGGGGCTGTTACATCTGGCAACT
ACCCAAACTGGGTCCAACAAAAACCAGGTCAGGCACCCCGTGGTCTAATAGGTGGGACTAAGTTC
CTCGCCCCCGGTACTCCTGCCAGATTCTCAGGCTCCCTGCTTGGAGGCAAGGCTGCCCTCACCCT
CTCAGGGGTACAGCCAGAGGATGAGGCAGAATATTACTGTGTTCTATGGTACAGCAACCGCTGGG
TGTTCGGTGGAGGAACCAAACTGACTGTCCTA 189. MCSP-G4 VH-VL .times.
artificial aa
QVQLVQSGAEVKRPGASMKVSCKASGYTFTNYYIHWVRQAPGQGLEWMGWINPNSGATNYAQKFQ
H2C VH-VL
GRVTMTRDTSTSTVYMELSSLRSEDTAVYYCAKSWVSWFASWGQGTLVTVSSGGGGSGGGGSGGG
GSDIVMTQSPDSLAVSLGERATINCKSSQSVLNSSNNRNYLAWYQQKPGQPPKLLIYWASTRESG
VPDRFSGSGSGTDFTLTISGLQAEDVAVYYCQQHYSTPFTFGPGTKVDIKSGGGGSEVQLVESGG
GLVQPGGSLKLSCAASGFTFNKYAMNWVRQAPGKGLEWVARIRSKYNNYATYYADSVKDRFTISR
DDSKNTAYLQMNNLKTEDTAVYYCVRHGNFGNSYISYWAYWGQGTLVTVSSGGGGSGGGGSGGGG
SQTVVTQEPSLTVSPGGTVTLTCGSSTGAVTSGYYPNWVQQKPGQAPRGLIGGTKFLAPGTPARF
SGSLLGGKAALTLSGVQPEDEAEYYCALWYSNRWVFGGGTKLTVL 190. MCSP-G4 VH-VL
.times. artificial nt
CAGGTGCAGCTGGTCCAGTCTGGGGCTGAGGTGAAGAGGCCTGGGGCCTCAATGAAGGTCTCCTG
H2C VH-VL
CAAGGCTTCTGGGTACACCTTCACCAACTACTATATACACTGGGTGCGACAGGCCCCTGGACAAG
GTCTTGAGTGGATGGGTTGGATCAACCCTAACAGTGGTGCCACAAACTATGCACAGAAGTTCCAG
GGCAGAGTCACCATGACCAGGGACACGTCCACGAGCACAGTCTACATGGAGCTGAGCAGCCTGAG
ATCTGAGGACACGGCCGTGTATTACTGTGCGAAATCCTGGGTCTCCTGGTTTGCTTCCTGGGGTC
AAGGAACCTTGGTCACCGTCTCCTCAGGTGGTGGTGGTTCTGGCGGCGGCGGCTCCGGTGGTGGT
GGTTCTGATATCGTGATGACCCAGTCTCCAGACTCCCTGGCTGTGTCTCTGGGCGAGAGGGCCAC
CATCAACTGCAAGTCCAGCCAGAGTGTTTTAAACAGCTCCAACAATAGGAACTACTTAGCTTGGT
ACCAGCAGAAACCAGGACAGCCTCCTAAGCTGCTCATTTACTGGGCATCTACCCGGGAATCCGGG
GTCCCTGACCGATTCAGTGGCAGCGGGTCTGGGACAGATTTCACTCTCACCATCAGTGGCCTGCA
GGCTGAAGATGTGGCAGTTTATTACTGTCAGCAACATTATAGTACTCCATTCACTTTTGGCCCTG
GGACCAAAGTGGATATCAAATCCGGAGGTGGTGGATCCGAGGTGCAGCTGGTCGAGTCTGGAGGA
GGATTGGTGCAGCCTGGAGGGTCATTGAAACTCTCATGTGCAGCCTCTGGATTCACCTTCAATAA
GTACGCCATGAACTGGGTCCGCCAGGCTCCAGGAAAGGGTTTGGAATGGGTTGCTCGCATAAGAA
GTAAATATAATAATTATGCAACATATTATGCCGATTCAGTGAAAGACAGGTTCACCATCTCCAGA
GATGATTCAAAAAACACTGCCTATCTACAAATGAACAACTTGAAAACTGAGGACACTGCCGTGTA
CTACTGTGTGAGACATGGGAACTTCGGTAATAGCTACATATCCTACTGGGCTTACTGGGGCCAAG
GGACTCTGGTCACCGTCTCCTCAGGTGGTGGTGGTTCTGGCGGCGGCGGCTCCGGTGGTGGTGGT
TCTCAGACTGTTGTGACTCAGGAACCTTCACTCACCGTATCACCTGGTGGAACAGTCACACTCAC
TTGTGGCTCCTCGACTGGGGCTGTTACATCTGGCTACTACCCAAACTGGGTCCAACAAAAACCAG
GTCAGGCACCCCGTGGTCTAATAGGTGGGACTAAGTTCCTCGCCCCCGGTACTCCTGCCAGATTC
TCAGGCTCCCTGCTTGGAGGCAAGGCTGCCCTCACCCTCTCAGGGGTACAGCCAGAGGATGAGGC
AGAATATTACTGTGCTCTATGGTACAGCAACCGCTGGGTGTTCGGTGGAGGAACCAAACTGACTG
TCCTA 191. MCSP-G4 VH-VL .times. artificial aa
QVQLVQSGAEVKRPGASMKVSCKASGYTFTNYYIHWVRQAPGQGLEWMGWINPNSGATNYAQKFQ
F12Q VH-VL
GRVTMTRDTSTSTVYMELSSLRSEDTAVYYCAKSWVSWFASWGQGTLVTVSSGGGGSGGGGSGGG
GSDIVMTQSPDSLAVSLGERATINCKSSQSVLNSSNNRNYLAWYQQKPGQPPKLLIYWASTRESG
VPDRFSGSGSGTDFTLTISGLQAEDVAVYYCQQHYSTPFTFGPGTKVDIKSGGGGSEVQLVESGG
GLVQPGGSLKLSCAASGFTFNSYAMNWVRQAPGKGLEWVARIRSKYNNYATYYADSVKGRFTISR
DDSKNTAYLQMNNLKTEDTAVYYCVRHGNFGNSYVSWWAYWGQGTLVTVSSGGGGSGGGGSGGGG
SQTVVTQEPSLTVSPGGTVTLTCGSSTGAVTSGNYPNWVQQKPGQAPRGLIGGTKFLAPGTPARF
SGSLLGGKAALTLSGVQPEDEAEYYCVLWYSNRWVFGGGTKLTVL 192. MCSP-G4 VH-VL
.times. artificial nt
CAGGTGCAGCTGGTCCAGTCTGGGGCTGAGGTGAAGAGGCCTGGGGCCTCAATGAAGGTCTCCTG
F12Q VH-VL
CAAGGCTTCTGGGTACACCTTCACCAACTACTATATACACTGGGTGCGACAGGCCCCTGGACAAG
GTCTTGAGTGGATGGGTTGGATCAACCCTAACAGTGGTGCCACAAACTATGCACAGAAGTTCCAG
GGCAGAGTCACCATGACCAGGGACACGTCCACGAGCACAGTCTACATGGAGCTGAGCAGCCTGAG
ATCTGAGGACACGGCCGTGTATTACTGTGCGAAATCCTGGGTCTCCTGGTTTGCTTCCTGGGGTC
AAGGAACCTTGGTCACCGTCTCCTCAGGTGGTGGTGGTTCTGGCGGCGGCGGCTCCGGTGGTGGT
GGTTCTGATATCGTGATGACCCAGTCTCCAGACTCCCTGGCTGTGTCTCTGGGCGAGAGGGCCAC
CATCAACTGCAAGTCCAGCCAGAGTGTTTTAAACAGCTCCAACAATAGGAACTACTTAGCTTGGT
ACCAGCAGAAACCAGGACAGCCTCCTAAGCTGCTCATTTACTGGGCATCTACCCGGGAATCCGGG
GTCCCTGACCGATTCAGTGGCAGCGGGTCTGGGACAGATTTCACTCTCACCATCAGTGGCCTGCA
GGCTGAAGATGTGGCAGTTTATTACTGTCAGCAACATTATAGTACTCCATTCACTTTTGGCCCTG
GGACCAAAGTGGATATCAAATCCGGAGGTGGTGGATCCGAGGTGCAGCTGGTCGAGTCTGGAGGA
GGATTGGTGCAGCCTGGAGGGTCATTGAAACTCTCATGTGCAGCCTCTGGATTCACCTTCAATAG
CTACGCCATGAACTGGGTCCGCCAGGCTCCAGGAAAGGGTTTGGAATGGGTTGCTCGCATAAGAA
GTAAATATAATAATTATGCAACATATTATGCCGATTCAGTGAAAGGCAGGTTCACCATCTCCAGA
GATGATTCAAAAAACACTGCCTATCTACAAATGAACAACTTGAAAACTGAGGACACTGCCGTGTA
CTACTGTGTGAGACATGGGAACTTCGGTAATAGCTACGTTTCCTGGTGGGCTTACTGGGGCCAAG
GGACTCTGGTCACCGTCTCCTCAGGTGGTGGTGGTTCTGGCGGCGGCGGCTCCGGTGGTGGTGGT
TCTCAGACTGTTGTGACTCAGGAACCTTCACTCACCGTATCACCTGGTGGAACAGTCACACTCAC
TTGTGGCTCCTCGACTGGGGCTGTTACATCTGGCAACTACCCAAACTGGGTCCAACAAAAACCAG
GTCAGGCACCCCGTGGTCTAATAGGTGGGACTAAGTTCCTCGCCCCCGGTACTCCTGCCAGATTC
TCAGGCTCCCTGCTTGGAGGCAAGGCTGCCCTCACCCTCTCAGGGGTACAGCCAGAGGATGAGGC
AGAATATTACTGTGTTCTATGGTACAGCAACCGCTGGGTGTTCGGTGGAGGAACCAAACTGACTG
TCCTA 193. MCSP-G4 VH-VL .times. artificial aa
QVQLVQSGAEVKRPGASMKVSCKASGYTFTNYYIHWVRQAPGQGLEWMGWINPNSGATNYAQKFQ
I2C VH-VL
GRVTMTRDTSTSTVYMELSSLRSEDTAVYYCAKSWVSWFASWGQGTLVTVSSGGGGSGGGGSGGG
GSDIVMTQSPDSLAVSLGERATINCKSSQSVLNSSNNRNYLAWYQQKPGQPPKLLIYWASTRESG
VPDRFSGSGSGTDFTLTISGLQAEDVAVYYCQQHYSTPFTFGPGTKVDIKSGGGGSEVQLVESGG
GLVQPGGSLKLSCAASGFTFNKYAMNWVRQAPGKGLEWVARIRSKYNNYATYYADSVKDRFTISR
DDSKNTAYLQMNNLKTEDTAVYYCVRHGNFGNSYISYWAYWGQGTLVTVSSGGGGSGGGGSGGGG
SQTVVTQEPSLTVSPGGTVTLTCGSSTGAVTSGNYPNWVQQKPGQAPRGLIGGTKFLAPGTPARF
SGSLLGGKAALTLSGVQPEDEAEYYCVLWYSNRWVFGGGTKLTVL 194. MCSP-G4 VH-VL
.times. artificial nt
CAGGTGCAGCTGGTCCAGTCTGGGGCTGAGGTGAAGAGGCCTGGGGCCTCAATGAAGGTCTCCTG
I2C VH-VL
CAAGGCTTCTGGGTACACCTTCACCAACTACTATATACACTGGGTGCGACAGGCCCCTGGACAAG
GTCTTGAGTGGATGGGTTGGATCAACCCTAACAGTGGTGCCACAAACTATGCACAGAAGTTCCAG
GGCAGAGTCACCATGACCAGGGACACGTCCACGAGCACAGTCTACATGGAGCTGAGCAGCCTGAG
ATCTGAGGACACGGCCGTGTATTACTGTGCGAAATCCTGGGTCTCCTGGTTTGCTTCCTGGGGTC
AAGGAACCTTGGTCACCGTCTCCTCAGGTGGTGGTGGTTCTGGCGGCGGCGGCTCCGGTGGTGGT
GGTTCTGATATCGTGATGACCCAGTCTCCAGACTCCCTGGCTGTGTCTCTGGGCGAGAGGGCCAC
CATCAACTGCAAGTCCAGCCAGAGTGTTTTAAACAGCTCCAACAATAGGAACTACTTAGCTTGGT
ACCAGCAGAAACCAGGACAGCCTCCTAAGCTGCTCATTTACTGGGCATCTACCCGGGAATCCGGG
GTCCCTGACCGATTCAGTGGCAGCGGGTCTGGGACAGATTTCACTCTCACCATCAGTGGCCTGCA
GGCTGAAGATGTGGCAGTTTATTACTGTCAGCAACATTATAGTACTCCATTCACTTTTGGCCCTG
GGACCAAAGTGGATATCAAATCCGGAGGTGGTGGATCCGAGGTGCAGCTGGTCGAGTCTGGAGGA
GGATTGGTGCAGCCTGGAGGGTCATTGAAACTCTCATGTGCAGCCTCTGGATTCACCTTCAATAA
GTACGCCATGAACTGGGTCCGCCAGGCTCCAGGAAAGGGTTTGGAATGGGTTGCTCGCATAAGAA
GTAAATATAATAATTATGCAACATATTATGCCGATTCAGTGAAAGACAGGTTCACCATCTCCAGA
GATGATTCAAAAAACACTGCCTATCTACAAATGAACAACTTGAAAACTGAGGACACTGCCGTGTA
CTACTGTGTGAGACATGGGAACTTCGGTAATAGCTACATATCCTACTGGGCTTACTGGGGCCAAG
GGACTCTGGTCACCGTCTCCTCAGGTGGTGGTGGTTCTGGCGGCGGCGGCTCCGGTGGTGGTGGT
TCTCAGACTGTTGTGACTCAGGAACCTTCACTCACCGTATCACCTGGTGGAACAGTCACACTCAC
TTGTGGCTCCTCGACTGGGGCTGTTACATCTGGCAACTACCCAAACTGGGTCCAACAAAAACCAG
GTCAGGCACCCCGTGGTCTAATAGGTGGGACTAAGTTCCTCGCCCCCGGTACTCCTGCCAGATTC
TCAGGCTCCCTGCTTGGAGGCAAGGCTGCCCTCACCCTCTCAGGGGTACAGCCAGAGGATGAGGC
AGAATATTACTGTGTTCTATGGTACAGCAACCGCTGGGTGTTCGGTGGAGGAACCAAACTGACTG
TCCTA 195. MCSP-G4 VH-VL-P .times. artificial aa
QVQLVQSGAEVKRPGASMKVSCKASGYTFTNYYIHWVRQAPGQGLEWMGWINPNSGATNYAQKFQ
F6A VH-VL-P
GRVTMTRDTSTSTVYMELSSLRSEDTAVYYCAKSWVSWFASWGQGTLVTVSSGGGGSGGGGSGGG
GSELVMTQSPDSLAVSLGERATINCKSSQSVLNSSNNRNYLAWYQQKPGQPPKLLIYWASTRESG
VPDRFSGSGSGTDFTLTISGLQAEDVAVYYCQQHYSTPFTFGPGTKVDIKSGGGGSEVQLLESGG
GLVQPGGSLKLSCAASGFTFNIYAMNWVRQAPGKGLEWVARIRSKYNNYATYYADSVKSRFTISR
DDSKNTAYLQMNNLKTEDTAVYYCVRHGNFGNSYVSFFAYWGQGTLVTVSSGGGGSGGGGSGGGG
SELVVTQEPSLTVSPGGTVTLTCGSSTGAVTSGYYPNWVQQKPGQAPRGLIGGTKFLAPGTPARF
SGSLLGGKAALTLSGVQPEDEAEYYCALWYSNRWVFGGGTKLTVL 196. MCSP-G4 VH-VL-P
.times. artificial nt
CAGGTGCAGCTGGTCCAGTCTGGGGCTGAGGTGAAGAGGCCTGGGGCCTCAATGAAGGTCTCCTG
F6A VH-VL-P
CAAGGCTTCTGGGTACACCTTCACCAACTACTATATACACTGGGTGCGACAGGCCCCTGGACAAG
GTCTTGAGTGGATGGGTTGGATCAACCCTAACAGTGGTGCCACAAACTATGCACAGAAGTTCCAG
GGCAGAGTCACCATGACCAGGGACACGTCCACGAGCACAGTCTACATGGAGCTGAGCAGCCTGAG
ATCTGAGGACACGGCCGTGTATTACTGTGCGAAATCCTGGGTCTCCTGGTTTGCTTCCTGGGGTC
AAGGAACCTTGGTCACCGTCTCCTCAGGTGGTGGTGGTTCTGGCGGCGGCGGCTCCGGTGGTGGT
GGTTCTGAGCTCGTGATGACCCAGTCTCCAGACTCCCTGGCTGTGTCTCTGGGCGAGAGGGCCAC
CATCAACTGCAAGTCCAGCCAGAGTGTTTTAAACAGCTCCAACAATAGGAACTACTTAGCTTGGT
ACCAGCAGAAACCAGGACAGCCTCCTAAGCTGCTCATTTACTGGGCATCTACCCGGGAATCCGGG
GTCCCTGACCGATTCAGTGGCAGCGGGTCTGGGACAGATTTCACTCTCACCATCAGTGGCCTGCA
GGCTGAAGATGTGGCAGTTTATTACTGTCAGCAACATTATAGTACTCCATTCACTTTTGGCCCTG
GGACCAAAGTGGATATCAAATCCGGAGGTGGTGGATCCGAGGTGCAGCTGCTCGAGTCTGGAGGA
GGATTGGTGCAGCCTGGAGGGTCATTGAAACTCTCATGTGCAGCCTCTGGATTCACCTTCAATAT
CTACGCCATGAACTGGGTCCGCCAGGCTCCAGGAAAGGGTTTGGAATGGGTTGCTCGCATAAGAA
GTAAATATAATAATTATGCAACATATTATGCCGATTCAGTGAAAAGCAGGTTCACCATCTCCAGA
GATGATTCAAAAAACACTGCCTATCTACAAATGAACAACTTGAAAACTGAGGACACTGCCGTGTA
CTACTGTGTGAGACATGGGAACTTCGGTAATAGCTACGTATCCTTCTTCGCTTACTGGGGCCAAG
GGACTCTGGTCACCGTCTCCTCAGGTGGTGGTGGTTCTGGCGGCGGCGGCTCCGGTGGTGGTGGT
TCTGAGCTCGTTGTGACTCAGGAACCTTCACTCACCGTATCACCTGGTGGAACAGTCACACTCAC
TTGTGGCTCCTCGACTGGGGCTGTTACATCTGGCTACTACCCAAACTGGGTCCAACAAAAACCAG
GTCAGGCACCCCGTGGTCTAATAGGTGGGACTAAGTTCCTCGCCCCCGGTACTCCTGCCAGATTC
TCAGGCTCCCTGCTTGGAGGCAAGGCTGCCCTCACCCTCTCAGGGGTACAGCCAGAGGATGAGGC
AGAATATTACTGTGCTCTATGGTACAGCAACCGCTGGGTGTTCGGTGGAGGAACCAAACTGACTG
TCCTA 197. MCSP-G4 VH-VL-P .times. artificial aa
QVQLVQSGAEVKRPGASMKVSCKASGYTFTNYYIHWVRQAPGQGLEWMGWINPNSGATNYAQKFQ
H2C VH-VL-P
GRVTMTRDTSTSTVYMELSSLRSEDTAVYYCAKSWVSWFASWGQGTLVTVSSGGGGSGGGGSGGG
GSELVMTQSPDSLAVSLGERATINCKSSQSVLNSSNNRNYLAWYQQKPGQPPKLLIYWASTRESG
VPDRFSGSGSGTDFTLTISGLQAEDVAVYYCQQHYSTPFTFGPGTKVDIKSGGGGSEVQLLESGG
GLVQPGGSLKLSCAASGFTFNKYAMNWVRQAPGKGLEWVARIRSKYNNYATYYADSVKDRFTISR
DDSKNTAYLQMNNLKTEDTAVYYCVRHGNFGNSYISYWAYWGQGTLVTVSSGGGGSGGGGSGGGG
SELVVTQEPSLTVSPGGTVTLTCGSSTGAVTSGYYPNWVQQKPGQAPRGLIGGTKFLAPGTPARF
SGSLLGGKAALTLSGVQPEDEAEYYCALWYSNRWVFGGGTKLTVL 198. MCSP-G4 VH-VL-P
.times. artificial nt
CAGGTGCAGCTGGTCCAGTCTGGGGCTGAGGTGAAGAGGCCTGGGGCCTCAATGAAGGTCTCCTG
H2C VH-VL-P
CAAGGCTTCTGGGTACACCTTCACCAACTACTATATACACTGGGTGCGACAGGCCCCTGGACAAG
GTCTTGAGTGGATGGGTTGGATCAACCCTAACAGTGGTGCCACAAACTATGCACAGAAGTTCCAG
GGCAGAGTCACCATGACCAGGGACACGTCCACGAGCACAGTCTACATGGAGCTGAGCAGCCTGAG
ATCTGAGGACACGGCCGTGTATTACTGTGCGAAATCCTGGGTCTCCTGGTTTGCTTCCTGGGGTC
AAGGAACCTTGGTCACCGTCTCCTCAGGTGGTGGTGGTTCTGGCGGCGGCGGCTCCGGTGGTGGT
GGTTCTGAGCTCGTGATGACCCAGTCTCCAGACTCCCTGGCTGTGTCTCTGGGCGAGAGGGCCAC
CATCAACTGCAAGTCCAGCCAGAGTGTTTTAAACAGCTCCAACAATAGGAACTACTTAGCTTGGT
ACCAGCAGAAACCAGGACAGCCTCCTAAGCTGCTCATTTACTGGGCATCTACCCGGGAATCCGGG
GTCCCTGACCGATTCAGTGGCAGCGGGTCTGGGACAGATTTCACTCTCACCATCAGTGGCCTGCA
GGCTGAAGATGTGGCAGTTTATTACTGTCAGCAACATTATAGTACTCCATTCACTTTTGGCCCTG
GGACCAAAGTGGATATCAAATCCGGAGGTGGTGGATCCGAGGTGCAGCTGCTCGAGTCTGGAGGA
GGATTGGTGCAGCCTGGAGGGTCATTGAAACTCTCATGTGCAGCCTCTGGATTCACCTTCAATAA
GTACGCCATGAACTGGGTCCGCCAGGCTCCAGGAAAGGGTTTGGAATGGGTTGCTCGCATAAGAA
GTAAATATAATAATTATGCAACATATTATGCCGATTCAGTGAAAGACAGGTTCACCATCTCCAGA
GATGATTCAAAAAACACTGCCTATCTACAAATGAACAACTTGAAAACTGAGGACACTGCCGTGTA
CTACTGTGTGAGACATGGGAACTTCGGTAATAGCTACATATCCTACTGGGCTTACTGGGGCCAAG
GGACTCTGGTCACCGTCTCCTCAGGTGGTGGTGGTTCTGGCGGCGGCGGCTCCGGTGGTGGTGGT
TCTGAGCTCGTTGTGACTCAGGAACCTTCACTCACCGTATCACCTGGTGGAACAGTCACACTCAC
TTGTGGCTCCTCGACTGGGGCTGTTACATCTGGCTACTACCCAAACTGGGTCCAACAAAAACCAG
GTCAGGCACCCCGTGGTCTAATAGGTGGGACTAAGTTCCTCGCCCCCGGTACTCCTGCCAGATTC
TCAGGCTCCCTGCTTGGAGGCAAGGCTGCCCTCACCCTCTCAGGGGTACAGCCAGAGGATGAGGC
AGAATATTACTGTGCTCTATGGTACAGCAACCGCTGGGTGTTCGGTGGAGGAACCAAACTGACTG
TCCTA 199. MCSP-G4 VH-VL-P .times. artificial aa
QVQLVQSGAEVKRPGASMKVSCKASGYTFTNYYIHWVRQAPGQGLEWMGWINPNSGATNYAQKFQ
H1E VH-VL-P
GRVTMTRDTSTSTVYMELSSLRSEDTAVYYCAKSWVSWFASWGQGTLVTVSSGGGGSGGGGSGGG
GSELVMTQSPDSLAVSLGERATINCKSSQSVLNSSNNRNYLAWYQQKPGQPPKLLIYWASTRESG
VPDRFSGSGSGTDFTLTISGLQAEDVAVYYCQQHYSTPFTFGPGTKVDIKSGGGGSEVQLLESGG
GLEQPGGSLKLSCAASGFTFNSYAMNWVRQAPGKGLEWVARIRSKYNNYATYYADSVKGRFTISR
DDSKNTAYLQMNNLKTEDTAVYYCVRHGNFGNSYLSFWAYWGQGTLVTVSSGGGGSGGGGSGGGG
SELVVTQEPSLTVSPGGTVTLTCGSSTGAVTSGYYPNWVQQKPGQAPRGLIGGTKFLAPGTPARF
SGSLLGGKAALTLSGVQPEDEAEYYCALWYSNRWVFGGGTKLTVL 200. MCSP-G4 VH-VL-P
.times. artificial nt
CAGGTGCAGCTGGTCCAGTCTGGGGCTGAGGTGAAGAGGCCTGGGGCCTCAATGAAGGTCTCCTG
H1E VH-VL-P
CAAGGCTTCTGGGTACACCTTCACCAACTACTATATACACTGGGTGCGACAGGCCCCTGGACAAG
GTCTTGAGTGGATGGGTTGGATCAACCCTAACAGTGGTGCCACAAACTATGCACAGAAGTTCCAG
GGCAGAGTCACCATGACCAGGGACACGTCCACGAGCACAGTCTACATGGAGCTGAGCAGCCTGAG
ATCTGAGGACACGGCCGTGTATTACTGTGCGAAATCCTGGGTCTCCTGGTTTGCTTCCTGGGGTC
AAGGAACCTTGGTCACCGTCTCCTCAGGTGGTGGTGGTTCTGGCGGCGGCGGCTCCGGTGGTGGT
GGTTCTGAGCTCGTGATGACCCAGTCTCCAGACTCCCTGGCTGTGTCTCTGGGCGAGAGGGCCAC
CATCAACTGCAAGTCCAGCCAGAGTGTTTTAAACAGCTCCAACAATAGGAACTACTTAGCTTGGT
ACCAGCAGAAACCAGGACAGCCTCCTAAGCTGCTCATTTACTGGGCATCTACCCGGGAATCCGGG
GTCCCTGACCGATTCAGTGGCAGCGGGTCTGGGACAGATTTCACTCTCACCATCAGTGGCCTGCA
GGCTGAAGATGTGGCAGTTTATTACTGTCAGCAACATTATAGTACTCCATTCACTTTTGGCCCTG
GGACCAAAGTGGATATCAAATCCGGAGGTGGTGGATCCGAGGTGCAGCTGCTCGAGTCTGGAGGA
GGATTGGAGCAGCCTGGAGGGTCATTGAAACTCTCATGTGCAGCCTCTGGATTCACCTTCAATTC
GTACGCCATGAACTGGGTCCGCCAGGCTCCAGGAAAGGGTTTGGAATGGGTTGCTCGCATAAGAA
GTAAATATAATAATTATGCAACATATTATGCCGATTCAGTGAAAGGGAGGTTCACCATCTCCAGA
GATGATTCAAAAAACACTGCCTATCTACAAATGAACAACTTGAAAACTGAGGACACTGCCGTGTA
CTACTGTGTGAGACATGGGAACTTCGGTAATAGCTACCTATCCTTCTGGGCTTACTGGGGCCAAG
GGACTCTGGTCACCGTCTCCTCAGGTGGTGGTGGTTCTGGCGGCGGCGGCTCCGGTGGTGGTGGT
TCTGAGCTCGTTGTGACTCAGGAACCTTCACTCACCGTATCACCTGGTGGAACAGTCACACTCAC
TTGTGGCTCCTCGACTGGGGCTGTTACATCTGGCTACTACCCAAACTGGGTCCAACAAAAACCAG
GTCAGGCACCCCGTGGTCTAATAGGTGGGACTAAGTTCCTCGCCCCCGGTACTCCTGCCAGATTC
TCAGGCTCCCTGCTTGGAGGCAAGGCTGCCCTCACCCTCTCAGGGGTACAGCCAGAGGATGAGGC
AGAATATTACTGTGCTCTATGGTACAGCAACCGCTGGGTGTTCGGTGGAGGAACCAAACTGACTG
TCCTA 201. MCSP-G4 VH-VL-P .times. artificial aa
QVQLVQSGAEVKRPGASMKVSCKASGYTFTNYYIHWVRQAPGQGLEWMGWINPNSGATNYAQKFQ
G4H VH-VL-P
GRVTMTRDTSTSTVYMELSSLRSEDTAVYYCAKSWVSWFASWGQGTLVTVSSGGGGSGGGGSGGG
GSELVMTQSPDSLAVSLGERATINCKSSQSVLNSSNNRNYLAWYQQKPGQPPKLLIYWASTRESG
VPDRFSGSGSGTDFTLTISGLQAEDVAVYYCQQHYSTPFTFGPGTKVDIKSGGGGSEVQLLESGG
GLVQPGGSLKLSCAASGFTFNRYAMNWVRQAPGKGLEWVARIRSKYNNYATYYADSVKGRFTISR
DDSKNTAYLQMNNLKTEDTAVYYCVRHGNFGNSYLSYFAYWGQGTLVTVSSGGGGSGGGGSGGGG
SELVVTQEPSLTVSPGGTVTLTCGSSTGAVTSGYYPNWVQQKPGQAPRGLIGGTKFLAPGTPARF
SGSLLGGKAALTLSGVQPEDEAEYYCALWYSNRWVFGGGTKLTVL 202. MCSP-G4 VH-VL-P
.times. artificial nt
CAGGTGCAGCTGGTCCAGTCTGGGGCTGAGGTGAAGAGGCCTGGGGCCTCAATGAAGGTCTCCTG
G4H VH-VL-P
CAAGGCTTCTGGGTACACCTTCACCAACTACTATATACACTGGGTGCGACAGGCCCCTGGACAAG
GTCTTGAGTGGATGGGTTGGATCAACCCTAACAGTGGTGCCACAAACTATGCACAGAAGTTCCAG
GGCAGAGTCACCATGACCAGGGACACGTCCACGAGCACAGTCTACATGGAGCTGAGCAGCCTGAG
ATCTGAGGACACGGCCGTGTATTACTGTGCGAAATCCTGGGTCTCCTGGTTTGCTTCCTGGGGTC
AAGGAACCTTGGTCACCGTCTCCTCAGGTGGTGGTGGTTCTGGCGGCGGCGGCTCCGGTGGTGGT
GGTTCTGAGCTCGTGATGACCCAGTCTCCAGACTCCCTGGCTGTGTCTCTGGGCGAGAGGGCCAC
CATCAACTGCAAGTCCAGCCAGAGTGTTTTAAACAGCTCCAACAATAGGAACTACTTAGCTTGGT
ACCAGCAGAAACCAGGACAGCCTCCTAAGCTGCTCATTTACTGGGCATCTACCCGGGAATCCGGG
GTCCCTGACCGATTCAGTGGCAGCGGGTCTGGGACAGATTTCACTCTCACCATCAGTGGCCTGCA
GGCTGAAGATGTGGCAGTTTATTACTGTCAGCAACATTATAGTACTCCATTCACTTTTGGCCCTG
GGACCAAAGTGGATATCAAATCCGGAGGTGGTGGATCCGAGGTGCAGCTGCTCGAGTCTGGAGGA
GGATTGGTGCAGCCTGGAGGGTCATTGAAACTCTCATGTGCAGCCTCTGGATTCACCTTCAATCG
CTACGCCATGAACTGGGTCCGCCAGGCTCCAGGAAAGGGTTTGGAATGGGTTGCTCGCATAAGAA
GTAAATATAATAATTATGCAACATATTATGCCGATTCAGTGAAAGGGAGGTTCACCATCTCCAGA
GATGATTCAAAAAACACTGCCTATCTACAAATGAACAACTTGAAAACTGAGGACACTGCCGTGTA
CTACTGTGTGAGACATGGGAACTTCGGTAATAGCTACTTATCCTACTTCGCTTACTGGGGCCAAG
GGACTCTGGTCACCGTCTCCTCAGGTGGTGGTGGTTCTGGCGGCGGCGGCTCCGGTGGTGGTGGT
TCTGAGCTCGTTGTGACTCAGGAACCTTCACTCACCGTATCACCTGGTGGAACAGTCACACTCAC
TTGTGGCTCCTCGACTGGGGCTGTTACATCTGGCTACTACCCAAACTGGGTCCAACAAAAACCAG
GTCAGGCACCCCGTGGTCTAATAGGTGGGACTAAGTTCCTCGCCCCCGGTACTCCTGCCAGATTC
TCAGGCTCCCTGCTTGGAGGCAAGGCTGCCCTCACCCTCTCAGGGGTACAGCCAGAGGATGAGGC
AGAATATTACTGTGCTCTATGGTACAGCAACCGCTGGGTGTTCGGTGGAGGAACCAAACTGACTG
TCCTA 203. MCSP-G4 VH-VL-P .times. artificial aa
QVQLVQSGAEVKRPGASMKVSCKASGYTFTNYYIHWVRQAPGQGLEWMGWINPNSGATNYAQKFQ
A2J VH-VL-P
GRVTMTRDTSTSTVYMELSSLRSEDTAVYYCAKSWVSWFASWGQGTLVTVSSGGGGSGGGGSGGG
GSELVMTQSPDSLAVSLGERATINCKSSQSVLNSSNNRNYLAWYQQKPGQPPKLLIYWASTRESG
VPDRFSGSGSGTDFTLTISGLQAEDVAVYYCQQHYSTPFTFGPGTKVDIKSGGGGSEVQLLESGG
GLVQPGGSLKLSCAASGFTFNVYAMNWVRQAPGKGLEWVARIRSKYNNYATYYADSVKKRFTISR
DDSKNTAYLQMNNLKTEDTAVYYCVRHGNFGNSYLSWWAYWGQGTLVTVSSGGGGSGGGGSGGGG
SELVVTQEPSLTVSPGGTVTLTCRSSTGAVTSGYYPNWVQQKPGQAPRGLIGATDMRPSGTPARF
SGSLLGGKAALTLSGVQPEDEAEYYCALWYSNRWVFGGGTKLTVL 204. MCSP-G4 VH-VL-P
.times. artificial nt
CAGGTGCAGCTGGTCCAGTCTGGGGCTGAGGTGAAGAGGCCTGGGGCCTCAATGAAGGTCTCCTG
A2J VH-VL-P
CAAGGCTTCTGGGTACACCTTCACCAACTACTATATACACTGGGTGCGACAGGCCCCTGGACAAG
GTCTTGAGTGGATGGGTTGGATCAACCCTAACAGTGGTGCCACAAACTATGCACAGAAGTTCCAG
GGCAGAGTCACCATGACCAGGGACACGTCCACGAGCACAGTCTACATGGAGCTGAGCAGCCTGAG
ATCTGAGGACACGGCCGTGTATTACTGTGCGAAATCCTGGGTCTCCTGGTTTGCTTCCTGGGGTC
AAGGAACCTTGGTCACCGTCTCCTCAGGTGGTGGTGGTTCTGGCGGCGGCGGCTCCGGTGGTGGT
GGTTCTGAGCTCGTGATGACCCAGTCTCCAGACTCCCTGGCTGTGTCTCTGGGCGAGAGGGCCAC
CATCAACTGCAAGTCCAGCCAGAGTGTTTTAAACAGCTCCAACAATAGGAACTACTTAGCTTGGT
ACCAGCAGAAACCAGGACAGCCTCCTAAGCTGCTCATTTACTGGGCATCTACCCGGGAATCCGGG
GTCCCTGACCGATTCAGTGGCAGCGGGTCTGGGACAGATTTCACTCTCACCATCAGTGGCCTGCA
GGCTGAAGATGTGGCAGTTTATTACTGTCAGCAACATTATAGTACTCCATTCACTTTTGGCCCTG
GGACCAAAGTGGATATCAAATCCGGAGGTGGTGGATCCGAGGTGCAGCTGCTCGAGTCTGGAGGA
GGATTGGTGCAGCCTGGAGGGTCATTGAAACTCTCATGTGCAGCCTCTGGATTCACCTTCAATGT
CTACGCCATGAACTGGGTCCGCCAGGCTCCAGGAAAGGGTTTGGAATGGGTTGCTCGCATAAGAA
GTAAATATAATAATTATGCAACATATTATGCCGATTCAGTGAAAAAGAGGTTCACCATCTCCAGA
GATGATTCAAAAAACACTGCCTATCTACAAATGAACAACTTGAAAACTGAGGACACTGCCGTGTA
CTACTGTGTGAGACATGGGAACTTCGGTAATAGCTACTTATCCTGGTGGGCTTACTGGGGCCAAG
GGACTCTGGTCACCGTCTCCTCAGGTGGTGGTGGTTCTGGCGGCGGCGGCTCCGGTGGTGGTGGT
TCTGAGCTCGTTGTGACTCAGGAACCTTCACTCACCGTATCACCTGGTGGAACAGTCACACTCAC
TTGTCGCTCCTCGACTGGGGCTGTTACATCTGGCTACTACCCAAACTGGGTCCAACAAAAACCAG
GTCAGGCACCCCGTGGTCTAATAGGTGCCACTGACATGAGGCCCTCTGGTACTCCTGCCAGATTC
TCAGGCTCCCTGCTTGGAGGCAAGGCTGCCCTCACCCTCTCAGGGGTACAGCCAGAGGATGAGGC
AGAATATTACTGTGCTCTATGGTACAGCAACCGCTGGGTGTTCGGTGGAGGAACCAAACTGACTG
TCCTA 205. MCSP-G4 VH-VL-P .times. artificial aa
QVQLVQSGAEVKRPGASMKVSCKASGYTFTNYYIHWVRQAPGQGLEWMGWINPNSGATNYAQKFQ
E1L VH-VL-P
GRVTMTRDTSTSTVYMELSSLRSEDTAVYYCAKSWVSWFASWGQGTLVTVSSGGGGSGGGGSGGG
GSELVMTQSPDSLAVSLGERATINCKSSQSVLNSSNNRNYLAWYQQKPGQPPKLLIYWASTRESG
VPDRFSGSGSGTDFTLTISGLQAEDVAVYYCQQHYSTPFTFGPGTKVDIKSGGGGSEVQLLESGG
GLVQPGGSLKLSCAASGFTFNKYAMNWVRQAPGKGLEWVARIRSKYNNYATYYADSVKSRFTISR
DDSKNTAYLQMNNLKTEDTAVYYCVRHGNFGNSYTSYYAYWGQGTLVTVSSGGGGSGGGGSGGGG
SELVVTQEPSLTVSPGGTVTLTCGSSTGAVTSGYYPNWVQQKPGQAPRGLIGGTKFLAPGTPARF
SGSLLGGKAALTLSGVQPEDEAEYYCALWYSNRWVFGGGTKLTVL 206. MCSP-G4 VH-VL-P
.times. artificial nt
CAGGTGCAGCTGGTCCAGTCTGGGGCTGAGGTGAAGAGGCCTGGGGCCTCAATGAAGGTCTCCTG
E1L VH-VL-P
CAAGGCTTCTGGGTACACCTTCACCAACTACTATATACACTGGGTGCGACAGGCCCCTGGACAAG
GTCTTGAGTGGATGGGTTGGATCAACCCTAACAGTGGTGCCACAAACTATGCACAGAAGTTCCAG
GGCAGAGTCACCATGACCAGGGACACGTCCACGAGCACAGTCTACATGGAGCTGAGCAGCCTGAG
ATCTGAGGACACGGCCGTGTATTACTGTGCGAAATCCTGGGTCTCCTGGTTTGCTTCCTGGGGTC
AAGGAACCTTGGTCACCGTCTCCTCAGGTGGTGGTGGTTCTGGCGGCGGCGGCTCCGGTGGTGGT
GGTTCTGAGCTCGTGATGACCCAGTCTCCAGACTCCCTGGCTGTGTCTCTGGGCGAGAGGGCCAC
CATCAACTGCAAGTCCAGCCAGAGTGTTTTAAACAGCTCCAACAATAGGAACTACTTAGCTTGGT
ACCAGCAGAAACCAGGACAGCCTCCTAAGCTGCTCATTTACTGGGCATCTACCCGGGAATCCGGG
GTCCCTGACCGATTCAGTGGCAGCGGGTCTGGGACAGATTTCACTCTCACCATCAGTGGCCTGCA
GGCTGAAGATGTGGCAGTTTATTACTGTCAGCAACATTATAGTACTCCATTCACTTTTGGCCCTG
GGACCAAAGTGGATATCAAATCCGGAGGTGGTGGATCCGAGGTGCAGCTGCTCGAGTCTGGAGGA
GGATTGGTGCAGCCTGGAGGGTCATTGAAACTCTCATGTGCAGCCTCTGGATTCACCTTCAATAA
GTACGCCATGAACTGGGTCCGCCAGGCTCCAGGAAAGGGTTTGGAATGGGTTGCTCGCATAAGAA
GTAAATATAATAATTATGCAACATATTATGCCGATTCAGTGAAATCGAGGTTCACCATCTCCAGA
GATGATTCAAAAAACACTGCCTATCTACAAATGAACAACTTGAAAACTGAGGACACTGCCGTGTA
CTACTGTGTGAGACATGGGAACTTCGGTAATAGCTACACATCCTACTACGCTTACTGGGGCCAAG
GGACTCTGGTCACCGTCTCCTCAGGTGGTGGTGGTTCTGGCGGCGGCGGCTCCGGTGGTGGTGGT
TCTGAGCTCGTTGTGACTCAGGAACCTTCACTCACCGTATCACCTGGTGGAACAGTCACACTCAC
TTGTGGCTCCTCGACTGGGGCTGTTACATCTGGCTACTACCCAAACTGGGTCCAACAAAAACCAG
GTCAGGCACCCCGTGGTCTAATAGGTGGGACTAAGTTCCTCGCCCCCGGTACTCCTGCCAGATTC
TCAGGCTCCCTGCTTGGAGGCAAGGCTGCCCTCACCCTCTCAGGGGTACAGCCAGAGGATGAGGC
AGAATATTACTGTGCTCTATGGTACAGCAACCGCTGGGTGTTCGGTGGAGGAACCAAACTGACTG
TCCTA 207. MCSP-G4 VH-VL-P .times. artificial aa
QVQLVQSGAEVKRPGASMKVSCKASGYTFTNYYIHWVRQAPGQGLEWMGWINPNSGATNYAQKFQ
E2M VH-VL-P
GRVTMTRDTSTSTVYMELSSLRSEDTAVYYCAKSWVSWFASWGQGTLVTVSSGGGGSGGGGSGGG
GSELVMTQSPDSLAVSLGERATINCKSSQSVLNSSNNRNYLAWYQQKPGQPPKLLIYWASTRESG
VPDRFSGSGSGTDFTLTISGLQAEDVAVYYCQQHYSTPFTFGPGTKVDIKSGGGGSEVQLLESGG
GLVQPGGSLKLSCAASGFTFNGYAMNWVRQAPGKGLEWVARIRSKYNNYATYYADSVKERFTISR
DDSKNTAYLQMNNLKTEDTAVYYCVRHRNFGNSYLSWFAYWGQGTLVTVSSGGGGSGGGGSGGGG
SELVVTQEPSLTVSPGGTVTLTCRSSTGAVTSGYYPNWVQQKPGQAPRGLIGATDMRPSGTPARF
SGSLLGGKAALTLSGVQPEDEAEYYCALWYSNRWVFGGGTKLTVL 208. MCSP-G4 VH-VL-P
.times. artificial nt
CAGGTGCAGCTGGTCCAGTCTGGGGCTGAGGTGAAGAGGCCTGGGGCCTCAATGAAGGTCTCCTG
E2M VH-VL-P
CAAGGCTTCTGGGTACACCTTCACCAACTACTATATACACTGGGTGCGACAGGCCCCTGGACAAG
GTCTTGAGTGGATGGGTTGGATCAACCCTAACAGTGGTGCCACAAACTATGCACAGAAGTTCCAG
GGCAGAGTCACCATGACCAGGGACACGTCCACGAGCACAGTCTACATGGAGCTGAGCAGCCTGAG
ATCTGAGGACACGGCCGTGTATTACTGTGCGAAATCCTGGGTCTCCTGGTTTGCTTCCTGGGGTC
AAGGAACCTTGGTCACCGTCTCCTCAGGTGGTGGTGGTTCTGGCGGCGGCGGCTCCGGTGGTGGT
GGTTCTGAGCTCGTGATGACCCAGTCTCCAGACTCCCTGGCTGTGTCTCTGGGCGAGAGGGCCAC
CATCAACTGCAAGTCCAGCCAGAGTGTTTTAAACAGCTCCAACAATAGGAACTACTTAGCTTGGT
ACCAGCAGAAACCAGGACAGCCTCCTAAGCTGCTCATTTACTGGGCATCTACCCGGGAATCCGGG
GTCCCTGACCGATTCAGTGGCAGCGGGTCTGGGACAGATTTCACTCTCACCATCAGTGGCCTGCA
GGCTGAAGATGTGGCAGTTTATTACTGTCAGCAACATTATAGTACTCCATTCACTTTTGGCCCTG
GGACCAAAGTGGATATCAAATCCGGAGGTGGTGGATCCGAGGTGCAGCTGCTCGAGTCTGGAGGA
GGATTGGTGCAGCCTGGAGGGTCATTGAAACTCTCATGTGCAGCCTCTGGATTCACCTTCAATGG
CTACGCCATGAACTGGGTCCGCCAGGCTCCAGGAAAGGGTTTGGAATGGGTTGCTCGCATAAGAA
GTAAATATAATAATTATGCAACATATTATGCCGATTCAGTGAAAGAGAGGTTCACCATCTCCAGA
GATGATTCAAAAAACACTGCCTATCTACAAATGAACAACTTGAAAACTGAGGACACTGCCGTGTA
CTACTGTGTGAGACATAGGAACTTCGGTAATAGCTACTTATCCTGGTTCGCTTACTGGGGCCAAG
GGACTCTGGTCACCGTCTCCTCAGGTGGTGGTGGTTCTGGCGGCGGCGGCTCCGGTGGTGGTGGT
TCTGAGCTCGTTGTGACTCAGGAACCTTCACTCACCGTATCACCTGGTGGAACAGTCACACTCAC
TTGTCGCTCCTCGACTGGGGCTGTTACATCTGGCTACTACCCAAACTGGGTCCAACAAAAACCAG
GTCAGGCACCCCGTGGTCTAATAGGTGCCACTGACATGAGGCCCTCTGGTACTCCTGCCAGATTC
TCAGGCTCCCTGCTTGGAGGCAAGGCTGCCCTCACCCTCTCAGGGGTACAGCCAGAGGATGAGGC
AGAATATTACTGTGCTCTATGGTACAGCAACCGCTGGGTGTTCGGTGGAGGAACCAAACTGACTG
TCCTA 209. MCSP-G4 VH-VL-P .times. artificial aa
QVQLVQSGAEVKRPGASMKVSCKASGYTFTNYYIHWVRQAPGQGLEWMGWINPNSGATNYAQKFQ
F7O VH-VL-P
GRVTMTRDTSTSTVYMELSSLRSEDTAVYYCAKSWVSWFASWGQGTLVTVSSGGGGSGGGGSGGG
GSELVMTQSPDSLAVSLGERATINCKSSQSVLNSSNNRNYLAWYQQKPGQPPKLLIYWASTRESG
VPDRFSGSGSGTDFTLTISGLQAEDVAVYYCQQHYSTPFTFGPGTKVDIKSGGGGSEVQLLESGG
GLVQPGGSLKLSCAASGFTFNVYAMNWVRQAPGKGLEWVARIRSKYNNYATYYADSVKKRFTISR
DDSKNTAYLQMNNLKTEDTAVYYCVRHGNFGNSYISWWAYWGQGTLVTVSSGGGGSGGGGSGGGG
SELVVTQEPSLTVSPGGTVTLTCGSSTGAVTSGYYPNWVQQKPGQAPRGLIGGTKFLAPGTPARF
SGSLLGGKAALTLSGVQPEDEAEYYCALWYSNRWVFGGGTKLTVL 210. MCSP-G4 VH-VL-P
.times. artificial nt
CAGGTGCAGCTGGTCCAGTCTGGGGCTGAGGTGAAGAGGCCTGGGGCCTCAATGAAGGTCTCCTG
F7O VH-VL-P
CAAGGCTTCTGGGTACACCTTCACCAACTACTATATACACTGGGTGCGACAGGCCCCTGGACAAG
GTCTTGAGTGGATGGGTTGGATCAACCCTAACAGTGGTGCCACAAACTATGCACAGAAGTTCCAG
GGCAGAGTCACCATGACCAGGGACACGTCCACGAGCACAGTCTACATGGAGCTGAGCAGCCTGAG
ATCTGAGGACACGGCCGTGTATTACTGTGCGAAATCCTGGGTCTCCTGGTTTGCTTCCTGGGGTC
AAGGAACCTTGGTCACCGTCTCCTCAGGTGGTGGTGGTTCTGGCGGCGGCGGCTCCGGTGGTGGT
GGTTCTGAGCTCGTGATGACCCAGTCTCCAGACTCCCTGGCTGTGTCTCTGGGCGAGAGGGCCAC
CATCAACTGCAAGTCCAGCCAGAGTGTTTTAAACAGCTCCAACAATAGGAACTACTTAGCTTGGT
ACCAGCAGAAACCAGGACAGCCTCCTAAGCTGCTCATTTACTGGGCATCTACCCGGGAATCCGGG
GTCCCTGACCGATTCAGTGGCAGCGGGTCTGGGACAGATTTCACTCTCACCATCAGTGGCCTGCA
GGCTGAAGATGTGGCAGTTTATTACTGTCAGCAACATTATAGTACTCCATTCACTTTTGGCCCTG
GGACCAAAGTGGATATCAAATCCGGAGGTGGTGGATCCGAGGTGCAGCTGCTCGAGTCTGGAGGA
GGATTGGTGCAGCCTGGAGGGTCATTGAAACTCTCATGTGCAGCCTCTGGATTCACCTTCAATGT
GTACGCCATGAACTGGGTCCGCCAGGCTCCAGGAAAGGGTTTGGAATGGGTTGCTCGCATAAGAA
GTAAATATAATAATTATGCAACATATTATGCCGATTCAGTGAAAAAGAGGTTCACCATCTCCAGA
GATGATTCAAAAAACACTGCCTATCTACAAATGAACAACTTGAAAACTGAGGACACTGCCGTGTA
CTACTGTGTGAGACATGGGAACTTCGGTAATAGCTACATATCCTGGTGGGCTTACTGGGGCCAAG
GGACTCTGGTCACCGTCTCCTCAGGTGGTGGTGGTTCTGGCGGCGGCGGCTCCGGTGGTGGTGGT
TCTGAGCTCGTTGTGACTCAGGAACCTTCACTCACCGTATCACCTGGTGGAACAGTCACACTCAC
TTGTGGCTCCTCGACTGGGGCTGTTACATCTGGCTACTACCCAAACTGGGTCCAACAAAAACCAG
GTCAGGCACCCCGTGGTCTAATAGGTGGGACTAAGTTCCTCGCCCCCGGTACTCCTGCCAGATTC
TCAGGCTCCCTGCTTGGAGGCAAGGCTGCCCTCACCCTCTCAGGGGTACAGCCAGAGGATGAGGC
AGAATATTACTGTGCTCTATGGTACAGCAACCGCTGGGTGTTCGGTGGAGGAACCAAACTGACTG
TCCTA 211. MCSP-G4 VH-VL-P .times. artificial aa
QVQLVQSGAEVKRPGASMKVSCKASGYTFTNYYIHWVRQAPGQGLEWMGWINPNSGATNYAQKFQ
F12Q VH-VL
GRVTMTRDTSTSTVYMELSSLRSEDTAVYYCAKSWVSWFASWGQGTLVTVSSGGGGSGGGGSGGG
GSELVMTQSPDSLAVSLGERATINCKSSQSVLNSSNNRNYLAWYQQKPGQPPKLLIYWASTRESG
VPDRFSGSGSGTDFTLTISGLQAEDVAVYYCQQHYSTPFTFGPGTKVDIKSGGGGSEVQLVESGG
GLVQPGGSLKLSCAASGFTFNSYAMNWVRQAPGKGLEWVARIRSKYNNYATYYADSVKGRFTISR
DDSKNTAYLQMNNLKTEDTAVYYCVRHGNFGNSYVSWWAYWGQGTLVTVSSGGGGSGGGGSGGGG
SQTVVTQEPSLTVSPGGTVTLTCGSSTGAVTSGNYPNWVQQKPGQAPRGLIGGTKFLAPGTPARF
SGSLLGGKAALTLSGVQPEDEAEYYCVLWYSNRWVFGGGTKLTVL 212. MCSP-G4 VH-VL-P
.times. artificial nt
CAGGTGCAGCTGGTCCAGTCTGGGGCTGAGGTGAAGAGGCCTGGGGCCTCAATGAAGGTCTCCTG
F12Q VH-VL
CAAGGCTTCTGGGTACACCTTCACCAACTACTATATACACTGGGTGCGACAGGCCCCTGGACAAG
GTCTTGAGTGGATGGGTTGGATCAACCCTAACAGTGGTGCCACAAACTATGCACAGAAGTTCCAG
GGCAGAGTCACCATGACCAGGGACACGTCCACGAGCACAGTCTACATGGAGCTGAGCAGCCTGAG
ATCTGAGGACACGGCCGTGTATTACTGTGCGAAATCCTGGGTCTCCTGGTTTGCTTCCTGGGGTC
AAGGAACCTTGGTCACCGTCTCCTCAGGTGGTGGTGGTTCTGGCGGCGGCGGCTCCGGTGGTGGT
GGTTCTGAGCTCGTGATGACCCAGTCTCCAGACTCCCTGGCTGTGTCTCTGGGCGAGAGGGCCAC
CATCAACTGCAAGTCCAGCCAGAGTGTTTTAAACAGCTCCAACAATAGGAACTACTTAGCTTGGT
ACCAGCAGAAACCAGGACAGCCTCCTAAGCTGCTCATTTACTGGGCATCTACCCGGGAATCCGGG
GTCCCTGACCGATTCAGTGGCAGCGGGTCTGGGACAGATTTCACTCTCACCATCAGTGGCCTGCA
GGCTGAAGATGTGGCAGTTTATTACTGTCAGCAACATTATAGTACTCCATTCACTTTTGGCCCTG
GGACCAAAGTGGATATCAAATCCGGAGGTGGTGGATCCGAGGTGCAGCTGGTCGAGTCTGGAGGA
GGATTGGTGCAGCCTGGAGGGTCATTGAAACTCTCATGTGCAGCCTCTGGATTCACCTTCAATAG
CTACGCCATGAACTGGGTCCGCCAGGCTCCAGGAAAGGGTTTGGAATGGGTTGCTCGCATAAGAA
GTAAATATAATAATTATGCAACATATTATGCCGATTCAGTGAAAGGCAGGTTCACCATCTCCAGA
GATGATTCAAAAAACACTGCCTATCTACAAATGAACAACTTGAAAACTGAGGACACTGCCGTGTA
CTACTGTGTGAGACATGGGAACTTCGGTAATAGCTACGTTTCCTGGTGGGCTTACTGGGGCCAAG
GGACTCTGGTCACCGTCTCCTCAGGTGGTGGTGGTTCTGGCGGCGGCGGCTCCGGTGGTGGTGGT
TCTCAGACTGTTGTGACTCAGGAACCTTCACTCACCGTATCACCTGGTGGAACAGTCACACTCAC
TTGTGGCTCCTCGACTGGGGCTGTTACATCTGGCAACTACCCAAACTGGGTCCAACAAAAACCAG
GTCAGGCACCCCGTGGTCTAATAGGTGGGACTAAGTTCCTCGCCCCCGGTACTCCTGCCAGATTC
TCAGGCTCCCTGCTTGGAGGCAAGGCTGCCCTCACCCTCTCAGGGGTACAGCCAGAGGATGAGGC
AGAATATTACTGTGTTCTATGGTACAGCAACCGCTGGGTGTTCGGTGGAGGAACCAAACTGACTG
TCCTA 213. MCSP-G4 VH-VL-P .times. artificial aa
QVQLVQSGAEVKRPGASMKVSCKASGYTFTNYYIHWVRQAPGQGLEWMGWINPNSGATNYAQKFQ
I2C VH-VL
GRVTMTRDTSTSTVYMELSSLRSEDTAVYYCAKSWVSWFASWGQGTLVTVSSGGGGSGGGGSGGG
GSELVMTQSPDSLAVSLGERATINCKSSQSVLNSSNNRNYLAWYQQKPGQPPKLLIYWASTRESG
VPDRFSGSGSGTDFTLTISGLQAEDVAVYYCQQHYSTPFTFGPGTKVDIKSGGGGSEVQLVESGG
GLVQPGGSLKLSCAASGFTFNKYAMNWVRQAPGKGLEWVARIRSKYNNYATYYADSVKDRFTISR
DDSKNTAYLQMNNLKTEDTAVYYCVRHGNFGNSYISYWAYWGQGTLVTVSSGGGGSGGGGSGGGG
SQTVVTQEPSLTVSPGGTVTLTCGSSTGAVTSGNYPNWVQQKPGQAPRGLIGGTKFLAPGTPARF
SGSLLGGKAALTLSGVQPEDEAEYYCVLWYSNRWVFGGGTKLTVL 214. MCSP-G4 VH-VL-P
.times. artificial nt
CAGGTGCAGCTGGTCCAGTCTGGGGCTGAGGTGAAGAGGCCTGGGGCCTCAATGAAGGTCTCCTG
I2C VH-VL
CAAGGCTTCTGGGTACACCTTCACCAACTACTATATACACTGGGTGCGACAGGCCCCTGGACAAG
GTCTTGAGTGGATGGGTTGGATCAACCCTAACAGTGGTGCCACAAACTATGCACAGAAGTTCCAG
GGCAGAGTCACCATGACCAGGGACACGTCCACGAGCACAGTCTACATGGAGCTGAGCAGCCTGAG
ATCTGAGGACACGGCCGTGTATTACTGTGCGAAATCCTGGGTCTCCTGGTTTGCTTCCTGGGGTC
AAGGAACCTTGGTCACCGTCTCCTCAGGTGGTGGTGGTTCTGGCGGCGGCGGCTCCGGTGGTGGT
GGTTCTGAGCTCGTGATGACCCAGTCTCCAGACTCCCTGGCTGTGTCTCTGGGCGAGAGGGCCAC
CATCAACTGCAAGTCCAGCCAGAGTGTTTTAAACAGCTCCAACAATAGGAACTACTTAGCTTGGT
ACCAGCAGAAACCAGGACAGCCTCCTAAGCTGCTCATTTACTGGGCATCTACCCGGGAATCCGGG
GTCCCTGACCGATTCAGTGGCAGCGGGTCTGGGACAGATTTCACTCTCACCATCAGTGGCCTGCA
GGCTGAAGATGTGGCAGTTTATTACTGTCAGCAACATTATAGTACTCCATTCACTTTTGGCCCTG
GGACCAAAGTGGATATCAAATCCGGAGGTGGTGGATCCGAGGTGCAGCTGGTCGAGTCTGGAGGA
GGATTGGTGCAGCCTGGAGGGTCATTGAAACTCTCATGTGCAGCCTCTGGATTCACCTTCAATAA
GTACGCCATGAACTGGGTCCGCCAGGCTCCAGGAAAGGGTTTGGAATGGGTTGCTCGCATAAGAA
GTAAATATAATAATTATGCAACATATTATGCCGATTCAGTGAAAGACAGGTTCACCATCTCCAGA
GATGATTCAAAAAACACTGCCTATCTACAAATGAACAACTTGAAAACTGAGGACACTGCCGTGTA
CTACTGTGTGAGACATGGGAACTTCGGTAATAGCTACATATCCTACTGGGCTTACTGGGGCCAAG
GGACTCTGGTCACCGTCTCCTCAGGTGGTGGTGGTTCTGGCGGCGGCGGCTCCGGTGGTGGTGGT
TCTCAGACTGTTGTGACTCAGGAACCTTCACTCACCGTATCACCTGGTGGAACAGTCACACTCAC
TTGTGGCTCCTCGACTGGGGCTGTTACATCTGGCAACTACCCAAACTGGGTCCAACAAAAACCAG
GTCAGGCACCCCGTGGTCTAATAGGTGGGACTAAGTTCCTCGCCCCCGGTACTCCTGCCAGATTC
TCAGGCTCCCTGCTTGGAGGCAAGGCTGCCCTCACCCTCTCAGGGGTACAGCCAGAGGATGAGGC
AGAATATTACTGTGTTCTATGGTACAGCAACCGCTGGGTGTTCGGTGGAGGAACCAAACTGACTG
TCCTA 215. MCSP-D2 VH-VL .times. artificial aa
QVQLVQSGAEVKKPGASVKVSCKASGYTFTGYYMHWVRQAPGQGLEWMGWINPNSGGTSYAQKFQ
H2C VH-VL
GRVTMTRDTSTSTVYMELSNLRSDDTAVYYCAKSWVSWFASWGQGTLVTVSSGGGGSGGGGSGGG
GSDIVMTQSPDSLAVSLGERATINCKSSQSVLNSSNNRNYLAWYQQKPGQPPKLLIYWASTRESG
VPDRFSGSGSGTDFTLTISGLQAEDVAVYYCQQHYSTPFTFGPGTKVDIKSGGGGSEVQLVESGG
GLVQPGGSLKLSCAASGFTFNKYAMNWVRQAPGKGLEWVARIRSKYNNYATYYADSVKDRFTISR
DDSKNTAYLQMNNLKTEDTAVYYCVRHGNFGNSYISYWAYWGQGTLVTVSSGGGGSGGGGSGGGG
SQTVVTQEPSLTVSPGGTVTLTCGSSTGAVTSGYYPNWVQQKPGQAPRGLIGGTKFLAPGTPARF
SGSLLGGKAALTLSGVQPEDEAEYYCALWYSNRWVFGGGTKLTVL 216. MCSP-D2 VH-VL
.times. artificial nt
CAGGTGCAGCTGGTCCAGTCTGGGGCTGAGGTGAAGAAGCCTGGGGCCTCAGTGAAGGTCTCCTG
H2C VH-VL
CAAGGCTTCTGGATACACCTTCACCGGCTACTATATGCACTGGGTGCGACAGGCCCCTGGACAAG
GGCTTGAGTGGATGGGATGGATCAACCCTAACAGTGGTGGCACAAGCTACGCACAGAAGTTCCAG
GGCAGAGTCACCATGACTAGGGACACGTCCACGAGCACAGTCTACATGGAGCTGAGCAACCTGAG
ATCTGACGACACGGCCGTGTATTACTGTGCGAAATCCTGGGTCTCCTGGTTTGCTTCCTGGGGTC
AAGGAACCTTGGTCACCGTCTCCTCAGGTGGTGGTGGTTCTGGCGGCGGCGGCTCCGGTGGTGGT
GGTTCTGATATCGTGATGACCCAGTCTCCAGACTCCCTGGCTGTGTCTCTGGGCGAGAGGGCCAC
CATCAACTGCAAGTCCAGCCAGAGTGTTTTAAACAGCTCCAACAATAGGAACTACTTAGCTTGGT
ACCAGCAGAAACCAGGACAGCCTCCTAAGCTGCTCATTTACTGGGCATCTACCCGGGAATCCGGG
GTCCCTGACCGATTCAGTGGCAGCGGGTCTGGGACAGATTTCACTCTCACCATCAGTGGCCTGCA
GGCTGAAGATGTGGCAGTTTATTACTGTCAGCAACATTATAGTACTCCATTCACTTTTGGCCCTG
GGACCAAAGTGGATATCAAATCCGGAGGTGGTGGATCCGAGGTGCAGCTGGTCGAGTCTGGAGGA
GGATTGGTGCAGCCTGGAGGGTCATTGAAACTCTCATGTGCAGCCTCTGGATTCACCTTCAATAA
GTACGCCATGAACTGGGTCCGCCAGGCTCCAGGAAAGGGTTTGGAATGGGTTGCTCGCATAAGAA
GTAAATATAATAATTATGCAACATATTATGCCGATTCAGTGAAAGACAGGTTCACCATCTCCAGA
GATGATTCAAAAAACACTGCCTATCTACAAATGAACAACTTGAAAACTGAGGACACTGCCGTGTA
CTACTGTGTGAGACATGGGAACTTCGGTAATAGCTACATATCCTACTGGGCTTACTGGGGCCAAG
GGACTCTGGTCACCGTCTCCTCAGGTGGTGGTGGTTCTGGCGGCGGCGGCTCCGGTGGTGGTGGT
TCTCAGACTGTTGTGACTCAGGAACCTTCACTCACCGTATCACCTGGTGGAACAGTCACACTCAC
TTGTGGCTCCTCGACTGGGGCTGTTACATCTGGCTACTACCCAAACTGGGTCCAACAAAAACCAG
GTCAGGCACCCCGTGGTCTAATAGGTGGGACTAAGTTCCTCGCCCCCGGTACTCCTGCCAGATTC
TCAGGCTCCCTGCTTGGAGGCAAGGCTGCCCTCACCCTCTCAGGGGTACAGCCAGAGGATGAGGC
AGAATATTACTGTGCTCTATGGTACAGCAACCGCTGGGTGTTCGGTGGAGGAACCAAACTGACTG
TCCTA 217. MCSP-D2 VH-VL .times. artificial aa
QVQLVQSGAEVKKPGASVKVSCKASGYTFTGYYMHWVRQAPGQGLEWMGWINPNSGGTSYAQKFQ
F12Q VH-VL
GRVTMTRDTSTSTVYMELSNLRSDDTAVYYCAKSWVSWFASWGQGTLVTVSSGGGGSGGGGSGGG
GSDIVMTQSPDSLAVSLGERATINCKSSQSVLNSSNNRNYLAWYQQKPGQPPKLLIYWASTRESG
VPDRFSGSGSGTDFTLTISGLQAEDVAVYYCQQHYSTPFTFGPGTKVDIKSGGGGSEVQLVESGG
GLVQPGGSLKLSCAASGFTFNSYAMNWVRQAPGKGLEWVARIRSKYNNYATYYADSVKGRFTISR
DDSKNTAYLQMNNLKTEDTAVYYCVRHGNFGNSYVSWWAYWGQGTLVTVSSGGGGSGGGGSGGGG
SQTVVTQEPSLTVSPGGTVTLTCGSSTGAVTSGNYPNWVQQKPGQAPRGLIGGTKFLAPGTPARF
SGSLLGGKAALTLSGVQPEDEAEYYCVLWYSNRWVFGGGTKLTVL 218. MCSP-D2 VH-VL
.times. artificial nt
CAGGTGCAGCTGGTCCAGTCTGGGGCTGAGGTGAAGAAGCCTGGGGCCTCAGTGAAGGTCTCCTG
F12Q VH-VL
CAAGGCTTCTGGATACACCTTCACCGGCTACTATATGCACTGGGTGCGACAGGCCCCTGGACAAG
GGCTTGAGTGGATGGGATGGATCAACCCTAACAGTGGTGGCACAAGCTACGCACAGAAGTTCCAG
GGCAGAGTCACCATGACTAGGGACACGTCCACGAGCACAGTCTACATGGAGCTGAGCAACCTGAG
ATCTGACGACACGGCCGTGTATTACTGTGCGAAATCCTGGGTCTCCTGGTTTGCTTCCTGGGGTC
AAGGAACCTTGGTCACCGTCTCCTCAGGTGGTGGTGGTTCTGGCGGCGGCGGCTCCGGTGGTGGT
GGTTCTGATATCGTGATGACCCAGTCTCCAGACTCCCTGGCTGTGTCTCTGGGCGAGAGGGCCAC
CATCAACTGCAAGTCCAGCCAGAGTGTTTTAAACAGCTCCAACAATAGGAACTACTTAGCTTGGT
ACCAGCAGAAACCAGGACAGCCTCCTAAGCTGCTCATTTACTGGGCATCTACCCGGGAATCCGGG
GTCCCTGACCGATTCAGTGGCAGCGGGTCTGGGACAGATTTCACTCTCACCATCAGTGGCCTGCA
GGCTGAAGATGTGGCAGTTTATTACTGTCAGCAACATTATAGTACTCCATTCACTTTTGGCCCTG
GGACCAAAGTGGATATCAAATCCGGAGGTGGTGGATCCGAGGTGCAGCTGGTCGAGTCTGGAGGA
GGATTGGTGCAGCCTGGAGGGTCATTGAAACTCTCATGTGCAGCCTCTGGATTCACCTTCAATAG
CTACGCCATGAACTGGGTCCGCCAGGCTCCAGGAAAGGGTTTGGAATGGGTTGCTCGCATAAGAA
GTAAATATAATAATTATGCAACATATTATGCCGATTCAGTGAAAGGCAGGTTCACCATCTCCAGA
GATGATTCAAAAAACACTGCCTATCTACAAATGAACAACTTGAAAACTGAGGACACTGCCGTGTA
CTACTGTGTGAGACATGGGAACTTCGGTAATAGCTACGTTTCCTGGTGGGCTTACTGGGGCCAAG
GGACTCTGGTCACCGTCTCCTCAGGTGGTGGTGGTTCTGGCGGCGGCGGCTCCGGTGGTGGTGGT
TCTCAGACTGTTGTGACTCAGGAACCTTCACTCACCGTATCACCTGGTGGAACAGTCACACTCAC
TTGTGGCTCCTCGACTGGGGCTGTTACATCTGGCAACTACCCAAACTGGGTCCAACAAAAACCAG
GTCAGGCACCCCGTGGTCTAATAGGTGGGACTAAGTTCCTCGCCCCCGGTACTCCTGCCAGATTC
TCAGGCTCCCTGCTTGGAGGCAAGGCTGCCCTCACCCTCTCAGGGGTACAGCCAGAGGATGAGGC
AGAATATTACTGTGTTCTATGGTACAGCAACCGCTGGGTGTTCGGTGGAGGAACCAAACTGACTG
TCCTA 219. MCSP-D2 VH-VL .times. artificial aa
QVQLVQSGAEVKKPGASVKVSCKASGYTFTGYYMHWVRQAPGQGLEWMGWINPNSGGTSYAQKFQ
I2C VH-VL
GRVTMTRDTSTSTVYMELSNLRSDDTAVYYCAKSWVSWFASWGQGTLVTVSSGGGGSGGGGSGGG
GSDIVMTQSPDSLAVSLGERATINCKSSQSVLNSSNNRNYLAWYQQKPGQPPKLLIYWASTRESG
VPDRFSGSGSGTDFTLTISGLQAEDVAVYYCQQHYSTPFTFGPGTKVDIKSGGGGSEVQLVESGG
GLVQPGGSLKLSCAASGFTFNKYAMNWVRQAPGKGLEWVARIRSKYNNYATYYADSVKDRFTISR
DDSKNTAYLQMNNLKTEDTAVYYCVRHGNFGNSYISYWAYWGQGTLVTVSSGGGGSGGGGSGGGG
SQTVVTQEPSLTVSPGGTVTLTCGSSTGAVTSGNYPNWVQQKPGQAPRGLIGGTKFLAPGTPARF
SGSLLGGKAALTLSGVQPEDEAEYYCVLWYSNRWVFGGGTKLTVL 220. MCSP-D2 VH-VL
.times. artificial nt
CAGGTGCAGCTGGTCCAGTCTGGGGCTGAGGTGAAGAAGCCTGGGGCCTCAGTGAAGGTCTCCTG
I2C VH-VL
CAAGGCTTCTGGATACACCTTCACCGGCTACTATATGCACTGGGTGCGACAGGCCCCTGGACAAG
GGCTTGAGTGGATGGGATGGATCAACCCTAACAGTGGTGGCACAAGCTACGCACAGAAGTTCCAG
GGCAGAGTCACCATGACTAGGGACACGTCCACGAGCACAGTCTACATGGAGCTGAGCAACCTGAG
ATCTGACGACACGGCCGTGTATTACTGTGCGAAATCCTGGGTCTCCTGGTTTGCTTCCTGGGGTC
AAGGAACCTTGGTCACCGTCTCCTCAGGTGGTGGTGGTTCTGGCGGCGGCGGCTCCGGTGGTGGT
GGTTCTGATATCGTGATGACCCAGTCTCCAGACTCCCTGGCTGTGTCTCTGGGCGAGAGGGCCAC
CATCAACTGCAAGTCCAGCCAGAGTGTTTTAAACAGCTCCAACAATAGGAACTACTTAGCTTGGT
ACCAGCAGAAACCAGGACAGCCTCCTAAGCTGCTCATTTACTGGGCATCTACCCGGGAATCCGGG
GTCCCTGACCGATTCAGTGGCAGCGGGTCTGGGACAGATTTCACTCTCACCATCAGTGGCCTGCA
GGCTGAAGATGTGGCAGTTTATTACTGTCAGCAACATTATAGTACTCCATTCACTTTTGGCCCTG
GGACCAAAGTGGATATCAAATCCGGAGGTGGTGGATCCGAGGTGCAGCTGGTCGAGTCTGGAGGA
GGATTGGTGCAGCCTGGAGGGTCATTGAAACTCTCATGTGCAGCCTCTGGATTCACCTTCAATAA
GTACGCCATGAACTGGGTCCGCCAGGCTCCAGGAAAGGGTTTGGAATGGGTTGCTCGCATAAGAA
GTAAATATAATAATTATGCAACATATTATGCCGATTCAGTGAAAGACAGGTTCACCATCTCCAGA
GATGATTCAAAAAACACTGCCTATCTACAAATGAACAACTTGAAAACTGAGGACACTGCCGTGTA
CTACTGTGTGAGACATGGGAACTTCGGTAATAGCTACATATCCTACTGGGCTTACTGGGGCCAAG
GGACTCTGGTCACCGTCTCCTCAGGTGGTGGTGGTTCTGGCGGCGGCGGCTCCGGTGGTGGTGGT
TCTCAGACTGTTGTGACTCAGGAACCTTCACTCACCGTATCACCTGGTGGAACAGTCACACTCAC
TTGTGGCTCCTCGACTGGGGCTGTTACATCTGGCAACTACCCAAACTGGGTCCAACAAAAACCAG
GTCAGGCACCCCGTGGTCTAATAGGTGGGACTAAGTTCCTCGCCCCCGGTACTCCTGCCAGATTC
TCAGGCTCCCTGCTTGGAGGCAAGGCTGCCCTCACCCTCTCAGGGGTACAGCCAGAGGATGAGGC
AGAATATTACTGTGTTCTATGGTACAGCAACCGCTGGGTGTTCGGTGGAGGAACCAAACTGACTG
TCCTA 221. MCSP-D2 VH-VL-P .times. artificial aa
QVQLVQSGAEVKKPGASVKVSCKASGYTFTGYYMHWVRQAPGQGLEWMGWINPNSGGTSYAQKFQ
H2C VH-VL-P
GRVTMTRDTSTSTVYMELSNLRSDDTAVYYCAKSWVSWFASWGQGTLVTVSSGGGGSGGGGSGGG
GSELVMTQSPDSLAVSLGERATINCKSSQSVLNSSNNRNYLAWYQQKPGQPPKLLIYWASTRESG
VPDRFSGSGSGTDFTLTISGLQAEDVAVYYCQQHYSTPFTFGPGTKVDIKSGGGGSEVQLLESGG
GLVQPGGSLKLSCAASGFTFNKYAMNWVRQAPGKGLEWVARIRSKYNNYATYYADSVKDRFTISR
DDSKNTAYLQMNNLKTEDTAVYYCVRHGNFGNSYISYWAYWGQGTLVTVSSGGGGSGGGGSGGGG
SELVVTQEPSLTVSPGGTVTLTCGSSTGAVTSGYYPNWVQQKPGQAPRGLIGGTKFLAPGTPARF
SGSLLGGKAALTLSGVQPEDEAEYYCALWYSNRWVFGGGTKLTVL 222. MCSP-D2 VH-VL-P
.times. artificial nt
CAGGTGCAGCTGGTCCAGTCTGGGGCTGAGGTGAAGAAGCCTGGGGCCTCAGTGAAGGTCTCCTG
H2C VH-VL-P
CAAGGCTTCTGGATACACCTTCACCGGCTACTATATGCACTGGGTGCGACAGGCCCCTGGACAAG
GGCTTGAGTGGATGGGATGGATCAACCCTAACAGTGGTGGCACAAGCTACGCACAGAAGTTCCAG
GGCAGAGTCACCATGACTAGGGACACGTCCACGAGCACAGTCTACATGGAGCTGAGCAACCTGAG
ATCTGACGACACGGCCGTGTATTACTGTGCGAAATCCTGGGTCTCCTGGTTTGCTTCCTGGGGTC
AAGGAACCTTGGTCACCGTCTCCTCAGGTGGTGGTGGTTCTGGCGGCGGCGGCTCCGGTGGTGGT
GGTTCTGAGCTCGTGATGACCCAGTCTCCAGACTCCCTGGCTGTGTCTCTGGGCGAGAGGGCCAC
CATCAACTGCAAGTCCAGCCAGAGTGTTTTAAACAGCTCCAACAATAGGAACTACTTAGCTTGGT
ACCAGCAGAAACCAGGACAGCCTCCTAAGCTGCTCATTTACTGGGCATCTACCCGGGAATCCGGG
GTCCCTGACCGATTCAGTGGCAGCGGGTCTGGGACAGATTTCACTCTCACCATCAGTGGCCTGCA
GGCTGAAGATGTGGCAGTTTATTACTGTCAGCAACATTATAGTACTCCATTCACTTTTGGCCCTG
GGACCAAAGTGGATATCAAATCCGGAGGTGGTGGATCCGAGGTGCAGCTGCTCGAGTCTGGAGGA
GGATTGGTGCAGCCTGGAGGGTCATTGAAACTCTCATGTGCAGCCTCTGGATTCACCTTCAATAA
GTACGCCATGAACTGGGTCCGCCAGGCTCCAGGAAAGGGTTTGGAATGGGTTGCTCGCATAAGAA
GTAAATATAATAATTATGCAACATATTATGCCGATTCAGTGAAAGACAGGTTCACCATCTCCAGA
GATGATTCAAAAAACACTGCCTATCTACAAATGAACAACTTGAAAACTGAGGACACTGCCGTGTA
CTACTGTGTGAGACATGGGAACTTCGGTAATAGCTACATATCCTACTGGGCTTACTGGGGCCAAG
GGACTCTGGTCACCGTCTCCTCAGGTGGTGGTGGTTCTGGCGGCGGCGGCTCCGGTGGTGGTGGT
TCTGAGCTCGTTGTGACTCAGGAACCTTCACTCACCGTATCACCTGGTGGAACAGTCACACTCAC
TTGTGGCTCCTCGACTGGGGCTGTTACATCTGGCTACTACCCAAACTGGGTCCAACAAAAACCAG
GTCAGGCACCCCGTGGTCTAATAGGTGGGACTAAGTTCCTCGCCCCCGGTACTCCTGCCAGATTC
TCAGGCTCCCTGCTTGGAGGCAAGGCTGCCCTCACCCTCTCAGGGGTACAGCCAGAGGATGAGGC
AGAATATTACTGTGCTCTATGGTACAGCAACCGCTGGGTGTTCGGTGGAGGAACCAAACTGACTG
TCCTA 223. MCSP-F9 VH-VL .times. artificial aa
QVQLQESGPGLVKPSETLSLTCVVSGGSISSSNWWSWVRQPPGKGLEWLGTIYYNGNTYYNPSLK
H2C VH-VL
SRVTISVDTSKNQFSLRLSSVTAADTAVYYCAKSWVSWFASWGQGTLVTVSSGGGGSGGGGSGGG
GSDIVMTQSPDSLAVSLGERATINCKSSQSVLSSSNNKNYLNWYQQKPGQPPKLLIYWASTRESG
VPDRFSGSGSGTDFTLTISSLQAEDVAVYYCQQHYSVPFTFGPGTKVDIKGGGGSEVQLVESGGG
LVQPGGSLKLSCAASGFTFNKYAMNWVRQAPGKGLEWVARIRSKYNNYATYYADSVKDRFTISRD
DSKNTAYLQMNNLKTEDTAVYYCVRHGNFGNSYISYWAYWGQGTLVTVSSGGGGSGGGGSGGGGS
QTVVTQEPSLTVSPGGTVTLTCGSSTGAVTSGYYPNWVQQKPGQAPRGLIGGTKFLAPGTPARFS
GSLLGGKAALTLSGVQPEDEAEYYCALWYSNRWVFGGGTKLTVL 224. MCSP-F9 VH-VL
.times. artificial nt
CAGGTGCAGCTGCAAGAGTCTGGCCCAGGACTGGTGAAGCCTTCGGAGACCCTGTCCCTCACCTG
H2C VH-VL
CGTTGTCTCTGGTGGCTCCATCAGCAGTAGTAACTGGTGGAGCTGGGTCCGCCAGCCCCCAGGGA
AGGGACTGGAGTGGCTTGGGACTATCTATTATAATGGGAATACCTACTACAACCCGTCCCTCAAG
AGTCGAGTCACCATCTCCGTAGACACGTCCAAGAACCAGTTCTCCCTGAGGCTGAGCTCTGTGAC
CGCCGCAGACACGGCCGTGTATTACTGTGCGAAATCCTGGGTCTCCTGGTTTGCTTCCTGGGGTC
AAGGAACCTTGGTCACCGTCTCCTCAGGTGGTGGTGGTTCTGGCGGCGGCGGCTCCGGTGGTGGT
GGTTCTGATATCGTGATGACACAGTCTCCAGACTCCCTGGCTGTGTCTCTGGGCGAGAGGGCCAC
CATCAACTGCAAGTCCAGCCAGAGTGTCTTATCCAGCTCCAACAATAAGAACTACTTAAATTGGT
ACCAGCAGAAACCAGGACAGCCTCCTAAGTTGCTCATTTACTGGGCATCTACCCGGGAATCCGGG
GTCCCTGACCGATTCAGTGGCAGCGGGTCTGGGACAGATTTCACTCTCACCATCAGCAGCCTGCA
GGCTGAAGATGTGGCAGTTTATTACTGTCAGCAACATTATAGTGTTCCATTCACTTTCGGCCCTG
GGACCAAAGTGGATATCAAAGGAGGTGGTGGCTCCGAGGTGCAGCTGGTCGAGTCTGGAGGAGGA
TTGGTGCAGCCTGGAGGGTCATTGAAACTCTCATGTGCAGCCTCTGGATTCACCTTCAATAAGTA
CGCCATGAACTGGGTCCGCCAGGCTCCAGGAAAGGGTTTGGAATGGGTTGCTCGCATAAGAAGTA
AATATAATAATTATGCAACATATTATGCCGATTCAGTGAAAGACAGGTTCACCATCTCCAGAGAT
GATTCAAAAAACACTGCCTATCTACAAATGAACAACTTGAAAACTGAGGACACTGCCGTGTACTA
CTGTGTGAGACATGGGAACTTCGGTAATAGCTACATATCCTACTGGGCTTACTGGGGCCAAGGGA
CTCTGGTCACCGTCTCCTCAGGTGGTGGTGGTTCTGGCGGCGGCGGCTCCGGTGGTGGTGGTTCT
CAGACTGTTGTGACTCAGGAACCTTCACTCACCGTATCACCTGGTGGAACAGTCACACTCACTTG
TGGCTCCTCGACTGGGGCTGTTACATCTGGCTACTACCCAAACTGGGTCCAACAAAAACCAGGTC
AGGCACCCCGTGGTCTAATAGGTGGGACTAAGTTCCTCGCCCCCGGTACTCCTGCCAGATTCTCA
GGCTCCCTGCTTGGAGGCAAGGCTGCCCTCACCCTCTCAGGGGTACAGCCAGAGGATGAGGCAGA
ATATTACTGTGCTCTATGGTACAGCAACCGCTGGGTGTTCGGTGGAGGAACCAAACTGACTGTCC
TA 225. MCSP-F9 VH-VL-P .times. artificial aa
EVQLQESGPGLVKPSETLSLTCVVSGGSISSSNWWSWVRQPPGKGLEWLGTIYYNGNTYYNPSLK
H2C VH-VL-P
SRVTISVDTSKNQFSLRLSSVTAADTAVYYCAKSWVSWFASWGQGTLVTVSSGGGGSGGGGSGGG
GSELVMTQSPDSLAVSLGERATINCKSSQSVLSSSNNKNYLNWYQQKPGQPPKLLIYWASTRESG
VPDRFSGSGSGTDFTLTISSLQAEDVAVYYCQQHYSVPFTFGPGTKVDIKSGGGGSEVQLLESGG
GLVQPGGSLKLSCAASGFTFNKYAMNWVRQAPGKGLEWVARIRSKYNNYATYYADSVKDRFTISR
DDSKNTAYLQMNNLKTEDTAVYYCVRHGNFGNSYISYWAYWGQGTLVTVSSGGGGSGGGGSGGGG
SELVVTQEPSLTVSPGGTVTLTCGSSTGAVTSGYYPNWVQQKPGQAPRGLIGGTKFLAPGTPARF
SGSLLGGKAALTLSGVQPEDEAEYYCALWYSNRWVFGGGTKLTVL 226. MCSP-F9 VH-VL-P
.times. artificial nt
GAGGTGCAGCTGCAAGAGTCTGGCCCAGGACTGGTGAAGCCTTCGGAGACCCTGTCCCTCACCTG
H2C VH-VL-P
CGTTGTCTCTGGTGGCTCCATCAGCAGTAGTAACTGGTGGAGCTGGGTCCGCCAGCCCCCAGGGA
AGGGACTGGAGTGGCTTGGGACTATCTATTATAATGGGAATACCTACTACAACCCGTCCCTCAAG
AGTCGAGTCACCATCTCCGTAGACACGTCCAAGAACCAGTTCTCCCTGAGGCTGAGCTCTGTGAC
CGCCGCAGACACGGCCGTGTATTACTGTGCGAAATCCTGGGTCTCCTGGTTTGCTTCCTGGGGTC
AAGGAACCTTGGTCACCGTCTCCTCAGGTGGTGGTGGTTCTGGCGGCGGCGGCTCCGGTGGTGGT
GGTTCTGAGCTCGTGATGACACAGTCTCCAGACTCCCTGGCTGTGTCTCTGGGCGAGAGGGCCAC
CATCAACTGCAAGTCCAGCCAGAGTGTCTTATCCAGCTCCAACAATAAGAACTACTTAAATTGGT
ACCAGCAGAAACCAGGACAGCCTCCTAAGTTGCTCATTTACTGGGCATCTACCCGGGAATCCGGG
GTCCCTGACCGATTCAGTGGCAGCGGGTCTGGGACAGATTTCACTCTCACCATCAGCAGCCTGCA
GGCTGAAGATGTGGCAGTTTATTACTGTCAGCAACATTATAGTGTTCCATTCACTTTCGGCCCTG
GGACCAAAGTGGATATCAAATCCGGAGGTGGTGGATCCGAGGTGCAGCTGCTCGAGTCTGGAGGA
GGATTGGTGCAGCCTGGAGGGTCATTGAAACTCTCATGTGCAGCCTCTGGATTCACCTTCAATAA
GTACGCCATGAACTGGGTCCGCCAGGCTCCAGGAAAGGGTTTGGAATGGGTTGCTCGCATAAGAA
GTAAATATAATAATTATGCAACATATTATGCCGATTCAGTGAAAGACAGGTTCACCATCTCCAGA
GATGATTCAAAAAACACTGCCTATCTACAAATGAACAACTTGAAAACTGAGGACACTGCCGTGTA
CTACTGTGTGAGACATGGGAACTTCGGTAATAGCTACATATCCTACTGGGCTTACTGGGGCCAAG
GGACTCTGGTCACCGTCTCCTCAGGTGGTGGTGGTTCTGGCGGCGGCGGCTCCGGTGGTGGTGGT
TCTGAGCTCGTTGTGACTCAGGAACCTTCACTCACCGTATCACCTGGTGGAACAGTCACACTCAC
TTGTGGCTCCTCGACTGGGGCTGTTACATCTGGCTACTACCCAAACTGGGTCCAACAAAAACCAG
GTCAGGCACCCCGTGGTCTAATAGGTGGGACTAAGTTCCTCGCCCCCGGTACTCCTGCCAGATTC
TCAGGCTCCCTGCTTGGAGGCAAGGCTGCCCTCACCCTCTCAGGGGTACAGCCAGAGGATGAGGC
AGAATATTACTGTGCTCTATGGTACAGCAACCGCTGGGTGTTCGGTGGAGGAACCAAACTGACTG
TCCTA
227. MCSP-F9 VH-VL-P .times. artificial aa
EVQLQESGPGLVKPSETLSLTCVVSGGSISSSNWWSWVRQPPGKGLEWLGTIYYNGNTYYNPSLK
G4H VH-VL-P
SRVTISVDTSKNQFSLRLSSVTAADTAVYYCAKSWVSWFASWGQGTLVTVSSGGGGSGGGGSGGG
GSELVMTQSPDSLAVSLGERATINCKSSQSVLSSSNNKNYLNWYQQKPGQPPKLLIYWASTRESG
VPDRFSGSGSGTDFTLTISSLQAEDVAVYYCQQHYSVPFTFGPGTKVDIKSGGGGSEVQLLESGG
GLVQPGGSLKLSCAASGFTFNRYAMNWVRQAPGKGLEWVARIRSKYNNYATYYADSVKGRFTISR
DDSKNTAYLQMNNLKTEDTAVYYCVRHGNFGNSYLSYFAYWGQGTLVTVSSGGGGSGGGGSGGGG
SELVVTQEPSLTVSPGGTVTLTCGSSTGAVTSGYYPNWVQQKPGQAPRGLIGGTKFLAPGTPARF
SGSLLGGKAALTLSGVQPEDEAEYYCALWYSNRWVFGGGTKLTVL 228. MCSP-F9 VH-VL-P
.times. artificial nt
GAGGTGCAGCTGCAAGAGTCTGGCCCAGGACTGGTGAAGCCTTCGGAGACCCTGTCCCTCACCTG
G4H VH-VL-P
CGTTGTCTCTGGTGGCTCCATCAGCAGTAGTAACTGGTGGAGCTGGGTCCGCCAGCCCCCAGGGA
AGGGACTGGAGTGGCTTGGGACTATCTATTATAATGGGAATACCTACTACAACCCGTCCCTCAAG
AGTCGAGTCACCATCTCCGTAGACACGTCCAAGAACCAGTTCTCCCTGAGGCTGAGCTCTGTGAC
CGCCGCAGACACGGCCGTGTATTACTGTGCGAAATCCTGGGTCTCCTGGTTTGCTTCCTGGGGTC
AAGGAACCTTGGTCACCGTCTCCTCAGGTGGTGGTGGTTCTGGCGGCGGCGGCTCCGGTGGTGGT
GGTTCTGAGCTCGTGATGACACAGTCTCCAGACTCCCTGGCTGTGTCTCTGGGCGAGAGGGCCAC
CATCAACTGCAAGTCCAGCCAGAGTGTCTTATCCAGCTCCAACAATAAGAACTACTTAAATTGGT
ACCAGCAGAAACCAGGACAGCCTCCTAAGTTGCTCATTTACTGGGCATCTACCCGGGAATCCGGG
GTCCCTGACCGATTCAGTGGCAGCGGGTCTGGGACAGATTTCACTCTCACCATCAGCAGCCTGCA
GGCTGAAGATGTGGCAGTTTATTACTGTCAGCAACATTATAGTGTTCCATTCACTTTCGGCCCTG
GGACCAAAGTGGATATCAAATCCGGAGGTGGTGGATCCGAGGTGCAGCTGCTCGAGTCTGGAGGA
GGATTGGTGCAGCCTGGAGGGTCATTGAAACTCTCATGTGCAGCCTCTGGATTCACCTTCAATCG
CTACGCCATGAACTGGGTCCGCCAGGCTCCAGGAAAGGGTTTGGAATGGGTTGCTCGCATAAGAA
GTAAATATAATAATTATGCAACATATTATGCCGATTCAGTGAAAGGGAGGTTCACCATCTCCAGA
GATGATTCAAAAAACACTGCCTATCTACAAATGAACAACTTGAAAACTGAGGACACTGCCGTGTA
CTACTGTGTGAGACATGGGAACTTCGGTAATAGCTACTTATCCTACTTCGCTTACTGGGGCCAAG
GGACTCTGGTCACCGTCTCCTCAGGTGGTGGTGGTTCTGGCGGCGGCGGCTCCGGTGGTGGTGGT
TCTGAGCTCGTTGTGACTCAGGAACCTTCACTCACCGTATCACCTGGTGGAACAGTCACACTCAC
TTGTGGCTCCTCGACTGGGGCTGTTACATCTGGCTACTACCCAAACTGGGTCCAACAAAAACCAG
GTCAGGCACCCCGTGGTCTAATAGGTGGGACTAAGTTCCTCGCCCCCGGTACTCCTGCCAGATTC
TCAGGCTCCCTGCTTGGAGGCAAGGCTGCCCTCACCCTCTCAGGGGTACAGCCAGAGGATGAGGC
AGAATATTACTGTGCTCTATGGTACAGCAACCGCTGGGTGTTCGGTGGAGGAACCAAACTGACTG
TCCTA 229. 1-27 CD3.epsilon.-Fc artificial aa
QDGNEEMGGITQTPYKVSISGTTVILTSGEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTL
MISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNG
KEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEW
ESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG
KHHHHHH 230. 1-27 CD3.epsilon.-Fc artificial nt
ATGGGATGGAGCTGTATCATCCTCTTCTTGGTAGCAACAGCTACAGGTGTACACTCCCAAGATGG
TAATGAAGAAATGGGTGGTATTACACAGACACCATATAAAGTCTCCATCTCTGGAACCACAGTAA
TATTGACATCCGGAGAGCCCAAATCTTGTGACAAAACTCACACATGCCCACCGTGCCCAGCACCT
GAACTCCTGGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTC
CCGGACCCCTGAGGTCACATGCGTGGTGGTGGACGTGAGCCACGAAGACCCTGAGGTCAAGTTCA
ACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAAC
AGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTA
CAAGTGCAAGGTCTCCAACAAAGCCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAG
GGCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGAGGAGATGACCAAGAACCAG
GTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAA
TGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCC
TCTATAGCAAGCTCACCGTGGACAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTG
ATGCATGAGGCTCTGCACAACCACTACACGCAGAAGAGCCTCTCCCTGTCCCCGGGTAAACATCA
TCACCATCATCAT 231. human 1-27 CD3.epsilon.- artificial aa
QDGNEEMGGITQTPYKVSISGTTVILTDYKDDDDKTASFAAAQKECVCENYKLAVNCFLNDNGQC
EpCAM
QCTSIGAQNTVLCSKLAAKCLVMKAEMNGSKLGRRAKPEGALQNNDGLYDPDCDESGLFKAKQCN
GTSTCWCVNTAGVRRTDKDTEITCSERVRTYWIIIELKHKAREKPYDVQSLRTALEEAIKTRYQL
DPKFITNILYEDNVITIDLVQNSSQKTQNDVDIADVAYYFEKDVKGESLFHSKKMDLRVNGEQLD
LDPGQTLIYYVDEKAPEFSMQGLKAGVIAVIVVVVIAIVAGIVVLVISRKKRMAKYEKAEIKEMG
EMHRELNA 232. human 1-27 CD3.epsilon.- artificial nt
ATGGGATGGAGCTGTATCATCCTCTTCTTGGTAGCAACAGCTACAGGTGTACACTCCCAAGATGG
EpCAM
TAATGAAGAAATGGGTGGTATTACACAGACACCATATAAAGTCTCCATCTCTGGAACCACAGTAA
TATTGACAGATTACAAGGACGACGATGACAAGACTGCGAGTTTTGCCGCAGCTCAGAAAGAATGT
GTCTGTGAAAACTACAAGCTGGCCGTAAACTGCTTTTTGAATGACAATGGTCAATGCCAGTGTAC
TTCGATTGGTGCACAAAATACTGTCCTTTGCTCAAAGCTGGCTGCCAAATGTTTGGTGATGAAGG
CAGAAATGAACGGCTCAAAACTTGGGAGAAGAGCGAAACCTGAAGGGGCTCTCCAGAACAATGAT
GGCCTTTACGATCCTGACTGCGATGAGAGCGGGCTCTTTAAGGCCAAGCAGTGCAACGGCACCTC
CACGTGCTGGTGTGTGAACACTGCTGGGGTCAGAAGAACTGACAAGGACACTGAAATAACCTGCT
CTGAGCGAGTGAGAACCTACTGGATCATCATTGAATTAAAACACAAAGCAAGAGAAAAACCTTAT
GATGTTCAAAGTTTGCGGACTGCACTTGAGGAGGCGATCAAAACGCGTTATCAACTGGATCCAAA
ATTTATCACAAATATTTTGTATGAGGATAATGTTATCACTATTGATCTGGTTCAAAATTCTTCTC
AGAAAACTCAGAATGATGTGGACATAGCTGATGTGGCTTATTATTTTGAAAAAGATGTTAAAGGT
GAATCCTTGTTTCATTCTAAGAAAATGGACCTGAGAGTAAATGGGGAACAACTGGATCTGGATCC
TGGTCAAACTTTAATTTATTATGTCGATGAAAAAGCACCTGAATTCTCAATGCAGGGTCTAAAAG
CTGGTGTTATTGCTGTTATTGTGGTTGTGGTGATAGCAATTGTTGCTGGAATTGTTGTGCTGGTT
ATTTCCAGAAAGAAGAGAATGGCAAAGTATGAGAAGGCTGAGATAAAGGAGATGGGTGAGATGCA
TAGGGAACTCAATGCA 233. marmoset 1-27 artificial aa
QDGNEEMGDTTQNPYKVSISGTTVTLTDYKDDDDKTASFAAAQKECVCENYKLAVNCFLNDNGQC
CD3.epsilon.-EpCAM
QCTSIGAQNTVLCSKLAAKCLVMKAEMNGSKLGRRAKPEGALQNNDGLYDPDCDESGLFKAKQCN
GTSTCWCVNTAGVRRTDKDTEITCSERVRTYWIIIELKHKAREKPYDVQSLRTALEEAIKTRYQL
DPKFITNILYEDNVITIDLVQNSSQKTQNDVDIADVAYYFEKDVKGESLFHSKKMDLRVNGEQLD
LDPGQTLIYYVDEKAPEFSMQGLKAGVIAVIVVVVIAIVAGIVVLVISRKKRMAKYEKAEIKEMG
EMHRELNA 234. marmoset 1-27 artificial nt
ATGGGATGGAGCTGTATCATCCTCTTCTTGGTAGCAACAGCTACAGGTGTACACTCCCAGGACGG
CD3.epsilon.-EpCAM
TAATGAAGAAATGGGTGATACTACACAGAACCCATATAAAGTTTCCATCTCAGGAACCACAGTAA
CACTGACAGATTACAAGGACGACGATGACAAGACTGCGAGTTTTGCCGCAGCTCAGAAAGAATGT
GTCTGTGAAAACTACAAGCTGGCCGTAAACTGCTTTTTGAATGACAATGGTCAATGCCAGTGTAC
TTCGATTGGTGCACAAAATACTGTCCTTTGCTCAAAGCTGGCTGCCAAATGTTTGGTGATGAAGG
CAGAAATGAACGGCTCAAAACTTGGGAGAAGAGCGAAACCTGAAGGGGCTCTCCAGAACAATGAT
GGCCTTTACGATCCTGACTGCGATGAGAGCGGGCTCTTTAAGGCCAAGCAGTGCAACGGCACCTC
CACGTGCTGGTGTGTGAACACTGCTGGGGTCAGAAGAACTGACAAGGACACTGAAATAACCTGCT
CTGAGCGAGTGAGAACCTACTGGATCATCATTGAATTAAAACACAAAGCAAGAGAAAAACCTTAT
GATGTTCAAAGTTTGCGGACTGCACTTGAGGAGGCGATCAAAACGCGTTATCAACTGGATCCAAA
ATTTATCACAAATATTTTGTATGAGGATAATGTTATCACTATTGATCTGGTTCAAAATTCTTCTC
AGAAAACTCAGAATGATGTGGACATAGCTGATGTGGCTTATTATTTTGAAAAAGATGTTAAAGGT
GAATCCTTGTTTCATTCTAAGAAAATGGACCTGAGAGTAAATGGGGAACAACTGGATCTGGATCC
TGGTCAAACTTTAATTTATTATGTCGATGAAAAAGCACCTGAATTCTCAATGCAGGGTCTAAAAG
CTGGTGTTATTGCTGTTATTGTGGTTGTGGTGATAGCAATTGTTGCTGGAATTGTTGTGCTGGTT
ATTTCCAGAAAGAAGAGAATGGCAAAGTATGAGAAGGCTGAGATAAAGGAGATGGGTGAGATGCA
TAGGGAACTCAATGCA 235. tamarin 1-27 CD3.epsilon.- artificial aa
QDGNEEMGDTTQNPYKVSISGTTVTLTDYKDDDDKTASFAAAQKECVCENYKLAVNCFLNDNGQC
EpCAM
QCTSIGAQNTVLCSKLAAKCLVMKAEMNGSKLGRRAKPEGALQNNDGLYDPDCDESGLFKAKQCN
GTSTCWCVNTAGVRRTDKDTEITCSERVRTYWIIIELKHKAREKPYDVQSLRTALEEAIKTRYQL
DPKFITNILYEDNVITIDLVQNSSQKTQNDVDIADVAYYFEKDVKGESLFHSKKMDLRVNGEQLD
LDPGQTLIYYVDEKAPEFSMQGLKAGVIAVIVVVVIAIVAGIVVLVISRKKRMAKYEKAEIKEMG
EMHRELNA 236. tamarin 1-27 CD3.epsilon.- artificial nt
ATGGGATGGAGCTGTATCATCCTCTTCTTGGTAGCAACAGCTACAGGTGTACACTCCCAGGACGG
EpCAM
TAATGAAGAAATGGGTGATACTACACAGAACCCATATAAAGTTTCCATCTCAGGAACCACAGTAA
CACTGACAGATTACAAGGACGACGATGACAAGACTGCGAGTTTTGCCGCAGCTCAGAAAGAATGT
GTCTGTGAAAACTACAAGCTGGCCGTAAACTGCTTTTTGAATGACAATGGTCAATGCCAGTGTAC
TTCGATTGGTGCACAAAATACTGTCCTTTGCTCAAAGCTGGCTGCCAAATGTTTGGTGATGAAGG
CAGAAATGAACGGCTCAAAACTTGGGAGAAGAGCGAAACCTGAAGGGGCTCTCCAGAACAATGAT
GGCCTTTACGATCCTGACTGCGATGAGAGCGGGCTCTTTAAGGCCAAGCAGTGCAACGGCACCTC
CACGTGCTGGTGTGTGAACACTGCTGGGGTCAGAAGAACTGACAAGGACACTGAAATAACCTGCT
CTGAGCGAGTGAGAACCTACTGGATCATCATTGAATTAAAACACAAAGCAAGAGAAAAACCTTAT
GATGTTCAAAGTTTGCGGACTGCACTTGAGGAGGCGATCAAAACGCGTTATCAACTGGATCCAAA
ATTTATCACAAATATTTTGTATGAGGATAATGTTATCACTATTGATCTGGTTCAAAATTCTTCTC
AGAAAACTCAGAATGATGTGGACATAGCTGATGTGGCTTATTATTTTGAAAAAGATGTTAAAGGT
GAATCCTTGTTTCATTCTAAGAAAATGGACCTGAGAGTAAATGGGGAACAACTGGATCTGGATCC
TGGTCAAACTTTAATTTATTATGTCGATGAAAAAGCACCTGAATTCTCAATGCAGGGTCTAAAAG
CTGGTGTTATTGCTGTTATTGTGGTTGTGGTGATAGCAATTGTTGCTGGAATTGTTGTGCTGGTT
ATTTCCAGAAAGAAGAGAATGGCAAAGTATGAGAAGGCTGAGATAAAGGAGATGGGTGAGATGCA
TAGGGAACTCAATGCA 237. squirrel monkey 1-27 artificial aa
QDGNEEIGDTTQNPYKVSISGTTVTLTDYKDDDDKTASFAAAQKECVCENYKLAVNCFLNDNGQC
CD3.epsilon.-EpCAM
QCTSIGAQNTVLCSKLAAKCLVMKAEMNGSKLGRRAKPEGALQNNDGLYDPDCDESGLFKAKQCN
GTSTCWCVNTAGVRRTDKDTEITCSERVRTYWIIIELKHKAREKPYDVQSLRTALEEAIKTRYQL
DPKFITNILYEDNVITIDLVQNSSQKTQNDVDIADVAYYFEKDVKGESLFHSKKMDLRVNGEQLD
LDPGQTLIYYVDEKAPEFSMQGLKAGVIAVIVVVVIAIVAGIVVLVISRKKRMAKYEKAEIKEMG
EMHRELNA 238. squirrel monkey 1-27 artificial nt
ATGGGATGGAGCTGTATCATCCTCTTCTTGGTAGCAACAGCTACAGGTGTACACTCCCAGGACGG
CD3.epsilon.-EpCAM
TAATGAAGAGATTGGTGATACTACCCAGAACCCATATAAAGTTTCCATCTCAGGAACCACAGTAA
CACTGACAGATTACAAGGACGACGATGACAAGACTGCGAGTTTTGCCGCAGCTCAGAAAGAATGT
GTCTGTGAAAACTACAAGCTGGCCGTAAACTGCTTTTTGAATGACAATGGTCAATGCCAGTGTAC
TTCGATTGGTGCACAAAATACTGTCCTTTGCTCAAAGCTGGCTGCCAAATGTTTGGTGATGAAGG
CAGAAATGAACGGCTCAAAACTTGGGAGAAGAGCGAAACCTGAAGGGGCTCTCCAGAACAATGAT
GGCCTTTACGATCCTGACTGCGATGAGAGCGGGCTCTTTAAGGCCAAGCAGTGCAACGGCACCTC
CACGTGCTGGTGTGTGAACACTGCTGGGGTCAGAAGAACTGACAAGGACACTGAAATAACCTGCT
CTGAGCGAGTGAGAACCTACTGGATCATCATTGAATTAAAACACAAAGCAAGAGAAAAACCTTAT
GATGTTCAAAGTTTGCGGACTGCACTTGAGGAGGCGATCAAAACGCGTTATCAACTGGATCCAAA
ATTTATCACAAATATTTTGTATGAGGATAATGTTATCACTATTGATCTGGTTCAAAATTCTTCTC
AGAAAACTCAGAATGATGTGGACATAGCTGATGTGGCTTATTATTTTGAAAAAGATGTTAAAGGT
GAATCCTTGTTTCATTCTAAGAAAATGGACCTGAGAGTAAATGGGGAACAACTGGATCTGGATCC
TGGTCAAACTTTAATTTATTATGTCGATGAAAAAGCACCTGAATTCTCAATGCAGGGTCTAAAAG
CTGGTGTTATTGCTGTTATTGTGGTTGTGGTGATAGCAATTGTTGCTGGAATTGTTGTGCTGGTT
ATTTCCAGAAAGAAGAGAATGGCAAAGTATGAGAAGGCTGAGATAAAGGAGATGGGTGAGATGCA
TAGGGAACTCAATGCA 239. swine 1-27 CD3.epsilon.- artificial aa
QEDIERPDEDTQKTFKVSISGDKVELTDYKDDDDKTASFAAAQKECVCENYKLAVNCFLNDNGQC
EpCAM
QCTSIGAQNTVLCSKLAAKCLVMKAEMNGSKLGRRAKPEGALQNNDGLYDPDCDESGLFKAKQCN
GTSTCWCVNTAGVRRTDKDTEITCSERVRTYWIIIELKHKAREKPYDVQSLRTALEEAIKTRYQL
DPKFITNILYEDNVITIDLVQNSSQKTQNDVDIADVAYYFEKDVKGESLFHSKKMDLRVNGEQLD
LDPGQTLIYYVDEKAPEFSMQGLKAGVIAVIVVVVIAIVAGIVVLVISRKKRMAKYEKAEIKEMG
EMHRELNA 240. swine 1-27 CD3.epsilon.- artificial nt
ATGGGATGGAGCTGTATCATCCTCTTCTTGGTAGCAACAGCTACAGGTGTACACTCCCAAGAAGA
EpCAM
CATTGAAAGACCAGATGAAGATACACAGAAAACATTTAAAGTCTCCATCTCTGGAGACAAAGTAG
AGCTGACAGATTACAAGGACGACGATGACAAGACTGCGAGTTTTGCCGCAGCTCAGAAAGAATGT
GTCTGTGAAAACTACAAGCTGGCCGTAAACTGCTTTTTGAATGACAATGGTCAATGCCAGTGTAC
TTCGATTGGTGCACAAAATACTGTCCTTTGCTCAAAGCTGGCTGCCAAATGTTTGGTGATGAAGG
CAGAAATGAACGGCTCAAAACTTGGGAGAAGAGCGAAACCTGAAGGGGCTCTCCAGAACAATGAT
GGCCTTTACGATCCTGACTGCGATGAGAGCGGGCTCTTTAAGGCCAAGCAGTGCAACGGCACCTC
CACGTGCTGGTGTGTGAACACTGCTGGGGTCAGAAGAACTGACAAGGACACTGAAATAACCTGCT
CTGAGCGAGTGAGAACCTACTGGATCATCATTGAATTAAAACACAAAGCAAGAGAAAAACCTTAT
GATGTTCAAAGTTTGCGGACTGCACTTGAGGAGGCGATCAAAACGCGTTATCAACTGGATCCAAA
ATTTATCACAAATATTTTGTATGAGGATAATGTTATCACTATTGATCTGGTTCAAAATTCTTCTC
AGAAAACTCAGAATGATGTGGACATAGCTGATGTGGCTTATTATTTTGAAAAAGATGTTAAAGGT
GAATCCTTGTTTCATTCTAAGAAAATGGACCTGAGAGTAAATGGGGAACAACTGGATCTGGATCC
TGGTCAAACTTTAATTTATTATGTCGATGAAAAAGCACCTGAATTCTCAATGCAGGGTCTAAAAG
CTGGTGTTATTGCTGTTATTGTGGTTGTGGTGATAGCAATTGTTGCTGGAATTGTTGTGCTGGTT
ATTTCCAGAAAGAAGAGAATGGCAAAGTATGAGAAGGCTGAGATAAAGGAGATGGGTGAGATGCA
TAGGGAACTCAATGCA 241. human CD3 epsilon human aa
QDGNEEMGGITQTPYKVSISGTTVILTCPQYPGSEILWQHNDKNIGGDEDDKNIGSDEDHLSLKE
chain
FSELEQSGYYVCYPRGSKPEDANFYLYLRARVCENCMEMDVMSVATIVIVDICITGGLLLLVYYW
SKNRKAKAKPVTRGAGAGGRQRGQNKERPPPVPNPDYEPIRKGQRDLYSGLNQRRI 242. human
CD3 epsilon human nt
ATGCAGTCGGGCACTCACTGGAGAGTTCTGGGCCTCTGCCTCTTATCAGTTGGCGTTTGGGGGCA
chain
AGATGGTAATGAAGAAATGGGTGGTATTACACAGACACCATATAAAGTCTCCATCTCTGGAACCA
CAGTAATATTGACATGCCCTCAGTATCCTGGATCTGAAATACTATGGCAACACAATGATAAAAAC
ATAGGCGGTGATGAGGATGATAAAAACATAGGCAGTGATGAGGATCACCTGTCACTGAAGGAATT
TTCAGAATTGGAGCAAAGTGGTTATTATGTCTGCTACCCCAGAGGAAGCAAACCAGAAGATGCGA
ACTTTTATCTCTACCTGAGGGCACGCGTGTGTGAGAACTGCATGGAGATGGATGTGATGTCGGTG
GCCACAATTGTCATAGTGGACATCTGCATCACTGGGGGCTTGCTGCTGCTGGTTTACTACTGGAG
CAAGAATAGAAAGGCCAAGGCCAAGCCTGTGACACGAGGAGCGGGTGCTGGCGGCAGGCAAAGGG
GACAAAACAAGGAGAGGCCACCACCTGTTCCCAACCCAGACTATGAGCCCATCCGGAAAGGCCAG
CGGGACCTGTATTCTGGCCTGAATCAGAGACGCATC 243. 19 amino acid artificial
aa MGWSCIILFLVATATGVHS immunoglobulin leader peptide 244. 19 amino
acid artificial nt
ATGGGATGGAGCTGTATCATCCTCTTCTTGGTAGCAACAGCTACAGGTGTACACTCC
immunoglobulin leader peptide 245. murine IgG1 heavy murine aa
AKTTPPSVYPLAPGSAAQTNSMVTLGCLVKGYFPEPVTVTWNSGSLSSGVHTFPAVLQSDLYTLS
chain constant
SSVTVPSSTWPSETVTCNVAHPASSTKVDKKIVPRDCGCKPCICTVPEVSSVFIFPPKPKDVLTI
region
TLTPKVTCVVVDISKDDPEVQFSWFVDDVEVHTAQTQPREEQFNSTFRSVSELPIMHQDWLNGK- E
FKCRVNSAAFPAPIEKTISKTKGRPKAPQVYTIPPPKEQMAKDKVSLTCMITDFFPEDITVEWQW
NGQPAENYKNTQPIMDTDGSYFVYSKLNVQKSNWEAGNTFTCSVLHEGLHNHHTEKSLSHSPGK
246. murine IgG1 heavy murine nt
GCCAAAACGACACCCCCATCTGTCTATCCACTGGCCCCTGGATCTGCTGCCCAAACTAACTCCAT
chain constant
GGTGACCCTGGGATGCCTGGTCAAGGGCTATTTCCCTGAGCCAGTGACAGTGACCTGGAACTCTG
region
GATCCCTGTCCAGCGGTGTGCACACCTTCCCAGCTGTCCTGCAGTCTGACCTCTACACTCTGAG- C
AGCTCAGTGACTGTCCCCTCCAGCACCTGGCCCAGCGAGACCGTCACCTGCAACGTTGCCCACCC
GGCCAGCAGCACCAAGGTGGACAAGAAAATTGTGCCCAGGGATTGTGGTTGTAAGCCTTGCATAT
GTACAGTCCCAGAAGTATCATCTGTCTTCATCTTCCCCCCAAAGCCCAAGGATGTGCTCACCATT
ACTCTGACTCCTAAGGTCACGTGTGTTGTGGTAGACATCAGCAAGGATGATCCCGAGGTCCAGTT
CAGCTGGTTTGTAGATGATGTGGAGGTGCACACAGCTCAGACGCAACCCCGGGAGGAGCAGTTCA
ACAGCACTTTCCGCTCAGTCAGTGAACTTCCCATCATGCACCAGGACTGGCTCAATGGCAAGGAG
TTCAAATGCAGGGTCAACAGTGCAGCTTTCCCTGCCCCCATCGAGAAAACCATCTCCAAAACCAA
AGGCAGACCGAAGGCTCCACAGGTGTACACCATTCCACCTCCCAAGGAGCAGATGGCCAAGGATA
AAGTCAGTCTGACCTGCATGATAACAGACTTCTTCCCTGAAGACATTACTGTGGAGTGGCAGTGG
AATGGGCAGCCAGCGGAGAACTACAAGAACACTCAGCCCATCATGGACACAGATGGCTCTTACTT
CGTCTACAGCAAGCTCAATGTGCAGAAGAGCAACTGGGAGGCAGGAAATACTTTCACCTGCTCTG
TGTTACATGAGGGCCTGCACAACCACCATACTGAGAAGAGCCTCTCCCACTCTCCTGGTAAA 247.
human lambda light human aa
GQPKAAPSVTLFPPSSEELQANKATLVCLISDFYPGAVTVAWKADSSPVKAGVETTTPSKQSNNK
chain constant YAASSYLSLTPEQWKSHKSYSCQVTHEGSTVEKTVAPTECS region
248. human lambda light human nt
GGTCAGCCCAAGGCTGCCCCCTCGGTCACTCTGTTCCCACCCTCCTCTGAGGAGCTTCAAGCCAA
chain constant
CAAGGCCACACTGGTGTGTCTCATAAGTGACTTCTACCCGGGAGCCGTGACAGTGGCCTGGAAGG
region
CAGATAGCAGCCCCGTCAAGGCGGGAGTGGAGACCACCACACCCTCCAAACAAAGCAACAACAA- G
TACGCGGCCAGCAGCTACCTGAGCCTGACGCCTGAGCAGTGGAAGTCCCACAAAAGCTACAGCTG
CCAGGTCACGCATGAAGGGAGCACCGTGGAGAAGACAGTGGCCCCTACAGAATGTTCA 249.
c-terminal domain human aa
DYKDDDDKSRTRSGSQLDGGLVLFSHRGTLDGGFRFRLSDGEHTSPGHFFRVTAQKQVLLSLKGS
construct of human
QTLTVCPGSVQPLSSQTLRASSSAGTDPQLLLYRVVRGPQLGRLFHAQQDSTGEALVNFTQAEVY
MCSP
AGNILYEHEMPPEPFWEAHDTLELQLSSPPARDVAATLAVAVSFEAACPQRPSHLWKNKGLWVPE
GQRARITVAALDASNLLASVPSPQRSEHDVLFQVTQFPSRGQLLVSEEPLHAGQPHFLQSQLAAG
QLVYAHGGGGTQQDGFHFRAHLQGPAGASVAGPQTSEAFAITVRDVNERPPQPQASVPLRLTRGS
RAPISRAQLSVVDPDSAPGEIEYEVQRAPHNGFLSLVGGGLGPVTRFTQADVDSGRLAFVANGSS
VAGIFQLSMSDGASPPLPMSLAVDILPSAIEVQLRAPLEVPQALGRSSLSQQQLRVVSDREEPEA
AYRLIQGPQYGHLLVGGRPTSAFSQFQIDQGEVVFAFTNSSSSHDHFRVLALARGVNASAVVNVT
VRALLHVWAGGPWPQGATLRLDPTVLDAGELANRTDSVPRFRLLEGPRHGRVVRVPRARTEPGGS
QLVEQFTQQDLEDGRLGLEVGRPEGRAPGPAGDSLTLELWAQGVPPAVASLDFATEPYNAARPYS
VALLSVPEAARTEAGKPESSTPTGEPGPMASSPEPAVAKGGFLSFLEANMFSVIIPMCLVLLLLA
LILPLLFYLRKRNKTGKHDVQVLTAKPRNGLAGDTETFRKVEPGQAIPLTAVPGQGPPPGGQPDP
ELLQFCRTPNPALKNGQYWV 250. c-terminal domain human nt
ATGGGATGGAGCTGTATCATCCTCTTCTTGGTAGCAACAGCTACAGGTGTACACTCCGACTACAA
construct of human
AGACGATGACGACAAGTCCCGTACGAGATCTGGATCCCAATTGGACGGCGGGCTCGTGCTGTTCT
MCSP
CACACAGAGGAACCCTGGATGGAGGCTTCCGCTTCCGCCTCTCTGACGGCGAGCACACTTCCCCC
GGACACTTCTTCCGAGTGACGGCCCAGAAGCAAGTGCTCCTCTCGCTGAAGGGCAGCCAGACACT
GACTGTCTGCCCAGGGTCCGTCCAGCCACTCAGCAGTCAGACCCTCAGGGCCAGCTCCAGCGCAG
GCACTGACCCCCAGCTCCTGCTCTACCGTGTGGTGCGGGGCCCCCAGCTAGGCCGGCTGTTCCAC
GCCCAGCAGGACAGCACAGGGGAGGCCCTGGTGAACTTCACTCAGGCAGAGGTCTACGCTGGGAA
TATTCTGTATGAGCATGAGATGCCCCCCGAGCCCTTTTGGGAGGCCCATGATACCCTAGAGCTCC
AGCTGTCCTCGCCGCCTGCCCGGGACGTGGCCGCCACCCTTGCTGTGGCTGTGTCTTTTGAGGCT
GCCTGTCCCCAGCGCCCCAGCCACCTCTGGAAGAACAAAGGTCTCTGGGTCCCCGAGGGCCAGCG
GGCCAGGATCACCGTGGCTGCTCTGGATGCCTCCAATCTCTTGGCCAGCGTTCCATCACCCCAGC
GCTCAGAGCATGATGTGCTCTTCCAGGTCACACAGTTCCCCAGCCGCGGCCAGCTGTTGGTGTCC
GAGGAGCCCCTCCATGCTGGGCAGCCCCACTTCCTGCAGTCCCAGCTGGCTGCAGGGCAGCTAGT
GTATGCCCACGGCGGTGGGGGCACCCAGCAGGATGGCTTCCACTTTCGTGCCCACCTCCAGGGGC
CAGCAGGGGCCTCCGTGGCTGGACCCCAAACCTCAGAGGCCTTTGCCATCACGGTGAGGGATGTA
AATGAGCGGCCCCCTCAGCCACAGGCCTCTGTCCCACTCCGGCTCACCCGAGGCTCTCGTGCCCC
CATCTCCCGGGCCCAGCTGAGTGTGGTGGACCCAGACTCAGCTCCTGGGGAGATTGAGTACGAGG
TCCAGCGGGCACCCCACAACGGCTTCCTCAGCCTGGTGGGTGGTGGCCTGGGGCCCGTGACCCGC
TTCACGCAAGCCGATGTGGATTCAGGGCGGCTGGCCTTCGTGGCCAACGGGAGCAGCGTGGCAGG
CATCTTCCAGCTGAGCATGTCTGATGGGGCCAGCCCACCCCTGCCCATGTCCCTGGCTGTGGACA
TCCTACCATCCGCCATCGAGGTGCAGCTGCGGGCACCCCTGGAGGTGCCCCAAGCTTTGGGGCGC
TCCTCACTGAGCCAGCAGCAGCTCCGGGTGGTTTCAGATCGGGAGGAGCCAGAGGCAGCATACCG
GTTGATCCAGGGACCCCAGTATGGGCATCTCCTGGTGGGCGGGCGGCCCACCTCGGCCTTCAGCC
AATTCCAGATAGACCAGGGCGAGGTGGTCTTTGCCTTCACCAACTCCTCCTCCTCTCATGACCAC
TTCAGAGTCCTGGCACTGGCTAGGGGTGTCAATGCATCAGCCGTAGTGAACGTCACTGTGAGGGC
TCTGCTGCATGTGTGGGCAGGTGGGCCATGGCCCCAGGGTGCCACCCTGCGCCTGGACCCCACCG
TCCTAGATGCTGGCGAGCTGGCCAACCGCACAGACAGTGTGCCGCGCTTCCGCCTCCTGGAGGGA
CCCCGGCATGGCCGCGTGGTCCGCGTGCCCCGAGCCAGGACGGAGCCCGGGGGCAGCCAGCTGGT
GGAGCAGTTCACTCAGCAGGACCTTGAGGACGGGAGGCTGGGGCTGGAGGTGGGCAGGCCAGAGG
GGAGGGCCCCCGGCCCCGCAGGTGACAGTCTCACTCTGGAGCTGTGGGCACAGGGCGTCCCGCCT
GCTGTGGCCTCCCTGGACTTTGCCACTGAGCCTTACAATGCTGCCCGGCCCTACAGCGTGGCCCT
GCTCAGTGTCCCCGAGGCCGCCCGGACGGAAGCAGGGAAGCCAGAGAGCAGCACCCCCACAGGCG
AGCCAGGCCCCATGGCATCCAGCCCTGAGCCCGCTGTGGCCAAGGGAGGCTTCCTGAGCTTTCTA
GAGGCCAACATGTTCAGCGTCATCATCCCCATGTGCCTGGTACTTCTGCTCCTGGCGCTCATCCT
GCCCCTGCTCTTCTACCTCCGAAAACGCAACAAGACGGGCAAGCATGACGTCCAGGTCCTGACTG
CCAAGCCCCGCAACGGCCTGGCTGGTGACACCGAGACCTTTCGCAAGGTGGAGCCAGGCCAGGCC
ATCCCGCTCACAGCTGTGCCTGGCCAGGGGCCCCCTCCAGGAGGCCAGCCTGACCCAGAGCTGCT
GCAGTTCTGCCGGACACCCAACCCTGCCCTTAAGAATGGCCAGTACTGGGTG 251. partial
sequence of cynomolgus aa
PSNGRVVLRAAPGTEVRSFTQAQLDGGLVLFSHRGTLDGGFRFGLSDGEHTSSGHFFRVTAQKQV
cynomolgus MCSP
LLSLEGSRTLTVCPGSVQPLSSQTLRASSSAGTDPQLLLYRVVRGPQLGRLFHAQQDSTGEALVN
FTQAEVYAGNILYEHEMPTEPFWEAHDTLELQLSSPPARDVAATLAVAVSFEAACPQRPSHLWKN
KGLWVPEGQRAKITMAALDASNLLASVPSSQRLEHDVLFQVTQFPSRGQLLVSEEPLHAGQPHFL
QSQLAAGQLVYAHGGGGTQQDGFHFRAHLQGPAGATVAGPQTSEAFAITVRDVNERPPQPQASVP
LRITRGSRAPISRAQLSVVDPDSAPGEIEYEVQRAPHNGFLSLVGGGPGPVNRFTQADVDSGRLA
FVANGSSVAGVFQLSMSDGASPPLPMSLAVDILPSAIEVQLQAPLEVPQALGRSSLSQQQLRVVS
DREEPEAAYRLIQGPKYGHLLVGGQPASAFSQLQIDQGEVVFAFTNFSSSHDHFRVLALARGVNA
SAVVNITVRALLHVWAGGPWPQGATLRLDPTILDAGELANRTGSVPRFRLLEGPRHGRVVRVPRA
RMEPGGSQLVEQFTQQDLEDGRLGLEVGRPEGRAPSPTGDSLTLELWAQGVPPAVASLDFATEPY
NAARPYSVALLSVPEATRTEAGKPESSTPTGEPGPMASSPVPAVAKGGFLGFLEANMFSVIIPXC
LVLLLLALILPLLFYLRKRNKTGKHDVQVLTAKPRNGLAGDTETFRKVEPGQAIPLTAVPGQGPP
PGGQPDPELLQFCRTPNPALKNGQYWV 252. partial sequence of cynomolgus nt
CCCAGCAACGGACGGGTAGTGCTGCGGGCGGCGCCGGGCACCGAGGTGCGCAGCTTCACGCAGGC
cynomolgus MCSP
CCAGCTGGATGGCGGACTCGTGCTGTTCTCACACAGAGGAACCCTGGATGGAGGCTTCCGCTTCG
GCCTCTCCGATGGCGAGCACACTTCCTCTGGACACTTCTTCCGAGTGACGGCCCAGAAGCAAGTG
CTCCTCTCGCTGGAGGGCAGCCGGACACTGACTGTCTGCCCAGGGTCCGTGCAGCCACTCAGCAG
TCAGACCCTCAGAGCCAGCTCCAGCGCAGGCACCGACCCCCAGCTCCTGCTCTACCGTGTGGTGC
GGGGCCCCCAGCTAGGCCGGCTGTTCCATGCCCAGCAGGACAGCACAGGGGAGGCCCTGGTGAAC
TTCACTCAGGCAGAGGTCTATGCTGGGAATATTCTGTATGAGCATGAGATGCCCACCGAGCCCTT
CTGGGAGGCCCATGATACCCTAGAGCTCCAGCTGTCCTCACCACCTGCCCGGGACGTGGCTGCCA
CCCTTGCTGTGGCTGTGTCTTTTGAGGCTGCCTGTCCCCAGCGCCCCAGCCACCTCTGGAAGAAC
AAAGGTCTCTGGGTCCCCGAGGGCCAGCGGGCCAAGATCACCATGGCTGCCCTGGATGCCTCCAA
CCTCTTGGCCAGCGTTCCATCATCCCAGCGCCTAGAGCATGATGTGCTCTTCCAGGTCACGCAGT
TCCCCAGCCGGGGCCAGCTATTGGTGTCTGAGGAGCCCCTCCACGCTGGGCAGCCCCACTTCCTG
CAGTCCCAGCTGGCTGCAGGGCAGCTAGTGTATGCCCACGGCGGTGGGGGTACCCAACAGGATGG
CTTCCACTTTCGTGCCCACCTCCAGGGGCCAGCAGGGGCCACCGTGGCTGGACCCCAAACCTCAG
AGGCTTTTGCCATCACGGTGCGGGATGTAAATGAGCGGCCCCCTCAGCCACAGGCCTCTGTCCCA
CTCCGGATCACCCGAGGCTCTCGAGCCCCCATCTCCCGGGCCCAGCTGAGTGTCGTGGACCCAGA
CTCAGCTCCTGGGGAGATTGAGTATGAGGTCCAGCGGGCACCCCACAACGGCTTCCTCAGCCTGG
TGGGTGGTGGCCCGGGGCCCGTGAACCGCTTCACGCAAGCCGATGTGGATTCGGGGCGGCTGGCC
TTCGTGGCCAACGGGAGCAGCGTAGCAGGCGTCTTCCAGCTGAGCATGTCTGATGGGGCCAGCCC
ACCGCTGCCCATGTCCCTGGCCGTGGACATCCTACCATCCGCCATCGAGGTGCAGCTGCAGGCAC
CCCTGGAGGTGCCCCAAGCTTTGGGGCGCTCCTCACTGAGCCAGCAGCAGCTCCGGGTGGTTTCA
GATAGGGAGGAGCCAGAGGCAGCATACCGCCTCATCCAGGGACCAAAGTACGGGCATCTCCTGGT
GGGTGGGCAGCCCGCCTCGGCCTTCAGCCAACTCCAGATAGACCAGGGCGAGGTGGTCTTTGCCT
TCACCAACTTCTCCTCCTCTCATGACCACTTCAGAGTCCTGGCACTGGCTAGGGGTGTCAACGCA
TCAGCCGTAGTGAACATCACTGTGAGGGCTCTGCTGCACGTGTGGGCAGGTGGGCCATGGCCCCA
GGGTGCTACCCTGCGCCTGGACCCAACCATCCTAGATGCTGGCGAGCTGGCCAACCGCACAGGCA
GTGTGCCCCGCTTCCGCCTCCTGGAGGGACCCCGGCATGGCCGCGTGGTCCGTGTGCCCCGAGCC
AGGATGGAGCCTGGGGGCAGCCAGCTGGTGGAGCAGTTCACTCAGCAGGACCTTGAGGATGGGAG
GCTGGGGCTGGAGGTGGGCAGGCCAGAGGGAAGGGCCCCCAGCCCCACAGGCGACAGTCTCACTC
TGGAGCTGTGGGCACAGGGCGTCCCACCTGCTGTGGCCTCCCTGGACTTTGCCACTGAGCCTTAC
AATGCTGCCCGGCCCTACAGCGTGGCCCTGCTCAGTGTCCCCGAGGCCACCCGGACGGAAGCAGG
GAAGCCAGAGAGCAGCACCCCCACAGGCGAGCCAGGCCCCATGGCATCTAGCCCTGTGCCTGCTG
TGGCCAAGGGAGGCTTCCTGGGCTTCCTTGAGGCCAACATGTTCAGTGTCATCATCCCCRTGTGC
CTGGTCCTTCTGCTCCTGGCGCTCATCTTGCCCCTGCTCTTCTACCTCCGAAAACGCAACAAGAC
GGGCAAGCATGACGTCCAGGTCCTGACTGCCAAGCCCCGCAATGGTCTGGCTGGTGACACTGAGA
CCTTTCGCAAGGTGGAGCCAGGCCAGGCCATCCCGCTCACAGCTGTGCCTGGCCAGGGGCCCCCT
CCGGGAGGCCAGCCTGACCCAGAGCTGCTGCAGTTCTGCCGGACACCCAACCCTGCCCTTAAGAA
TGGCCAGTACTGGGTG 253. PCR primer for CD3.epsilon. artificial nt
AGAGTTCTGGGCCTCTGC chain - forward primer 254. PCR primer for
CD3.epsilon. artificial nt CGGATGGGCTCATAGTCTG chain - reverse
primer 255. His6-human CD3.epsilon. artificial aa
HHHHHHQDGNEEMGGITQTPYKVSISGTTVILTCPQYPGSEILWQHNDKNIGGDEDDKNIGSDED
HLSLKEFSELEQSGYYVCYPRGSKPEDANFYLYLRARVCENCMEMDVMSVATIVIVDICITGGLL
LLVYYWSKNRKAKAKPVTRGAGAGGRQRGQNKERPPPVPNPDYEPIRKGQRDLYSGLNQRRI 256.
His6-human CD3.epsilon. artificial nt
ATGGGATGGAGCTGTATCATCCTCTTCTTGGTAGCAACAGCTACAGGTGTACACTCCCATCATCA
CCATCATCATCAAGATGGTAATGAAGAAATGGGTGGTATTACACAGACACCATATAAAGTCTCCA
TCTCTGGAACCACAGTAATATTGACATGCCCTCAGTATCCTGGATCTGAAATACTATGGCAACAC
AATGATAAAAACATAGGCGGTGATGAGGATGATAAAAACATAGGCAGTGATGAGGATCACCTGTC
ACTGAAGGAATTTTCAGAATTGGAGCAAAGTGGTTATTATGTCTGCTACCCCAGAGGAAGCAAAC
CAGAAGATGCGAACTTTTATCTCTACCTGAGGGCACGCGTGTGTGAGAACTGCATGGAGATGGAT
GTGATGTCGGTGGCCACAATTGTCATAGTGGACATCTGCATCACTGGGGGCTTGCTGCTGCTGGT
TTACTACTGGAGCAAGAATAGAAAGGCCAAGGCCAAGCCTGTGACACGAGGAGCGGGTGCTGGCG
GCAGGCAAAGGGGACAAAACAAGGAGAGGCCACCACCTGTTCCCAACCCAGACTATGAGCCCATC
CGGAAAGGCCAGCGGGACCTGTATTCTGGCCTGAATCAGAGACGCATC 257. CD33 AH3 HL
.times. H2C artificial aa
QVQLVQSGAEVKKPGESVKVSCKASGYTFTNYGMNWVRQAPGQGLEWMGWINTYTGEPTYADDFK
HL
GRVTMSSDTSTSTAYLEINSLRSDDTAIYYCARWSWSDGYYVYFDYWGQGTTVTVSSggggsggg
gsggggsDIVMTQSPDSLTVSLGERTTINCKSSQSVLDSSKNKNSLAWYQQKPGQPPKLLLSWAS
TRESGIPDRFSGSGSGTDFTLTIDSLQPEDSATYYCQQSAHFPITFGQGTRLEIKSGGGGSEVQL
VESGGGLVQPGGSLKLSCAASGFTFNKYAMNWVRQAPGKGLEWVARIRSKYNNYATYYADSVKDR
FTISRDDSKNTAYLQMNNLKTEDTAVYYCVRHGNFGNSYISYWAYWGQGTLVTVSSGGGGSGGGG
SGGGGSQTVVTQEPSLTVSPGGTVTLTCGSSTGAVTSGYYPNWVQQKPGQAPRGLIGGTKFLAPG
TPARFSGSLLGGKAALTLSGVQPEDEAEYYCALWYSNRWVFGGGTKLTVL 258. CD33 AH3 HL
.times. H2C artificial nt
CAGGTGCAGCTGGTGCAGTCTGGAGCTGAGGTGAAAAAGCCTGGAGAGTCAGTCAAGGTCTCCTG
HL
CAAGGCTAGCGGGTATACCTTCACAAACTATGGAATGAACTGGGTGAGGCAGGCTCCAGGACAGG
GTTTAGAGTGGATGGGCTGGATAAACACCTACACTGGAGAGCCAACATATGCTGATGACTTCAAG
GGACGGGTTACCATGTCTTCGGATACCTCTACCAGCACTGCCTATTTGGAAATCAACAGCCTCAG
AAGTGATGACACGGCTATATATTACTGTGCGCGCTGGAGTTGGAGTGATGGTTACTACGTTTACT
TTGACTACTGGGGCCAAGGCACTACGGTCACCGTCTCCTCAGGTGGTGGTGGTTCTGGCGGCGGC
GGCTCCGGTGGTGGTGGTTCTGACATCGTGATGACACAGTCTCCAGACTCCCTGACTGTGTCTCT
GGGCGAGAGGACCACCATCAACTGCAAGTCCAGCCAGAGTGTTTTAGACAGCTCCAAGAATAAGA
ACTCCTTAGCTTGGTACCAGCAGAAACCAGGACAGCCTCCTAAATTACTCCTTTCCTGGGCATCT
ACGCGGGAATCCGGGATCCCTGACCGATTCAGTGGCAGCGGGTCTGGGACAGATTTCACTCTCAC
TATTGACAGCCTGCAGCCTGAAGATTCTGCAACTTACTATTGTCAACAGTCTGCCCACTTCCCGA
TCACCTTTGGCCAAGGGACACGACTGGAGATTAAATCCGGAGGTGGTGGCTCCGAGGTGCAGCTG
GTCGAGTCTGGAGGAGGATTGGTGCAGCCTGGAGGGTCATTGAAACTCTCATGTGCAGCCTCTGG
ATTCACCTTCAATAAGTACGCCATGAACTGGGTCCGCCAGGCTCCAGGAAAGGGTTTGGAATGGG
TTGCTCGCATAAGAAGTAAATATAATAATTATGCAACATATTATGCCGATTCAGTGAAAGACAGG
TTCACCATCTCCAGAGATGATTCAAAAAACACTGCCTATCTACAAATGAACAACTTGAAAACTGA
GGACACTGCCGTGTACTACTGTGTGAGACATGGGAACTTCGGTAATAGCTACATATCCTACTGGG
CTTACTGGGGCCAAGGGACTCTGGTCACCGTCTCCTCAGGTGGTGGTGGTTCTGGCGGCGGCGGC
TCCGGTGGTGGTGGTTCTCAGACTGTTGTGACTCAGGAACCTTCACTCACCGTATCACCTGGTGG
AACAGTCACACTCACTTGTGGCTCCTCGACTGGGGCTGTTACATCTGGCTACTACCCAAACTGGG
TCCAACAAAAACCAGGTCAGGCACCCCGTGGTCTAATAGGTGGGACTAAGTTCCTCGCCCCCGGT
ACTCCTGCCAGATTCTCAGGCTCCCTGCTTGGAGGCAAGGCTGCCCTCACCCTCTCAGGGGTACA
GCCAGAGGATGAGGCAGAATATTACTGTGCTCTATGGTACAGCAACCGCTGGGTGTTCGGTGGAG
GAACCAAACTGACTGTCCTA 259. CD33 AH3 HL .times. artificial aa
QVQLVQSGAEVKKPGESVKVSCKASGYTFTNYGMNWVRQAPGQGLEWMGWINTYTGEPTYADDFK
F12Q HL
GRVTMSSDTSTSTAYLEINSLRSDDTAIYYCARWSWSDGYYVYFDYWGQGTTVTVSSggggsg- gg
gsggggsDIVMTQSPDSLTVSLGERTTINCKSSQSVLDSSKNKNSLAWYQQKPGQPPKLLLSWAS
TRESGIPDRFSGSGSGTDFTLTIDSLQPEDSATYYCQQSAHFPITFGQGTRLEIKSGGGGSEVQL
VESGGGLVQPGGSLKLSCAASGFTFNSYAMNWVRQAPGKGLEWVARIRSKYNNYATYYADSVKGR
FTISRDDSKNTAYLQMNNLKTEDTAVYYCVRHGNFGNSYVSWWAYWGQGTLVTVSSGGGGSGGGG
SGGGGSQTVVTQEPSLTVSPGGTVTLTCGSSTGAVTSGNYPNWVQQKPGQAPRGLIGGTKFLAPG
TPARFSGSLLGGKAALTLSGVQPEDEAEYYCVLWYSNRWVFGGGTKLTVL 260. CD33 AH3 HL
.times. artificial nt
CAGGTGCAGCTGGTGCAGTCTGGAGCTGAGGTGAAAAAGCCTGGAGAGTCAGTCAAGGTCTCCTG
F12Q HL
CAAGGCTAGCGGGTATACCTTCACAAACTATGGAATGAACTGGGTGAGGCAGGCTCCAGGACA- GG
GTTTAGAGTGGATGGGCTGGATAAACACCTACACTGGAGAGCCAACATATGCTGATGACTTCAAG
GGACGGGTTACCATGTCTTCGGATACCTCTACCAGCACTGCCTATTTGGAAATCAACAGCCTCAG
AAGTGATGACACGGCTATATATTACTGTGCGCGCTGGAGTTGGAGTGATGGTTACTACGTTTACT
TTGACTACTGGGGCCAAGGCACTACGGTCACCGTCTCCTCAGGTGGTGGTGGTTCTGGCGGCGGC
GGCTCCGGTGGTGGTGGTTCTGACATCGTGATGACACAGTCTCCAGACTCCCTGACTGTGTCTCT
GGGCGAGAGGACCACCATCAACTGCAAGTCCAGCCAGAGTGTTTTAGACAGCTCCAAGAATAAGA
ACTCCTTAGCTTGGTACCAGCAGAAACCAGGACAGCCTCCTAAATTACTCCTTTCCTGGGCATCT
ACGCGGGAATCCGGGATCCCTGACCGATTCAGTGGCAGCGGGTCTGGGACAGATTTCACTCTCAC
TATTGACAGCCTGCAGCCTGAAGATTCTGCAACTTACTATTGTCAACAGTCTGCCCACTTCCCGA
TCACCTTTGGCCAAGGGACACGACTGGAGATTAAATCCGGAGGTGGTGGCTCCGAGGTGCAGCTG
GTCGAGTCTGGAGGAGGATTGGTGCAGCCTGGAGGGTCATTGAAACTCTCATGTGCAGCCTCTGG
ATTCACCTTCAATAGCTACGCCATGAACTGGGTCCGCCAGGCTCCAGGAAAGGGTTTGGAATGGG
TTGCTCGCATAAGAAGTAAATATAATAATTATGCAACATATTATGCCGATTCAGTGAAAGGCAGG
TTCACCATCTCCAGAGATGATTCAAAAAACACTGCCTATCTACAAATGAACAACTTGAAAACTGA
GGACACTGCCGTGTACTACTGTGTGAGACATGGGAACTTCGGTAATAGCTACGTTTCCTGGTGGG
CTTACTGGGGCCAAGGGACTCTGGTCACCGTCTCCTCAGGTGGTGGTGGTTCTGGCGGCGGCGGC
TCCGGTGGTGGTGGTTCTCAGACTGTTGTGACTCAGGAACCTTCACTCACCGTATCACCTGGTGG
AACAGTCACACTCACTTGTGGCTCCTCGACTGGGGCTGTTACATCTGGCAACTACCCAAACTGGG
TCCAACAAAAACCAGGTCAGGCACCCCGTGGTCTAATAGGTGGGACTAAGTTCCTCGCCCCCGGT
ACTCCTGCCAGATTCTCAGGCTCCCTGCTTGGAGGCAAGGCTGCCCTCACCCTCTCAGGGGTACA
GCCAGAGGATGAGGCAGAATATTACTGTGTTCTATGGTACAGCAACCGCTGGGTGTTCGGTGGAG
GAACCAAACTGACTGTCCTA 261. CD33 AH3 HL .times. I2C artificial aa
QVQLVQSGAEVKKPGESVKVSCKASGYTFTNYGMNWVRQAPGQGLEWMGWINTYTGEPTYADDFK
HL
GRVTMSSDTSTSTAYLEINSLRSDDTAIYYCARWSWSDGYYVYFDYWGQGTTVTVSSggggsggg
gsggggsDIVMTQSPDSLTVSLGERTTINCKSSQSVLDSSKNKNSLAWYQQKPGQPPKLLLSWAS
TRESGIPDRFSGSGSGTDFTLTIDSLQPEDSATYYCQQSAHFPITFGQGTRLEIKSGGGGSEVQL
VESGGGLVQPGGSLKLSCAASGFTFNKYAMNWVRQAPGKGLEWVARIRSKYNNYATYYADSVKDR
FTISRDDSKNTAYLQMNNLKTEDTAVYYCVRHGNFGNSYISYWAYWGQGTLVTVSSGGGGSGGGG
SGGGGSQTVVTQEPSLTVSPGGTVTLTCGSSTGAVTSGNYPNWVQQKPGQAPRGLIGGTKFLAPG
TPARFSGSLLGGKAALTLSGVQPEDEAEYYCVLWYSNRWVFGGGTKLTVL 262. CD33 AH3 HL
.times. I2C artificial nt
CAGGTGCAGCTGGTGCAGTCTGGAGCTGAGGTGAAAAAGCCTGGAGAGTCAGTCAAGGTCTCCTG
HL
CAAGGCTAGCGGGTATACCTTCACAAACTATGGAATGAACTGGGTGAGGCAGGCTCCAGGACAGG
GTTTAGAGTGGATGGGCTGGATAAACACCTACACTGGAGAGCCAACATATGCTGATGACTTCAAG
GGACGGGTTACCATGTCTTCGGATACCTCTACCAGCACTGCCTATTTGGAAATCAACAGCCTCAG
AAGTGATGACACGGCTATATATTACTGTGCGCGCTGGAGTTGGAGTGATGGTTACTACGTTTACT
TTGACTACTGGGGCCAAGGCACTACGGTCACCGTCTCCTCAGGTGGTGGTGGTTCTGGCGGCGGC
GGCTCCGGTGGTGGTGGTTCTGACATCGTGATGACACAGTCTCCAGACTCCCTGACTGTGTCTCT
GGGCGAGAGGACCACCATCAACTGCAAGTCCAGCCAGAGTGTTTTAGACAGCTCCAAGAATAAGA
ACTCCTTAGCTTGGTACCAGCAGAAACCAGGACAGCCTCCTAAATTACTCCTTTCCTGGGCATCT
ACGCGGGAATCCGGGATCCCTGACCGATTCAGTGGCAGCGGGTCTGGGACAGATTTCACTCTCAC
TATTGACAGCCTGCAGCCTGAAGATTCTGCAACTTACTATTGTCAACAGTCTGCCCACTTCCCGA
TCACCTTTGGCCAAGGGACACGACTGGAGATTAAATCCGGAGGTGGTGGCTCCGAGGTGCAGCTG
GTCGAGTCTGGAGGAGGATTGGTGCAGCCTGGAGGGTCATTGAAACTCTCATGTGCAGCCTCTGG
ATTCACCTTCAATAAGTACGCCATGAACTGGGTCCGCCAGGCTCCAGGAAAGGGTTTGGAATGGG
TTGCTCGCATAAGAAGTAAATATAATAATTATGCAACATATTATGCCGATTCAGTGAAAGACAGG
TTCACCATCTCCAGAGATGATTCAAAAAACACTGCCTATCTACAAATGAACAACTTGAAAACTGA
GGACACTGCCGTGTACTACTGTGTGAGACATGGGAACTTCGGTAATAGCTACATATCCTACTGGG
CTTACTGGGGCCAAGGGACTCTGGTCACCGTCTCCTCAGGTGGTGGTGGTTCTGGCGGCGGCGGC
TCCGGTGGTGGTGGTTCTCAGACTGTTGTGACTCAGGAACCTTCACTCACCGTATCACCTGGTGG
AACAGTCACACTCACTTGTGGCTCCTCGACTGGGGCTGTTACATCTGGCAACTACCCAAACTGGG
TCCAACAAAAACCAGGTCAGGCACCCCGTGGTCTAATAGGTGGGACTAAGTTCCTCGCCCCCGGT
ACTCCTGCCAGATTCTCAGGCTCCCTGCTTGGAGGCAAGGCTGCCCTCACCCTCTCAGGGGTACA
GCCAGAGGATGAGGCAGAATATTACTGTGTTCTATGGTACAGCAACCGCTGGGTGTTCGGTGGAG
GAACCAAACTGACTGTCCTA 263. CD33 AF5 HL .times. H2C artificial aa
QVQLVQSGAEVKKPGASVKVSCKASGYTFTNYGMNWVKQAPGQGLKWMGWINTYTGEPTYADDFK
HL
GRVTMTSDTSTSTAYLELHNLRSDDTAVYYCARWSWSDGYYVYFDYWGQGTTVTVSSggggsggg
gsggggsDIVMTQSPDSLTVSLGERTTINCKSSQSVLDSSKNKNSLAWYQQKPGQPPKLLLSWAS
TRESGIPDRFSGSGSGTDFTLTIDSLQPEDSATYYCQQSAHFPITFGQGTRLEIKSGGGGSEVQL
VESGGGLVQPGGSLKLSCAASGFTFNKYAMNWVRQAPGKGLEWVARIRSKYNNYATYYADSVKDR
FTISRDDSKNTAYLQMNNLKTEDTAVYYCVRHGNFGNSYISYWAYWGQGTLVTVSSGGGGSGGGG
SGGGGSQTVVTQEPSLTVSPGGTVTLTCGSSTGAVTSGYYPNWVQQKPGQAPRGLIGGTKFLAPG
TPARFSGSLLGGKAALTLSGVQPEDEAEYYCALWYSNRWVFGGGTKLTVL 264. CD33 AF5 HL
.times. H2C artificial nt
CAGGTGCAGCTGGTGCAGTCTGGAGCTGAGGTGAAGAAGCCTGGAGCGTCAGTCAAGGTCTCCTG
HL
CAAGGCTAGCGGGTATACCTTCACAAACTATGGAATGAACTGGGTGAAGCAGGCTCCAGGACAGG
GTTTAAAGTGGATGGGCTGGATAAACACCTACACTGGAGAGCCAACATATGCTGATGACTTCAAG
GGACGGGTTACCATGACTTCGGATACCTCTACCAGCACTGCCTATTTGGAACTCCACAACCTCAG
AAGTGATGACACGGCTGTATATTACTGTGCGCGCTGGAGTTGGAGTGATGGTTACTACGTTTACT
TTGACTACTGGGGCCAAGGCACTACGGTCACCGTCTCCTCAGGTGGTGGTGGTTCTGGCGGCGGC
GGCTCCGGTGGTGGTGGTTCTGACATCGTGATGACACAGTCTCCAGACTCCCTGACTGTGTCTCT
GGGCGAGAGGACCACCATCAACTGCAAGTCCAGCCAGAGTGTTTTAGACAGCTCCAAGAATAAGA
ACTCCTTAGCTTGGTACCAGCAGAAACCAGGACAGCCTCCTAAATTACTCCTTTCCTGGGCATCT
ACGCGGGAATCCGGGATCCCTGACCGATTCAGTGGCAGCGGGTCTGGGACAGATTTCACTCTCAC
TATTGACAGCCTGCAGCCTGAAGATTCTGCAACTTACTATTGTCAACAGTCTGCCCACTTCCCGA
TCACCTTTGGCCAAGGGACACGACTGGAGATTAAATCCGGAGGTGGTGGCTCCGAGGTGCAGCTG
GTCGAGTCTGGAGGAGGATTGGTGCAGCCTGGAGGGTCATTGAAACTCTCATGTGCAGCCTCTGG
ATTCACCTTCAATAAGTACGCCATGAACTGGGTCCGCCAGGCTCCAGGAAAGGGTTTGGAATGGG
TTGCTCGCATAAGAAGTAAATATAATAATTATGCAACATATTATGCCGATTCAGTGAAAGACAGG
TTCACCATCTCCAGAGATGATTCAAAAAACACTGCCTATCTACAAATGAACAACTTGAAAACTGA
GGACACTGCCGTGTACTACTGTGTGAGACATGGGAACTTCGGTAATAGCTACATATCCTACTGGG
CTTACTGGGGCCAAGGGACTCTGGTCACCGTCTCCTCAGGTGGTGGTGGTTCTGGCGGCGGCGGC
TCCGGTGGTGGTGGTTCTCAGACTGTTGTGACTCAGGAACCTTCACTCACCGTATCACCTGGTGG
AACAGTCACACTCACTTGTGGCTCCTCGACTGGGGCTGTTACATCTGGCTACTACCCAAACTGGG
TCCAACAAAAACCAGGTCAGGCACCCCGTGGTCTAATAGGTGGGACTAAGTTCCTCGCCCCCGGT
ACTCCTGCCAGATTCTCAGGCTCCCTGCTTGGAGGCAAGGCTGCCCTCACCCTCTCAGGGGTACA
GCCAGAGGATGAGGCAGAATATTACTGTGCTCTATGGTACAGCAACCGCTGGGTGTTCGGTGGAG
GAACCAAACTGACTGTCCTA 265. CD33 AF5 HL .times. artificial aa
QVQLVQSGAEVKKPGASVKVSCKASGYTFTNYGMNWVKQAPGQGLKWMGWINTYTGEPTYADDFK
F12Q HL
GRVTMTSDTSTSTAYLELHNLRSDDTAVYYCARWSWSDGYYVYFDYWGQGTTVTVSSggggsg- gg
gsggggsDIVMTQSPDSLTVSLGERTTINCKSSQSVLDSSKNKNSLAWYQQKPGQPPKLLLSWAS
TRESGIPDRFSGSGSGTDFTLTIDSLQPEDSATYYCQQSAHFPITFGQGTRLEIKSGGGGSEVQL
VESGGGLVQPGGSLKLSCAASGFTFNSYAMNWVRQAPGKGLEWVARIRSKYNNYATYYADSVKGR
FTISRDDSKNTAYLQMNNLKTEDTAVYYCVRHGNFGNSYVSWWAYWGQGTLVTVSSGGGGSGGGG
SGGGGSQTVVTQEPSLTVSPGGTVTLTCGSSTGAVTSGNYPNWVQQKPGQAPRGLIGGTKFLAPG
TPARFSGSLLGGKAALTLSGVQPEDEAEYYCVLWYSNRWVFGGGTKLTVL 266. CD33 AF5 HL
.times. artificial nt
CAGGTGCAGCTGGTGCAGTCTGGAGCTGAGGTGAAGAAGCCTGGAGCGTCAGTCAAGGTCTCCTG
F12Q HL
CAAGGCTAGCGGGTATACCTTCACAAACTATGGAATGAACTGGGTGAAGCAGGCTCCAGGACA- GG
GTTTAAAGTGGATGGGCTGGATAAACACCTACACTGGAGAGCCAACATATGCTGATGACTTCAAG
GGACGGGTTACCATGACTTCGGATACCTCTACCAGCACTGCCTATTTGGAACTCCACAACCTCAG
AAGTGATGACACGGCTGTATATTACTGTGCGCGCTGGAGTTGGAGTGATGGTTACTACGTTTACT
TTGACTACTGGGGCCAAGGCACTACGGTCACCGTCTCCTCAGGTGGTGGTGGTTCTGGCGGCGGC
GGCTCCGGTGGTGGTGGTTCTGACATCGTGATGACACAGTCTCCAGACTCCCTGACTGTGTCTCT
GGGCGAGAGGACCACCATCAACTGCAAGTCCAGCCAGAGTGTTTTAGACAGCTCCAAGAATAAGA
ACTCCTTAGCTTGGTACCAGCAGAAACCAGGACAGCCTCCTAAATTACTCCTTTCCTGGGCATCT
ACGCGGGAATCCGGGATCCCTGACCGATTCAGTGGCAGCGGGTCTGGGACAGATTTCACTCTCAC
TATTGACAGCCTGCAGCCTGAAGATTCTGCAACTTACTATTGTCAACAGTCTGCCCACTTCCCGA
TCACCTTTGGCCAAGGGACACGACTGGAGATTAAATCCGGAGGTGGTGGCTCCGAGGTGCAGCTG
GTCGAGTCTGGAGGAGGATTGGTGCAGCCTGGAGGGTCATTGAAACTCTCATGTGCAGCCTCTGG
ATTCACCTTCAATAGCTACGCCATGAACTGGGTCCGCCAGGCTCCAGGAAAGGGTTTGGAATGGG
TTGCTCGCATAAGAAGTAAATATAATAATTATGCAACATATTATGCCGATTCAGTGAAAGGCAGG
TTCACCATCTCCAGAGATGATTCAAAAAACACTGCCTATCTACAAATGAACAACTTGAAAACTGA
GGACACTGCCGTGTACTACTGTGTGAGACATGGGAACTTCGGTAATAGCTACGTTTCCTGGTGGG
CTTACTGGGGCCAAGGGACTCTGGTCACCGTCTCCTCAGGTGGTGGTGGTTCTGGCGGCGGCGGC
TCCGGTGGTGGTGGTTCTCAGACTGTTGTGACTCAGGAACCTTCACTCACCGTATCACCTGGTGG
AACAGTCACACTCACTTGTGGCTCCTCGACTGGGGCTGTTACATCTGGCAACTACCCAAACTGGG
TCCAACAAAAACCAGGTCAGGCACCCCGTGGTCTAATAGGTGGGACTAAGTTCCTCGCCCCCGGT
ACTCCTGCCAGATTCTCAGGCTCCCTGCTTGGAGGCAAGGCTGCCCTCACCCTCTCAGGGGTACA
GCCAGAGGATGAGGCAGAATATTACTGTGTTCTATGGTACAGCAACCGCTGGGTGTTCGGTGGAG
GAACCAAACTGACTGTCCTA 267. CD33 AF5 HL .times. I2C artificial aa
QVQLVQSGAEVKKPGASVKVSCKASGYTFTNYGMNWVKQAPGQGLKWMGWINTYTGEPTYADDFK
HL
GRVTMTSDTSTSTAYLELHNLRSDDTAVYYCARWSWSDGYYVYFDYWGQGTTVTVSSggggsggg
gsggggsDIVMTQSPDSLTVSLGERTTINCKSSQSVLDSSKNKNSLAWYQQKPGQPPKLLLSWAS
TRESGIPDRFSGSGSGTDFTLTIDSLQPEDSATYYCQQSAHFPITFGQGTRLEIKSGGGGSEVQL
VESGGGLVQPGGSLKLSCAASGFTFNKYAMNWVRQAPGKGLEWVARIRSKYNNYATYYADSVKDR
FTISRDDSKNTAYLQMNNLKTEDTAVYYCVRHGNFGNSYISYWAYWGQGTLVTVSSGGGGSGGGG
SGGGGSQTVVTQEPSLTVSPGGTVTLTCGSSTGAVTSGNYPNWVQQKPGQAPRGLIGGTKFLAPG
TPARFSGSLLGGKAALTLSGVQPEDEAEYYCVLWYSNRWVFGGGTKLTVL 268. CD33 AF5 HL
.times. I2C artificial nt
CAGGTGCAGCTGGTGCAGTCTGGAGCTGAGGTGAAGAAGCCTGGAGCGTCAGTCAAGGTCTCCTG
HL
CAAGGCTAGCGGGTATACCTTCACAAACTATGGAATGAACTGGGTGAAGCAGGCTCCAGGACAGG
GTTTAAAGTGGATGGGCTGGATAAACACCTACACTGGAGAGCCAACATATGCTGATGACTTCAAG
GGACGGGTTACCATGACTTCGGATACCTCTACCAGCACTGCCTATTTGGAACTCCACAACCTCAG
AAGTGATGACACGGCTGTATATTACTGTGCGCGCTGGAGTTGGAGTGATGGTTACTACGTTTACT
TTGACTACTGGGGCCAAGGCACTACGGTCACCGTCTCCTCAGGTGGTGGTGGTTCTGGCGGCGGC
GGCTCCGGTGGTGGTGGTTCTGACATCGTGATGACACAGTCTCCAGACTCCCTGACTGTGTCTCT
GGGCGAGAGGACCACCATCAACTGCAAGTCCAGCCAGAGTGTTTTAGACAGCTCCAAGAATAAGA
ACTCCTTAGCTTGGTACCAGCAGAAACCAGGACAGCCTCCTAAATTACTCCTTTCCTGGGCATCT
ACGCGGGAATCCGGGATCCCTGACCGATTCAGTGGCAGCGGGTCTGGGACAGATTTCACTCTCAC
TATTGACAGCCTGCAGCCTGAAGATTCTGCAACTTACTATTGTCAACAGTCTGCCCACTTCCCGA
TCACCTTTGGCCAAGGGACACGACTGGAGATTAAATCCGGAGGTGGTGGCTCCGAGGTGCAGCTG
GTCGAGTCTGGAGGAGGATTGGTGCAGCCTGGAGGGTCATTGAAACTCTCATGTGCAGCCTCTGG
ATTCACCTTCAATAAGTACGCCATGAACTGGGTCCGCCAGGCTCCAGGAAAGGGTTTGGAATGGG
TTGCTCGCATAAGAAGTAAATATAATAATTATGCAACATATTATGCCGATTCAGTGAAAGACAGG
TTCACCATCTCCAGAGATGATTCAAAAAACACTGCCTATCTACAAATGAACAACTTGAAAACTGA
GGACACTGCCGTGTACTACTGTGTGAGACATGGGAACTTCGGTAATAGCTACATATCCTACTGGG
CTTACTGGGGCCAAGGGACTCTGGTCACCGTCTCCTCAGGTGGTGGTGGTTCTGGCGGCGGCGGC
TCCGGTGGTGGTGGTTCTCAGACTGTTGTGACTCAGGAACCTTCACTCACCGTATCACCTGGTGG
AACAGTCACACTCACTTGTGGCTCCTCGACTGGGGCTGTTACATCTGGCAACTACCCAAACTGGG
TCCAACAAAAACCAGGTCAGGCACCCCGTGGTCTAATAGGTGGGACTAAGTTCCTCGCCCCCGGT
ACTCCTGCCAGATTCTCAGGCTCCCTGCTTGGAGGCAAGGCTGCCCTCACCCTCTCAGGGGTACA
GCCAGAGGATGAGGCAGAATATTACTGTGTTCTATGGTACAGCAACCGCTGGGTGTTCGGTGGAG
GAACCAAACTGACTGTCCTA 269. CD33 AC8 HL .times. H2C artificial aa
QVQLVQSGAEVKKPGESVKVSCKASGYTFTNYGMNWVKQAPGQGLKWMGWINTYTGEPTYADDFK
HL
GRVTMTTDTSTSTAYMEIRNLRNDDTAVYYCARWSWSDGYYVYFDYWGQGTTVTVSSggggsggg
gsggggsDIVMTQSPDSLTVSLGERTTINCKSSQSVLDSSKNKNSLAWYQQKPGQPPKLLLSWAS
TRESGIPDRFSGSGSGTDFTLTIDSLQPEDSATYYCQQSAHFPITFGQGTRLEIKSGGGGSEVQL
VESGGGLVQPGGSLKLSCAASGFTFNKYAMNWVRQAPGKGLEWVARIRSKYNNYATYYADSVKDR
FTISRDDSKNTAYLQMNNLKTEDTAVYYCVRHGNFGNSYISYWAYWGQGTLVTVSSGGGGSGGGG
SGGGGSQTVVTQEPSLTVSPGGTVTLTCGSSTGAVTSGYYPNWVQQKPGQAPRGLIGGTKFLAPG
TPARFSGSLLGGKAALTLSGVQPEDEAEYYCALWYSNRWVFGGGTKLTVL 270. CD33 AC8 HL
.times. H2C artificial nt
CAGGTGCAGCTGGTGCAGTCTGGAGCTGAGGTGAAGAAGCCTGGAGAGTCAGTCAAGGTCTCCTG
HL
CAAGGCTAGCGGGTATACCTTCACAAACTATGGAATGAACTGGGTGAAGCAGGCTCCAGGACAGG
GTTTAAAGTGGATGGGCTGGATAAACACCTACACTGGAGAGCCAACATATGCTGATGACTTCAAG
GGACGGGTTACCATGACTACGGATACCTCTACCAGCACTGCCTATATGGAAATCCGCAACCTCAG
AAATGATGACACGGCTGTATATTACTGTGCGCGCTGGAGTTGGAGTGATGGTTACTACGTTTACT
TTGACTACTGGGGCCAAGGCACTACGGTCACCGTCTCCTCAGGTGGTGGTGGTTCTGGCGGCGGC
GGCTCCGGTGGTGGTGGTTCTGACATCGTGATGACACAGTCTCCAGACTCCCTGACTGTGTCTCT
GGGCGAGAGGACCACCATCAACTGCAAGTCCAGCCAGAGTGTTTTAGACAGCTCCAAGAATAAGA
ACTCCTTAGCTTGGTACCAGCAGAAACCAGGACAGCCTCCTAAATTACTCCTTTCCTGGGCATCT
ACGCGGGAATCCGGGATCCCTGACCGATTCAGTGGCAGCGGGTCTGGGACAGATTTCACTCTCAC
TATTGACAGCCTGCAGCCTGAAGATTCTGCAACTTACTATTGTCAACAGTCTGCCCACTTCCCGA
TCACCTTTGGCCAAGGGACACGACTGGAGATTAAATCCGGAGGTGGTGGCTCCGAGGTGCAGCTG
GTCGAGTCTGGAGGAGGATTGGTGCAGCCTGGAGGGTCATTGAAACTCTCATGTGCAGCCTCTGG
ATTCACCTTCAATAAGTACGCCATGAACTGGGTCCGCCAGGCTCCAGGAAAGGGTTTGGAATGGG
TTGCTCGCATAAGAAGTAAATATAATAATTATGCAACATATTATGCCGATTCAGTGAAAGACAGG
TTCACCATCTCCAGAGATGATTCAAAAAACACTGCCTATCTACAAATGAACAACTTGAAAACTGA
GGACACTGCCGTGTACTACTGTGTGAGACATGGGAACTTCGGTAATAGCTACATATCCTACTGGG
CTTACTGGGGCCAAGGGACTCTGGTCACCGTCTCCTCAGGTGGTGGTGGTTCTGGCGGCGGCGGC
TCCGGTGGTGGTGGTTCTCAGACTGTTGTGACTCAGGAACCTTCACTCACCGTATCACCTGGTGG
AACAGTCACACTCACTTGTGGCTCCTCGACTGGGGCTGTTACATCTGGCTACTACCCAAACTGGG
TCCAACAAAAACCAGGTCAGGCACCCCGTGGTCTAATAGGTGGGACTAAGTTCCTCGCCCCCGGT
ACTCCTGCCAGATTCTCAGGCTCCCTGCTTGGAGGCAAGGCTGCCCTCACCCTCTCAGGGCTACA
GCCAGAGGATGAGGCAGAATATTACTGTGCTCTATGGTACAGCAACCGCTGGGTGTTCGGTGGAG
GAACCAAACTGACTGTCCTA 271. CD33 AC8 HL .times. artificial aa
QVQLVQSGAEVKKPGESVKVSCKASGYTFTNYGMNWVKQAPGQGLKWMGWINTYTGEPTYADDFK
F12Q HL
GRVTMTTDTSTSTAYMEIRNLRNDDTAVYYCARWSWSDGYYVYFDYWGQGTTVTVSSggggsg- gg
gsggggsDIVMTQSPDSLTVSLGERTTINCKSSQSVLDSSKNKNSLAWYQQKPGQPPKLLLSWAS
TRESGIPDRFSGSGSGTDFTLTIDSLQPEDSATYYCQQSAHFPITFGQGTRLEIKSGGGGSEVQL
VESGGGLVQPGGSLKLSCAASGFTFNSYAMNWVRQAPGKGLEWVARIRSKYNNYATYYADSVKGR
FTISRDDSKNTAYLQMNNLKTEDTAVYYCVRHGNFGNSYVSWWAYWGQGTLVTVSSGGGGSGGGG
SGGGGSQTVVTQEPSLTVSPGGTVTLTCGSSTGAVTSGNYPNWVQQKPGQAPRGLIGGTKFLAPG
TPARFSGSLLGGKAALTLSGVQPEDEAEYYCVLWYSNRWVFGGGTKLTVL 272. CD33 AC8 HL
.times. artificial nt
CAGGTGCAGCTGGTGCAGTCTGGAGCTGAGGTGAAGAAGCCTGGAGAGTCAGTCAAGGTCTCCTG
F12Q HL
CAAGGCTAGCGGGTATACCTTCACAAACTATGGAATGAACTGGGTGAAGCAGGCTCCAGGACA- GG
GTTTAAAGTGGATGGGCTGGATAAACACCTACACTGGAGAGCCAACATATGCTGATGACTTCAAG
GGACGGGTTACCATGACTACGGATACCTCTACCAGCACTGCCTATATGGAAATCCGCAACCTCAG
AAATGATGACACGGCTGTATATTACTGTGCGCGCTGGAGTTGGAGTGATGGTTACTACGTTTACT
TTGACTACTGGGGCCAAGGCACTACGGTCACCGTCTCCTCAGGTGGTGGTGGTTCTGGCGGCGGC
GGCTCCGGTGGTGGTGGTTCTGACATCGTGATGACACAGTCTCCAGACTCCCTGACTGTGTCTCT
GGGCGAGAGGACCACCATCAACTGCAAGTCCAGCCAGAGTGTTTTAGACAGCTCCAAGAATAAGA
ACTCCTTAGCTTGGTACCAGCAGAAACCAGGACAGCCTCCTAAATTACTCCTTTCCTGGGCATCT
ACGCGGGAATCCGGGATCCCTGACCGATTCAGTGGCAGCGGGTCTGGGACAGATTTCACTCTCAC
TATTGACAGCCTGCAGCCTGAAGATTCTGCAACTTACTATTGTCAACAGTCTGCCCACTTCCCGA
TCACCTTTGGCCAAGGGACACGACTGGAGATTAAATCCGGAGGTGGTGGCTCCGAGGTGCAGCTG
GTCGAGTCTGGAGGAGGATTGGTGCAGCCTGGAGGGTCATTGAAACTCTCATGTGCAGCCTCTGG
ATTCACCTTCAATAGCTACGCCATGAACTGGGTCCGCCAGGCTCCAGGAAAGGGTTTGGAATGGG
TTGCTCGCATAAGAAGTAAATATAATAATTATGCAACATATTATGCCGATTCAGTGAAAGGCAGG
TTCACCATCTCCAGAGATGATTCAAAAAACACTGCCTATCTACAAATGAACAACTTGAAAACTGA
GGACACTGCCGTGTACTACTGTGTGAGACATGGGAACTTCGGTAATAGCTACGTTTCCTGGTGGG
CTTACTGGGGCCAAGGGACTCTGGTCACCGTCTCCTCAGGTGGTGGTGGTTCTGGCGGCGGCGGC
TCCGGTGGTGGTGGTTCTCAGACTGTTGTGACTCAGGAACCTTCACTCACCGTATCACCTGGTGG
AACAGTCACACTCACTTGTGGCTCCTCGACTGGGGCTGTTACATCTGGCAACTACCCAAACTGGG
TCCAACAAAAACCAGGTCAGGCACCCCGTGGTCTAATAGGTGGGACTAAGTTCCTCGCCCCCGGT
ACTCCTGCCAGATTCTCAGGCTCCCTGCTTGGAGGCAAGGCTGCCCTCACCCTCTCAGGGGTACA
GCCAGAGGATGAGGCAGAATATTACTGTGTTCTATGGTACAGCAACCGCTGGGTGTTCGGTGGAG
GAACCAAACTGACTGTCCTA 273. CD33 AC8 HL .times. I2C artificial aa
QVQLVQSGAEVKKPGESVKVSCKASGYTFTNYGMNWVKQAPGQGLKWMGWINTYTGEPTYADDFK
HL
GRVTMTTDTSTSTAYMEIRNLRNDDTAVYYCARWSWSDGYYVYFDYWGQGTTVTVSSggggsggg
gsggggsDIVMTQSPDSLTVSLGERTTINCKSSQSVLDSSKNKNSLAWYQQKPGQPPKLLLSWAS
TRESGIPDRFSGSGSGTDFTLTIDSLQPEDSATYYCQQSAHFPITFGQGTRLEIKSGGGGSEVQL
VESGGGLVQPGGSLKLSCAASGFTFNKYAMNWVRQAPGKGLEWVARIRSKYNNYATYYADSVKDR
FTISRDDSKNTAYLQMNNLKTEDTAVYYCVRHGNFGNSYISYWAYWGQGTLVTVSSGGGGSGGGG
SGGGGSQTVVTQEPSLTVSPGGTVTLTCGSSTGAVTSGNYPNWVQQKPGQAPRGLIGGTKFLAPG
TPARFSGSLLGGKAALTLSGVQPEDEAEYYCVLWYSNRWVFGGGTKLTVL 274. CD33 AC8 HL
.times. I2C artificial nt
CAGGTGCAGCTGGTGCAGTCTGGAGCTGAGGTGAAGAAGCCTGGAGAGTCAGTCAAGGTCTCCTG
HL
CAAGGCTAGCGGGTATACCTTCACAAACTATGGAATGAACTGGGTGAAGCAGGCTCCAGGACAGG
GTTTAAAGTGGATGGGCTGGATAAACACCTACACTGGAGAGCCAACATATGCTGATGACTTCAAG
GGACGGGTTACCATGACTACGGATACCTCTACCAGCACTGCCTATATGGAAATCCGCAACCTCAG
AAATGATGACACGGCTGTATATTACTGTGCGCGCTGGAGTTGGAGTGATGGTTACTACGTTTACT
TTGACTACTGGGGCCAAGGCACTACGGTCACCGTCTCCTCAGGTGGTGGTGGTTCTGGCGGCGGC
GGCTCCGGTGGTGGTGGTTCTGACATCGTGATGACACAGTCTCCAGACTCCCTGACTGTGTCTCT
GGGCGAGAGGACCACCATCAACTGCAAGTCCAGCCAGAGTGTTTTAGACAGCTCCAAGAATAAGA
ACTCCTTAGCTTGGTACCAGCAGAAACCAGGACAGCCTCCTAAATTACTCCTTTCCTGGGCATCT
ACGCGGGAATCCGGGATCCCTGACCGATTCAGTGGCAGCGGGTCTGGGACAGATTTCACTCTCAC
TATTGACAGCCTGCAGCCTGAAGATTCTGCAACTTACTATTGTCAACAGTCTGCCCACTTCCCGA
TCACCTTTGGCCAAGGGACACGACTGGAGATTAAATCCGGAGGTGGTGGCTCCGAGGTGCAGCTG
GTCGAGTCTGGAGGAGGATTGGTGCAGCCTGGAGGGTCATTGAAACTCTCATGTGCAGCCTCTGG
ATTCACCTTCAATAAGTACGCCATGAACTGGGTCCGCCAGGCTCCAGGAAAGGGTTTGGAATGGG
TTGCTCGCATAAGAAGTAAATATAATAATTATGCAACATATTATGCCGATTCAGTGAAAGACAGG
TTCACCATCTCCAGAGATGATTCAAAAAACACTGCCTATCTACAAATGAACAACTTGAAAACTGA
GGACACTGCCGTGTACTACTGTGTGAGACATGGGAACTTCGGTAATAGCTACATATCCTACTGGG
CTTACTGGGGCCAAGGGACTCTGGTCACCGTCTCCTCAGGTGGTGGTGGTTCTGGCGGCGGCGGC
TCCGGTGGTGGTGGTTCTCAGACTGTTGTGACTCAGGAACCTTCACTCACCGTATCACCTGGTGG
AACAGTCACACTCACTTGTGGCTCCTCGACTGGGGCTGTTACATCTGGCAACTACCCAAACTGGG
TCCAACAAAAACCAGGTCAGGCACCCCGTGGTCTAATAGGTGGGACTAAGTTCCTCGCCCCCGGT
ACTCCTGCCAGATTCTCAGGCTCCCTGCTTGGAGGCAAGGCTGCCCTCACCCTCTCAGGGGTACA
GCCAGAGGATGAGGCAGAATATTACTGTGTTCTATGGTACAGCAACCGCTGGGTGTTCGGTGGAG
GAACCAAACTGACTGTCCTA 275. CD33 AH11 HL .times. artificial aa
QVQLVQSGAEVKKPGESVKVSCKASGYTFTNYGMNWVKQAPGQGLKWMGWINTYTGEPTYADDFK
H2C HL
GRVTMTSDTSTSTAYMEISSLRSDDTAVYYCARWSWSDGYYVYFDYWGQGTTVTVSSggggsgg- g
gsggggsDIVMTQSPDSLTVSLGERTTINCKSSQSVLDSSKNKNSLAWYQQKPGQPPKLLLSWAS
TRESGIPDRFSGSGSGTDFTLTIDSLQPEDSATYYCQQSAHFPITFGQGTRLEIKSGGGGSEVQL
VESGGGLVQPGGSLKLSCAASGFTFNKYAMNWVRQAPGKGLEWVARIRSKYNNYATYYADSVKDR
FTISRDDSKNTAYLQMNNLKTEDTAVYYCVRHGNFGNSYISYWAYWGQGTLVTVSSGGGGSGGGG
SGGGGSQTVVTQEPSLTVSPGGTVTLTCGSSTGAVTSGYYPNWVQQKPGQAPRGLIGGTKFLAPG
TPARFSGSLLGGKAALTLSGVQPEDEAEYYCALWYSNRWVFGGGTKLTVL 276. CD33 AH11
HL .times. artificial nt
CAGGTGCAGCTGGTGCAGTCTGGAGCTGAGGTGAAGAAGCCTGGAGAGTCAGTCAAGGTCTCCTG
H2C HL
CAAGGCTAGCGGGTATACCTTCACAAACTATGGAATGAACTGGGTGAAGCAGGCTCCAGGACAG- G
GTTTAAAGTGGATGGGCTGGATAAACACCTACACTGGAGAGCCAACATATGCTGATGACTTCAAG
GGACGGGTTACCATGACTTCGGATACCTCTACCAGCACTGCCTATATGGAAATCAGCAGCCTCAG
AAGTGATGACACGGCTGTATATTACTGTGCGCGCTGGAGTTGGAGTGATGGTTACTACGTTTACT
TTGACTACTGGGGCCAAGGCACTACGGTCACCGTCTCCTCAGGTGGTGGTGGTTCTGGCGGCGGC
GGCTCCGGTGGTGGTGGTTCTGACATCGTGATGACACAGTCTCCAGACTCCCTGACTGTGTCTCT
GGGCGAGAGGACCACCATCAACTGCAAGTCCAGCCAGAGTGTTTTAGACAGCTCCAAGAATAAGA
ACTCCTTAGCTTGGTACCAGCAGAAACCAGGACAGCCTCCTAAATTACTCCTTTCCTGGGCATCT
ACGCGGGAATCCGGGATCCCTGACCGATTCAGTGGCAGCGGGTCTGGGACAGATTTCACTCTCAC
TATTGACAGCCTGCAGCCTGAAGATTCTGCAACTTACTATTGTCAACAGTCTGCCCACTTCCCGA
TCACCTTTGGCCAAGGGACACGACTGGAGATTAAATCCGGAGGTGGTGGCTCCGAGGTGCAGCTG
GTCGAGTCTGGAGGAGGATTGGTGCAGCCTGGAGGGTCATTGAAACTCTCATGTGCAGCCTCTGG
ATTCACCTTCAATAAGTACGCCATGAACTGGGTCCGCCAGGCTCCAGGAAAGGGTTTGGAATGGG
TTGCTCGCATAAGAAGTAAATATAATAATTATGCAACATATTATGCCGATTCAGTGAAAGACAGG
TTCACCATCTCCAGAGATGATTCAAAAAACACTGCCTATCTACAAATGAACAACTTGAAAACTGA
GGACACTGCCGTGTACTACTGTGTGAGACATGGGAACTTCGGTAATAGCTACATATCCTACTGGG
CTTACTGGGGCCAAGGGACTCTGGTCACCGTCTCCTCAGGTGGTGGTGGTTCTGGCGGCGGCGGC
TCCGGTGGTGGTGGTTCTCAGACTGTTGTGACTCAGGAACCTTCACTCACCGTATCACCTGGTGG
AACAGTCACACTCACTTGTGGCTCCTCGACTGGGGCTGTTACATCTGGCTACTACCCAAACTGGG
TCCAACAAAAACCAGGTCAGGCACCCCGTGGTCTAATAGGTGGGACTAAGTTCCTCGCCCCCGGT
ACTCCTGCCAGATTCTCAGGCTCCCTGCTTGGAGGCAAGGCTGCCCTCACCCTCTCAGGGGTACA
GCCAGAGGATGAGGCAGAATATTACTGTGCTCTATGGTACAGCAACCGCTGGGTGTTCGGTGGAG
GAACCAAACTGACTGTCCTA 277. CD33 AH11 HL .times. artificial aa
QVQLVQSGAEVKKPGESVKVSCKASGYTFTNYGMNWVKQAPGQGLKWMGWINTYTGEPTYADDFK
F12Q HL
GRVTMTSDTSTSTAYMEISSLRSDDTAVYYCARWSWSDGYYVYFDYWGQGTTVTVSSggggsg- gg
gsggggsDIVMTQSPDSLTVSLGERTTINCKSSQSVLDSSKNKNSLAWYQQKPGQPPKLLLSWAS
TRESGIPDRFSGSGSGTDFTLTIDSLQPEDSATYYCQQSAHFPITFGQGTRLEIKSGGGGSEVQL
VESGGGLVQPGGSLKLSCAASGFTFNSYAMNWVRQAPGKGLEWVARIRSKYNNYATYYADSVKGR
FTISRDDSKNTAYLQMNNLKTEDTAVYYCVRHGNFGNSYVSWWAYWGQGTLVTVSSGGGGSGGGG
SGGGGSQTVVTQEPSLTVSPGGTVTLTCGSSTGAVTSGNYPNWVQQKPGQAPRGLIGGTKFLAPG
TPARFSGSLLGGKAALTLSGVQPEDEAEYYCVLWYSNRWVFGGGTKLTVL 278. CD33 AH11
HL .times. artificial nt
CAGGTGCAGCTGGTGCAGTCTGGAGCTGAGGTGAAGAAGCCTGGAGAGTCAGTCAAGGTCTCCTG
F12Q HL
CAAGGCTAGCGGGTATACCTTCACAAACTATGGAATGAACTGGGTGAAGCAGGCTCCAGGACA- GG
GTTTAAAGTGGATGGGCTGGATAAACACCTACACTGGAGAGCCAACATATGCTGATGACTTCAAG
GGACGGGTTACCATGACTTCGGATACCTCTACCAGCACTGCCTATATGGAAATCAGCAGCCTCAG
AAGTGATGACACGGCTGTATATTACTGTGCGCGCTGGAGTTGGAGTGATGGTTACTACGTTTACT
TTGACTACTGGGGCCAAGGCACTACGGTCACCGTCTCCTCAGGTGGTGGTGGTTCTGGCGGCGGC
GGCTCCGGTGGTGGTGGTTCTGACATCGTGATGACACAGTCTCCAGACTCCCTGACTGTGTCTCT
GGGCGAGAGGACCACCATCAACTGCAAGTCCAGCCAGAGTGTTTTAGACAGCTCCAAGAATAAGA
ACTCCTTAGCTTGGTACCAGCAGAAACCAGGACAGCCTCCTAAATTACTCCTTTCCTGGGCATCT
ACGCGGGAATCCGGGATCCCTGACCGATTCAGTGGCAGCGGGTCTGGGACAGATTTCACTCTCAC
TATTGACAGCCTGCAGCCTGAAGATTCTGCAACTTACTATTGTCAACAGTCTGCCCACTTCCCGA
TCACCTTTGGCCAAGGGACACGACTGGAGATTAAATCCGGAGGTGGTGGCTCCGAGGTGCAGCTG
GTCGAGTCTGGAGGAGGATTGGTGCAGCCTGGAGGGTCATTGAAACTCTCATGTGCAGCCTCTGG
ATTCACCTTCAATAGCTACGCCATGAACTGGGTCCGCCAGGCTCCAGGAAAGGGTTTGGAATGGG
TTGCTCGCATAAGAAGTAAATATAATAATTATGCAACATATTATGCCGATTCAGTGAAAGGCAGG
TTCACCATCTCCAGAGATGATTCAAAAAACACTGCCTATCTACAAATGAACAACTTGAAAACTGA
GGACACTGCCGTGTACTACTGTGTGAGACATGGGAACTTCGGTAATAGCTACGTTTCCTGGTGGG
CTTACTGGGGCCAAGGGACTCTGGTCACCGTCTCCTCAGGTGGTGGTGGTTCTGGCGGCGGCGGC
TCCGGTGGTGGTGGTTCTCAGACTGTTGTGACTCAGGAACCTTCACTCACCGTATCACCTGGTGG
AACAGTCACACTCACTTGTGGCTCCTCGACTGGGGCTGTTACATCTGGCAACTACCCAAACTGGG
TCCAACAAAAACCAGGTCAGGCACCCCGTGGTCTAATAGGTGGGACTAAGTTCCTCGCCCCCGGT
ACTCCTGCCAGATTCTCAGGCTCCCTGCTTGGAGGCAAGGCTGCCCTCACCCTCTCAGGGGTACA
GCCAGAGGATGAGGCAGAATATTACTGTGTTCTATGGTACAGCAACCGCTGGGTGTTCGGTGGAG
GAACCAAACTGACTGTCCTA 279. CD33 AH11 HL .times. artificial aa
QVQLVQSGAEVKKPGESVKVSCKASGYTFTNYGMNWVKQAPGQGLKWMGWINTYTGEPTYADDFK
I2C HL
GRVTMTSDTSTSTAYMEISSLRSDDTAVYYCARWSWSDGYYVYFDYWGQGTTVTVSSggggsgg- g
gsggggsDIVMTQSPDSLTVSLGERTTINCKSSQSVLDSSKNKNSLAWYQQKPGQPPKLLLSWAS
TRESGIPDRFSGSGSGTDFTLTIDSLQPEDSATYYCQQSAHFPITFGQGTRLEIKSGGGGSEVQL
VESGGGLVQPGGSLKLSCAASGFTFNKYAMNWVRQAPGKGLEWVARIRSKYNNYATYYADSVKDR
FTISRDDSKNTAYLQMNNLKTEDTAVYYCVRHGNFGNSYISYWAYWGQGTLVTVSSGGGGSGGGG
SGGGGSQTVVTQEPSLTVSPGGTVTLTCGSSTGAVTSGNYPNWVQQKPGQAPRGLIGGTKFLAPG
TPARFSGSLLGGKAALTLSGVQPEDEAEYYCVLWYSNRWVFGGGTKLTVL 280. CD33 AH11
HL .times. artificial nt
CAGGTGCAGCTGGTGCAGTCTGGAGCTGAGGTGAAGAAGCCTGGAGAGTCAGTCAAGGTCTCCTG
I2C HL
CAAGGCTAGCGGGTATACCTTCACAAACTATGGAATGAACTGGGTGAAGCAGGCTCCAGGACAG- G
GTTTAAAGTGGATGGGCTGGATAAACACCTACACTGGAGAGCCAACATATGCTGATGACTTCAAG
GGACGGGTTACCATGACTTCGGATACCTCTACCAGCACTGCCTATATGGAAATCAGCAGCCTCAG
AAGTGATGACACGGCTGTATATTACTGTGCGCGCTGGAGTTGGAGTGATGGTTACTACGTTTACT
TTGACTACTGGGGCCAAGGCACTACGGTCACCGTCTCCTCAGGTGGTGGTGGTTCTGGCGGCGGC
GGCTCCGGTGGTGGTGGTTCTGACATCGTGATGACACAGTCTCCAGACTCCCTGACTGTGTCTCT
GGGCGAGAGGACCACCATCAACTGCAAGTCCAGCCAGAGTGTTTTAGACAGCTCCAAGAATAAGA
ACTCCTTAGCTTGGTACCAGCAGAAACCAGGACAGCCTCCTAAATTACTCCTTTCCTGGGCATCT
ACGCGGGAATCCGGGATCCCTGACCGATTCAGTGGCAGCGGGTCTGGGACAGATTTCACTCTCAC
TATTGACAGCCTGCAGCCTGAAGATTCTGCAACTTACTATTGTCAACAGTCTGCCCACTTCCCGA
TCACCTTTGGCCAAGGGACACGACTGGAGATTAAATCCGGAGGTGGTGGCTCCGAGGTGCAGCTG
GTCGAGTCTGGAGGAGGATTGGTGCAGCCTGGAGGGTCATTGAAACTCTCATGTGCAGCCTCTGG
ATTCACCTTCAATAAGTACGCCATGAACTGGGTCCGCCAGGCTCCAGGAAAGGGTTTGGAATGGG
TTGCTCGCATAAGAAGTAAATATAATAATTATGCAACATATTATGCCGATTCAGTGAAAGACAGG
TTCACCATCTCCAGAGATGATTCAAAAAACACTGCCTATCTACAAATGAACAACTTGAAAACTGA
GGACACTGCCGTGTACTACTGTGTGAGACATGGGAACTTCGGTAATAGCTACATATCCTACTGGG
CTTACTGGGGCCAAGGGACTCTGGTCACCGTCTCCTCAGGTGGTGGTGGTTCTGGCGGCGGCGGC
TCCGGTGGTGGTGGTTCTCAGACTGTTGTGACTCAGGAACCTTCACTCACCGTATCACCTGGTGG
AACAGTCACACTCACTTGTGGCTCCTCGACTGGGGCTGTTACATCTGGCAACTACCCAAACTGGG
TCCAACAAAAACCAGGTCAGGCACCCCGTGGTCTAATAGGTGGGACTAAGTTCCTCGCCCCCGGT
ACTCCTGCCAGATTCTCAGGCTCCCTGCTTGGAGGCAAGGCTGCCCTCACCCTCTCAGGGGTACA
GCCAGAGGATGAGGCAGAATATTACTGTGTTCTATGGTACAGCAACCGCTGGGTGTTCGGTGGAG
GAACCAAACTGACTGTCCTA 281. CD33 B3 HL .times. H2C artificial aa
QVQLVQSGAEVKKPGESVKVSCKASGYTFTNYGMNWVKQAPGQGLEWMGWINTYTGETNYADKFQ
HL
GRVTFTSDTSTSTAYMELRNLKSDDTAVYYCARWSWSDGYYVYFDYWGQGTTVTVSSggggsggg
gsggggsDIVMTQSPDSMTVSLGERTTINCKSSQSVLDSSTNKNSLAWYQQKPGQPPKLLLSWAS
TRESGIPDRFSGSGSGTDFTLTIDSLQPEDSATYYCQQSAHFPITFGQGTRLDIKSGGGGSEVQL
VESGGGLVQPGGSLKLSCAASGFTFNKYAMNWVRQAPGKGLEWVARIRSKYNNYATYYADSVKDR
FTISRDDSKNTAYLQMNNLKTEDTAVYYCVRHGNFGNSYISYWAYWGQGTLVTVSSGGGGSGGGG
SGGGGSQTVVTQEPSLTVSPGGTVTLTCGSSTGAVTSGYYPNWVQQKPGQAPRGLIGGTKFLAPG
TPARFSGSLLGGKAALTLSGVQPEDEAEYYCALWYSNRWVFGGGTKLTVL 282. CD33 B3 HL
.times. H2C artificial nt
CAGGTGCAGCTGGTGCAGTCTGGAGCTGAGGTGAAGAAGCCTGGAGAGTCAGTCAAGGTCTCCTG
HL
CAAGGCTAGCGGGTATACCTTCACAAACTATGGAATGAACTGGGTGAAGCAGGCTCCAGGACAGG
GTTTAGAGTGGATGGGCTGGATAAACACCTACACTGGAGAGACAAACTATGCTGATAAGTTCCAG
GGACGCGTTACCTTCACTTCGGATACCTCTACCAGCACTGCCTATATGGAACTCCGCAACCTCAA
AAGTGATGACACGGCTGTATATTACTGTGCGCGCTGGAGTTGGAGTGATGGTTACTACGTTTACT
TTGACTACTGGGGCCAAGGCACTACGGTCACCGTCTCCTCAGGTGGTGGTGGTTCTGGCGGCGGC
GGCTCCGGTGGTGGTGGTTCTGACATCGTGATGACACAGTCTCCAGACTCCATGACTGTGTCTCT
GGGCGAGAGGACCACCATCAACTGCAAGTCCAGCCAGAGTGTTTTAGACAGCTCCACGAATAAGA
ACTCCTTAGCTTGGTACCAGCAGAAACCAGGACAGCCTCCTAAATTACTCCTTTCCTGGGCATCT
ACGCGGGAATCCGGGATCCCTGACCGATTCAGTGGCAGCGGGTCTGGGACAGATTTCACTCTCAC
TATTGACAGCCTGCAGCCTGAAGATTCTGCAACTTACTATTGTCAACAGTCTGCCCACTTCCCGA
TCACCTTTGGCCAAGGGACACGACTGGACATTAAATCCGGAGGTGGTGGCTCCGAGGTGCAGCTG
GTCGAGTCTGGAGGAGGATTGGTGCAGCCTGGAGGGTCATTGAAACTCTCATGTGCAGCCTCTGG
ATTCACCTTCAATAAGTACGCCATGAACTGGGTCCGCCAGGCTCCAGGAAAGGGTTTGGAATGGG
TTGCTCGCATAAGAAGTAAATATAATAATTATGCAACATATTATGCCGATTCAGTGAAAGACAGG
TTCACCATCTCCAGAGATGATTCAAAAAACACTGCCTATCTACAAATGAACAACTTGAAAACTGA
GGACACTGCCGTGTACTACTGTGTGAGACATGGGAACTTCGGTAATAGCTACATATCCTACTGGG
CTTACTGGGGCCAAGGGACTCTGGTCACCGTCTCCTCAGGTGGTGGTGGTTCTGGCGGCGGCGGC
TCCGGTGGTGGTGGTTCTCAGACTGTTGTGACTCAGGAACCTTCACTCACCGTATCACCTGGTGG
AACAGTCACACTCACTTGTGGCTCCTCGACTGGGGCTGTTACATCTGGCTACTACCCAAACTGGG
TCCAACAAAAACCAGGTCAGGCACCCCGTGGTCTAATAGGTGGGACTAAGTTCCTCGCCCCCGGT
ACTCCTGCCAGATTCTCAGGCTCCCTGCTTGGAGGCAAGGCTGCCCTCACCCTCTCAGGGGTACA
GCCAGAGGATGAGGCAGAATATTACTGTGCTCTATGGTACAGCAACCGCTGGGTGTTCGGTGGAG
GAACCAAACTGACTGTCCTA 283. CD33 B3 HL .times. F12Q artificial aa
QVQLVQSGAEVKKPGESVKVSCKASGYTFTNYGMNWVKQAPGQGLEWMGWINTYTGETNYADKFQ
HL
GRVTFTSDTSTSTAYMELRNLKSDDTAVYYCARWSWSDGYYVYFDYWGQGTTVTVSSggggsggg
gsggggsDIVMTQSPDSMTVSLGERTTINCKSSQSVLDSSTNKNSLAWYQQKPGQPPKLLLSWAS
TRESGIPDRFSGSGSGTDFTLTIDSLQPEDSATYYCQQSAHFPITFGQGTRLDIKSGGGGSEVQL
VESGGGLVQPGGSLKLSCAASGFTFNSYAMNWVRQAPGKGLEWVARIRSKYNNYATYYADSVKGR
FTISRDDSKNTAYLQMNNLKTEDTAVYYCVRHGNFGNSYVSWWAYWGQGTLVTVSSGGGGSGGGG
SGGGGSQTVVTQEPSLTVSPGGTVTLTCGSSTGAVTSGNYPNWVQQKPGQAPRGLIGGTKFLAPG
TPARFSGSLLGGKAALTLSGVQPEDEAEYYCVLWYSNRWVFGGGTKLTVL 284. CD33 B3 HL
.times. F12Q artificial nt
CAGGTGCAGCTGGTGCAGTCTGGAGCTGAGGTGAAGAAGCCTGGAGAGTCAGTCAAGGTCTCCTG
HL
CAAGGCTAGCGGGTATACCTTCACAAACTATGGAATGAACTGGGTGAAGCAGGCTCCAGGACAGG
GTTTAGAGTGGATGGGCTGGATAAACACCTACACTGGAGAGACAAACTATGCTGATAAGTTCCAG
GGACGCGTTACCTTCACTTCGGATACCTCTACCAGCACTGCCTATATGGAACTCCGCAACCTCAA
AAGTGATGACACGGCTGTATATTACTGTGCGCGCTGGAGTTGGAGTGATGGTTACTACGTTTACT
TTGACTACTGGGGCCAAGGCACTACGGTCACCGTCTCCTCAGGTGGTGGTGGTTCTGGCGGCGGC
GGCTCCGGTGGTGGTGGTTCTGACATCGTGATGACACAGTCTCCAGACTCCATGACTGTGTCTCT
GGGCGAGAGGACCACCATCAACTGCAAGTCCAGCCAGAGTGTTTTAGACAGCTCCACGAATAAGA
ACTCCTTAGCTTGGTACCAGCAGAAACCAGGACAGCCTCCTAAATTACTCCTTTCCTGGGCATCT
ACGCGGGAATCCGGGATCCCTGACCGATTCAGTGGCAGCGGGTCTGGGACAGATTTCACTCTCAC
TATTGACAGCCTGCAGCCTGAAGATTCTGCAACTTACTATTGTCAACAGTCTGCCCACTTCCCGA
TCACCTTTGGCCAAGGGACACGACTGGACATTAAATCCGGAGGTGGTGGCTCCGAGGTGCAGCTG
GTCGAGTCTGGAGGAGGATTGGTGCAGCCTGGAGGGTCATTGAAACTCTCATGTGCAGCCTCTGG
ATTCACCTTCAATAGCTACGCCATGAACTGGGTCCGCCAGGCTCCAGGAAAGGGTTTGGAATGGG
TTGCTCGCATAAGAAGTAAATATAATAATTATGCAACATATTATGCCGATTCAGTGAAAGGCAGG
TTCACCATCTCCAGAGATGATTCAAAAAACACTGCCTATCTACAAATGAACAACTTGAAAACTGA
GGACACTGCCGTGTACTACTGTGTGAGACATGGGAACTTCGGTAATAGCTACGTTTCCTGGTGGG
CTTACTGGGGCCAAGGGACTCTGGTCACCGTCTCCTCAGGTGGTGGTGGTTCTGGCGGCGGCGGC
TCCGGTGGTGGTGGTTCTCAGACTGTTGTGACTCAGGAACCTTCACTCACCGTATCACCTGGTGG
AACAGTCACACTCACTTGTGGCTCCTCGACTGGGGCTGTTACATCTGGCAACTACCCAAACTGGG
TCCAACAAAAACCAGGTCAGGCACCCCGTGGTCTAATAGGTGGGACTAAGTTCCTCGCCCCCGGT
ACTCCTGCCAGATTCTCAGGCTCCCTGCTTGGAGGCAAGGCTGCCCTCACCCTCTCAGGGGTACA
GCCAGAGGATGAGGCAGAATATTACTGTGTTCTATGGTACAGCAACCGCTGGGTGTTCGGTGGAG
GAACCAAACTGACTGTCCTA 285. CD33 B3 HL .times. I2C artificial aa
QVQLVQSGAEVKKPGESVKVSCKASGYTFTNYGMNWVKQAPGQGLEWMGWINTYTGETNYADKFQ
HL
GRVTFTSDTSTSTAYMELRNLKSDDTAVYYCARWSWSDGYYVYFDYWGQGTTVTVSSggggsggg
gsggggsDIVMTQSPDSMTVSLGERTTINCKSSQSVLDSSTNKNSLAWYQQKPGQPPKLLLSWAS
TRESGIPDRFSGSGSGTDFTLTIDSLQPEDSATYYCQQSAHFPITFGQGTRLDIKSGGGGSEVQL
VESGGGLVQPGGSLKLSCAASGFTFNKYAMNWVRQAPGKGLEWVARIRSKYNNYATYYADSVKDR
FTISRDDSKNTAYLQMNNLKTEDTAVYYCVRHGNFGNSYISYWAYWGQGTLVTVSSGGGGSGGGG
SGGGGSQTVVTQEPSLTVSPGGTVTLTCGSSTGAVTSGNYPNWVQQKPGQAPRGLIGGTKFLAPG
TPARFSGSLLGGKAALTLSGVQPEDEAEYYCVLWYSNRWVFGGGTKLTVL 286. CD33 B3 HL
.times. I2C artificial nt
CAGGTGCAGCTGGTGCAGTCTGGAGCTGAGGTGAAGAAGCCTGGAGAGTCAGTCAAGGTCTCCTG
HL
CAAGGCTAGCGGGTATACCTTCACAAACTATGGAATGAACTGGGTGAAGCAGGCTCCAGGACAGG
GTTTAGAGTGGATGGGCTGGATAAACACCTACACTGGAGAGACAAACTATGCTGATAAGTTCCAG
GGACGCGTTACCTTCACTTCGGATACCTCTACCAGCACTGCCTATATGGAACTCCGCAACCTCAA
AAGTGATGACACGGCTGTATATTACTGTGCGCGCTGGAGTTGGAGTGATGGTTACTACGTTTACT
TTGACTACTGGGGCCAAGGCACTACGGTCACCGTCTCCTCAGGTGGTGGTGGTTCTGGCGGCGGC
GGCTCCGGTGGTGGTGGTTCTGACATCGTGATGACACAGTCTCCAGACTCCATGACTGTGTCTCT
GGGCGAGAGGACCACCATCAACTGCAAGTCCAGCCAGAGTGTTTTAGACAGCTCCACGAATAAGA
ACTCCTTAGCTTGGTACCAGCAGAAACCAGGACAGCCTCCTAAATTACTCCTTTCCTGGGCATCT
ACGCGGGAATCCGGGATCCCTGACCGATTCAGTGGCAGCGGGTCTGGGACAGATTTCACTCTCAC
TATTGACAGCCTGCAGCCTGAAGATTCTGCAACTTACTATTGTCAACAGTCTGCCCACTTCCCGA
TCACCTTTGGCCAAGGGACACGACTGGACATTAAATCCGGAGGTGGTGGCTCCGAGGTGCAGCTG
GTCGAGTCTGGAGGAGGATTGGTGCAGCCTGGAGGGTCATTGAAACTCTCATGTGCAGCCTCTGG
ATTCACCTTCAATAAGTACGCCATGAACTGGGTCCGCCAGGCTCCAGGAAAGGGTTTGGAATGGG
TTGCTCGCATAAGAAGTAAATATAATAATTATGCAACATATTATGCCGATTCAGTGAAAGACAGG
TTCACCATCTCCAGAGATGATTCAAAAAACACTGCCTATCTACAAATGAACAACTTGAAAACTGA
GGACACTGCCGTGTACTACTGTGTGAGACATGGGAACTTCGGTAATAGCTACATATCCTACTGGG
CTTACTGGGGCCAAGGGACTCTGGTCACCGTCTCCTCAGGTGGTGGTGGTTCTGGCGGCGGCGGC
TCCGGTGGTGGTGGTTCTCAGACTGTTGTGACTCAGGAACCTTCACTCACCGTATCACCTGGTGG
AACAGTCACACTCACTTGTGGCTCCTCGACTGGGGCTGTTACATCTGGCAACTACCCAAACTGGG
TCCAACAAAAACCAGGTCAGGCACCCCGTGGTCTAATAGGTGGGACTAAGTTCCTCGCCCCCGGT
ACTCCTGCCAGATTCTCAGGCTCCCTGCTTGGAGGCAAGGCTGCCCTCACCCTCTCAGGGGTACA
GCCAGAGGATGAGGCAGAATATTACTGTGTTCTATGGTACAGCAACCGCTGGGTGTTCGGTGGAG
GAACCAAACTGACTGTCCTA 287. CD33 F2 HL .times. H2C artificial aa
QVQLVQSGAEVKKPGESVKVSCKASGYTFTNYGMNWVKQAPGQGLEWMGWINTYTGETNYADKFQ
HL
GRVTFTSDTSTSTAYMELRNLKSDDTAVYYCARWSWSDGYYVYFDYWGQGTTVTVSSggggsggg
gsggggsDIVMTQSPDSLSVSLGERTTINCKSSQSVLDSSTNKNSLAWYQQKPGQPPKLLLSWAS
TRESGIPDRFSGSGSGTDFTLTIDSLQPEDSATYYCQQSAHFPITFGQGTRLEIKSGGGGSEVQL
VESGGGLVQPGGSLKLSCAASGFTFNKYAMNWVRQAPGKGLEWVARIRSKYNNYATYYADSVKDR
FTISRDDSKNTAYLQMNNLKTEDTAVYYCVRHGNFGNSYISYWAYWGQGTLVTVSSGGGGSGGGG
SGGGGSQTVVTQEPSLTVSPGGTVTLTCGSSTGAVTSGYYPNWVQQKPGQAPRGLIGGTKFLAPG
TPARFSGSLLGGKAALTLSGVQPEDEAEYYCALWYSNRWVFGGGTKLTVL 288. CD33 F2 HL
.times. H2C artificial nt
CAGGTGCAGCTGGTGCAGTCTGGAGCTGAGGTGAAGAAGCCTGGAGAGTCAGTCAAGGTCTCCTG
HL
CAAGGCTAGCGGGTATACCTTCACAAACTATGGAATGAACTGGGTGAAGCAGGCTCCAGGACAGG
GTTTAGAGTGGATGGGCTGGATAAACACCTACACTGGAGAGACAAACTATGCTGATAAGTTCCAG
GGACGCGTTACCTTCACTTCGGATACCTCTACCAGCACTGCCTATATGGAACTCCGCAACCTCAA
AAGTGATGACACGGCTGTATATTACTGTGCGCGCTGGAGTTGGAGTGATGGTTACTACGTTTACT
TTGACTACTGGGGCCAAGGCACTACGGTCACCGTCTCCTCAGGTGGTGGTGGTTCTGGCGGCGGC
GGCTCCGGTGGTGGTGGTTCTGACATCGTGATGACACAGTCTCCAGACTCCCTGTCTGTGTCTCT
GGGCGAGAGGACCACCATCAACTGCAAGTCCAGCCAGAGTGTTTTAGACAGCTCCACGAATAAGA
ACTCCTTAGCTTGGTACCAGCAGAAACCAGGACAGCCTCCTAAATTACTCCTTTCCTGGGCATCT
ACGCGGGAATCCGGGATCCCTGACCGATTCAGTGGCAGCGGGTCTGGGACAGATTTCACTCTCAC
TATTGACAGCCTGCAGCCTGAAGATTCTGCAACTTACTATTGTCAACAGTCTGCCCACTTCCCGA
TCACCTTTGGCCAAGGGACACGACTGGAGATTAAATCCGGAGGTGGTGGCTCCGAGGTGCAGCTG
GTCGAGTCTGGAGGAGGATTGGTGCAGCCTGGAGGGTCATTGAAACTCTCATGTGCAGCCTCTGG
ATTCACCTTCAATAAGTACGCCATGAACTGGGTCCGCCAGGCTCCAGGAAAGGGTTTGGAATGGG
TTGCTCGCATAAGAAGTAAATATAATAATTATGCAACATATTATGCCGATTCAGTGAAAGACAGG
TTCACCATCTCCAGAGATGATTCAAAAAACACTGCCTATCTACAAATGAACAACTTGAAAACTGA
GGACACTGCCGTGTACTACTGTGTGAGACATGGGAACTTCGGTAATAGCTACATATCCTACTGGG
CTTACTGGGGCCAAGGGACTCTGGTCACCGTCTCCTCAGGTGGTGGTGGTTCTGGCGGCGGCGGC
TCCGGTGGTGGTGGTTCTCAGACTGTTGTGACTCAGGAACCTTCACTCACCGTATCACCTGGTGG
AACAGTCACACTCACTTGTGGCTCCTCGACTGGGGCTGTTACATCTGGCTACTACCCAAACTGGG
TCCAACAAAAACCAGGTCAGGCACCCCGTGGTCTAATAGGTGGGACTAAGTTCCTCGCCCCCGGT
ACTCCTGCCAGATTCTCAGGCTCCCTGCTTGGAGGCAAGGCTGCCCTCACCCTCTCAGGGGTACA
GCCAGAGGATGAGGCAGAATATTACTGTGCTCTATGGTACAGCAACCGCTGGGTGTTCGGTGGAG
GAACCAAACTGACTGTCCTA 289. CD33 F2 HL .times. F12Q artificial aa
QVQLVQSGAEVKKPGESVKVSCKASGYTFTNYGMNWVKQAPGQGLEWMGWINTYTGETNYADKFQ
HL
GRVTFTSDTSTSTAYMELRNLKSDDTAVYYCARWSWSDGYYVYFDYWGQGTTVTVSSggggsggg
gsggggsDIVMTQSPDSLSVSLGERTTINCKSSQSVLDSSTNKNSLAWYQQKPGQPPKLLLSWAS
TRESGIPDRFSGSGSGTDFTLTIDSLQPEDSATYYCQQSAHFPITFGQGTRLEIKSGGGGSEVQL
VESGGGLVQPGGSLKLSCAASGFTFNSYAMNWVRQAPGKGLEWVARIRSKYNNYATYYADSVKGR
FTISRDDSKNTAYLQMNNLKTEDTAVYYCVRHGNFGNSYVSWWAYWGQGTLVTVSSGGGGSGGGG
SGGGGSQTVVTQEPSLTVSPGGTVTLTCGSSTGAVTSGNYPNWVQQKPGQAPRGLIGGTKFLAPG
TPARFSGSLLGGKAALTLSGVQPEDEAEYYCVLWYSNRWVFGGGTKLTVL 290. CD33 F2 HL
.times. F12Q artificial nt
CAGGTGCAGCTGGTGCAGTCTGGAGCTGAGGTGAAGAAGCCTGGAGAGTCAGTCAAGGTCTCCTG
HL
CAAGGCTAGCGGGTATACCTTCACAAACTATGGAATGAACTGGGTGAAGCAGGCTCCAGGACAGG
GTTTAGAGTGGATGGGCTGGATAAACACCTACACTGGAGAGACAAACTATGCTGATAAGTTCCAG
GGACGCGTTACCTTCACTTCGGATACCTCTACCAGCACTGCCTATATGGAACTCCGCAACCTCAA
AAGTGATGACACGGCTGTATATTACTGTGCGCGCTGGAGTTGGAGTGATGGTTACTACGTTTACT
TTGACTACTGGGGCCAAGGCACTACGGTCACCGTCTCCTCAGGTGGTGGTGGTTCTGGCGGCGGC
GGCTCCGGTGGTGGTGGTTCTGACATCGTGATGACACAGTCTCCAGACTCCCTGTCTGTGTCTCT
GGGCGAGAGGACCACCATCAACTGCAAGTCCAGCCAGAGTGTTTTAGACAGCTCCACGAATAAGA
ACTCCTTAGCTTGGTACCAGCAGAAACCAGGACAGCCTCCTAAATTACTCCTTTCCTGGGCATCT
ACGCGGGAATCCGGGATCCCTGACCGATTCAGTGGCAGCGGGTCTGGGACAGATTTCACTCTCAC
TATTGACAGCCTGCAGCCTGAAGATTCTGCAACTTACTATTGTCAACAGTCTGCCCACTTCCCGA
TCACCTTTGGCCAAGGGACACGACTGGAGATTAAATCCGGAGGTGGTGGCTCCGAGGTGCAGCTG
GTCGAGTCTGGAGGAGGATTGGTGCAGCCTGGAGGGTCATTGAAACTCTCATGTGCAGCCTCTGG
ATTCACCTTCAATAGCTACGCCATGAACTGGGTCCGCCAGGCTCCAGGAAAGGGTTTGGAATGGG
TTGCTCGCATAAGAAGTAAATATAATAATTATGCAACATATTATGCCGATTCAGTGAAAGGCAGG
TTCACCATCTCCAGAGATGATTCAAAAAACACTGCCTATCTACAAATGAACAACTTGAAAACTGA
GGACACTGCCGTGTACTACTGTGTGAGACATGGGAACTTCGGTAATAGCTACGTTTCCTGGTGGG
CTTACTGGGGCCAAGGGACTCTGGTCACCGTCTCCTCAGGTGGTGGTGGTTCTGGCGGCGGCGGC
TCCGGTGGTGGTGGTTCTCAGACTGTTGTGACTCAGGAACCTTCACTCACCGTATCACCTGGTGG
AACAGTCACACTCACTTGTGGCTCCTCGACTGGGGCTGTTACATCTGGCAACTACCCAAACTGGG
TCCAACAAAAACCAGGTCAGGCACCCCGTGGTCTAATAGGTGGGACTAAGTTCCTCGCCCCCGGT
ACTCCTGCCAGATTCTCAGGCTCCCTGCTTGGAGGCAAGGCTGCCCTCACCCTCTCAGGGGTACA
GCCAGAGGATGAGGCAGAATATTACTGTGTTCTATGGTACAGCAACCGCTGGGTGTTCGGTGGAG
GAACCAAACTGACTGTCCTA 291. CD33 F2 HL .times. I2C artificial aa
QVQLVQSGAEVKKPGESVKVSCKASGYTFTNYGMNWVKQAPGQGLEWMGWINTYTGETNYADKFQ
HL
GRVTFTSDTSTSTAYMELRNLKSDDTAVYYCARWSWSDGYYVYFDYWGQGTTVTVSSggggsggg
gsggggsDIVMTQSPDSLSVSLGERTTINCKSSQSVLDSSTNKNSLAWYQQKPGQPPKLLLSWAS
TRESGIPDRFSGSGSGTDFTLTIDSLQPEDSATYYCQQSAHFPITFGQGTRLEIKSGGGGSEVQL
VESGGGLVQPGGSLKLSCAASGFTFNKYAMNWVRQAPGKGLEWVARIRSKYNNYATYYADSVKDR
FTISRDDSKNTAYLQMNNLKTEDTAVYYCVRHGNFGNSYISYWAYWGQGTLVTVSSGGGGSGGGG
SGGGGSQTVVTQEPSLTVSPGGTVTLTCGSSTGAVTSGNYPNWVQQKPGQAPRGLIGGTKFLAPG
TPARFSGSLLGGKAALTLSGVQPEDEAEYYCVLWYSNRWVFGGGTKLTVL 292. CD33 F2 HL
.times. I2C artificial nt
CAGGTGCAGCTGGTGCAGTCTGGAGCTGAGGTGAAGAAGCCTGGAGAGTCAGTCAAGGTCTCCTG
HL
CAAGGCTAGCGGGTATACCTTCACAAACTATGGAATGAACTGGGTGAAGCAGGCTCCAGGACAGG
GTTTAGAGTGGATGGGCTGGATAAACACCTACACTGGAGAGACAAACTATGCTGATAAGTTCCAG
GGACGCGTTACCTTCACTTCGGATACCTCTACCAGCACTGCCTATATGGAACTCCGCAACCTCAA
AAGTGATGACACGGCTGTATATTACTGTGCGCGCTGGAGTTGGAGTGATGGTTACTACGTTTACT
TTGACTACTGGGGCCAAGGCACTACGGTCACCGTCTCCTCAGGTGGTGGTGGTTCTGGCGGCGGC
GGCTCCGGTGGTGGTGGTTCTGACATCGTGATGACACAGTCTCCAGACTCCCTGTCTGTGTCTCT
GGGCGAGAGGACCACCATCAACTGCAAGTCCAGCCAGAGTGTTTTAGACAGCTCCACGAATAAGA
ACTCCTTAGCTTGGTACCAGCAGAAACCAGGACAGCCTCCTAAATTACTCCTTTCCTGGGCATCT
ACGCGGGAATCCGGGATCCCTGACCGATTCAGTGGCAGCGGGTCTGGGACAGATTTCACTCTCAC
TATTGACAGCCTGCAGCCTGAAGATTCTGCAACTTACTATTGTCAACAGTCTGCCCACTTCCCGA
TCACCTTTGGCCAAGGGACACGACTGGAGATTAAATCCGGAGGTGGTGGCTCCGAGGTGCAGCTG
GTCGAGTCTGGAGGAGGATTGGTGCAGCCTGGAGGGTCATTGAAACTCTCATGTGCAGCCTCTGG
ATTCACCTTCAATAAGTACGCCATGAACTGGGTCCGCCAGGCTCCAGGAAAGGGTTTGGAATGGG
TTGCTCGCATAAGAAGTAAATATAATAATTATGCAACATATTATGCCGATTCAGTGAAAGACAGG
TTCACCATCTCCAGAGATGATTCAAAAAACACTGCCTATCTACAAATGAACAACTTGAAAACTGA
GGACACTGCCGTGTACTACTGTGTGAGACATGGGAACTTCGGTAATAGCTACATATCCTACTGGG
CTTACTGGGGCCAAGGGACTCTGGTCACCGTCTCCTCAGGTGGTGGTGGTTCTGGCGGCGGCGGC
TCCGGTGGTGGTGGTTCTCAGACTGTTGTGACTCAGGAACCTTCACTCACCGTATCACCTGGTGG
AACAGTCACACTCACTTGTGGCTCCTCGACTGGGGCTGTTACATCTGGCAACTACCCAAACTGGG
TCCAACAAAAACCAGGTCAGGCACCCCGTGGTCTAATAGGTGGGACTAAGTTCCTCGCCCCCGGT
ACTCCTGCCAGATTCTCAGGCTCCCTGCTTGGAGGCAAGGCTGCCCTCACCCTCTCAGGGGTACA
GCCAGAGGATGAGGCAGAATATTACTGTGTTCTATGGTACAGCAACCGCTGGGTGTTCGGTGGAG
GAACCAAACTGACTGTCCTA 293. CD33 B10 HL .times. H2C artificial aa
QVQLVQSGAEVKKPGESVKVSCKASGYTFTNYGMNWVKQAPGQGLEWMGWINTYTGEPTYADKFQ
HL
GRVTMTTDTSTSTAYMEIRNLRSDDTAVYYCARWSWSDGYYVYFDYWGQGTTVTVSSggggsggg
gsggggsDIVMTQSPDSLTVSLGERTTINCKSSQSVLDSSNNKNSLAWYQQKPGQPPKLLLSWAS
TRESGIPDRFSGSGSGTDFTLTIDGLQPEDSATYYCQQSAHFPITFGQGTRLEIKSGGGGSEVQL
VESGGGLVQPGGSLKLSCAASGFTFNKYAMNWVRQAPGKGLEWVARIRSKYNNYATYYADSVKDR
FTISRDDSKNTAYLQMNNLKTEDTAVYYCVRHGNFGNSYISYWAYWGQGTLVTVSSGGGGSGGGG
SGGGGSQTVVTQEPSLTVSPGGTVTLTCGSSTGAVTSGYYPNWVQQKPGQAPRGLIGGTKFLAPG
TPARFSGSLLGGKAALTLSGVQPEDEAEYYCALWYSNRWVFGGGTKLTVL 294. CD33 B10 HL
.times. H2C artificial nt
CAGGTGCAGCTGGTGCAGTCTGGAGCTGAGGTGAAGAAGCCTGGTGAGTCAGTCAAGGTCTCCTG
HL
CAAGGCTAGCGGGTATACCTTCACAAACTATGGAATGAACTGGGTGAAGCAGGCTCCAGGACAGG
GTTTAGAGTGGATGGGCTGGATAAACACCTACACTGGAGAGCCAACCTATGCTGATAAGTTCCAG
GGACGCGTTACCATGACTACGGATACCTCTACCAGCACTGCCTATATGGAAATCCGCAACCTCAG
AAGTGATGACACGGCTGTATATTACTGTGCGCGCTGGAGTTGGAGTGATGGTTACTACGTTTACT
TTGACTACTGGGGCCAAGGCACTACGGTCACCGTCTCCTCAGGTGGTGGTGGTTCTGGCGGCGGC
GGCTCCGGTGGTGGTGGTTCTGACATCGTGATGACACAGTCTCCAGACTCCCTGACTGTGTCTCT
GGGCGAGAGGACCACCATCAACTGCAAGTCCAGCCAGAGTGTTTTAGACAGCTCCAACAATAAGA
ACTCCTTAGCTTGGTACCAGCAGAAACCAGGACAGCCTCCTAAATTACTCCTTTCCTGGGCATCT
ACGCGGGAATCCGGGATCCCTGACCGATTCAGTGGCAGCGGTTCTGGGACAGATTTCACTCTCAC
TATTGACGGCCTGCAGCCTGAAGATTCTGCAACTTACTATTGTCAACAGTCTGCCCACTTCCCGA
TCACCTTTGGCCAAGGGACACGACTGGAGATTAAATCCGGAGGTGGTGGCTCCGAGGTGCAGCTG
GTCGAGTCTGGAGGAGGATTGGTGCAGCCTGGAGGGTCATTGAAACTCTCATGTGCAGCCTCTGG
ATTCACCTTCAATAAGTACGCCATGAACTGGGTCCGCCAGGCTCCAGGAAAGGGTTTGGAATGGG
TTGCTCGCATAAGAAGTAAATATAATAATTATGCAACATATTATGCCGATTCAGTGAAAGACAGG
TTCACCATCTCCAGAGATGATTCAAAAAACACTGCCTATCTACAAATGAACAACTTGAAAACTGA
GGACACTGCCGTGTACTACTGTGTGAGACATGGGAACTTCGGTAATAGCTACATATCCTACTGGG
CTTACTGGGGCCAAGGGACTCTGGTCACCGTCTCCTCAGGTGGTGGTGGTTCTGGCGGCGGCGGC
TCCGGTGGTGGTGGTTCTCAGACTGTTGTGACTCAGGAACCTTCACTCACCGTATCACCTGGTGG
AACAGTCACACTCACTTGTGGCTCCTCGACTGGGGCTGTTACATCTGGCTACTACCCAAACTGGG
TCCAACAAAAACCAGGTCAGGCACCCCGTGGTCTAATAGGTGGGACTAAGTTCCTCGCCCCCGGT
ACTCCTGCCAGATTCTCAGGCTCCCTGCTTGGAGGCAAGGCTGCCCTCACCCTCTCAGGGGTACA
GCCAGAGGATGAGGCAGAATATTACTGTGCTCTATGGTACAGCAACCGCTGGGTGTTCGGTGGAG
GAACCAAACTGACTGTCCTA 295. CD33 B10 HL .times. artificial aa
QVQLVQSGAEVKKPGESVKVSCKASGYTFTNYGMNWVKQAPGQGLEWMGWINTYTGEPTYADKFQ
F12Q HL
GRVTMTTDTSTSTAYMEIRNLRSDDTAVYYCARWSWSDGYYVYFDYWGQGTTVTVSSggggsg- gg
gsggggsDIVMTQSPDSLTVSLGERTTINCKSSQSVLDSSNNKNSLAWYQQKPGQPPKLLLSWAS
TRESGIPDRFSGSGSGTDFTLTIDGLQPEDSATYYCQQSAHFPITFGQGTRLEIKSGGGGSEVQL
VESGGGLVQPGGSLKLSCAASGFTFNSYAMNWVRQAPGKGLEWVARIRSKYNNYATYYADSVKGR
FTISRDDSKNTAYLQMNNLKTEDTAVYYCVRHGNFGNSYVSWWAYWGQGTLVTVSSGGGGSGGGG
SGGGGSQTVVTQEPSLTVSPGGTVTLTCGSSTGAVTSGNYPNWVQQKPGQAPRGLIGGTKFLAPG
TPARFSGSLLGGKAALTLSGVQPEDEAEYYCVLWYSNRWVFGGGTKLTVL 296. CD33 B10 HL
.times. artificial nt
CAGGTGCAGCTGGTGCAGTCTGGAGCTGAGGTGAAGAAGCCTGGTGAGTCAGTCAAGGTCTCCTG
F12Q HL
CAAGGCTAGCGGGTATACCTTCACAAACTATGGAATGAACTGGGTGAAGCAGGCTCCAGGACA- GG
GTTTAGAGTGGATGGGCTGGATAAACACCTACACTGGAGAGCCAACCTATGCTGATAAGTTCCAG
GGACGCGTTACCATGACTACGGATACCTCTACCAGCACTGCCTATATGGAAATCCGCAACCTCAG
AAGTGATGACACGGCTGTATATTACTGTGCGCGCTGGAGTTGGAGTGATGGTTACTACGTTTACT
TTGACTACTGGGGCCAAGGCACTACGGTCACCGTCTCCTCAGGTGGTGGTGGTTCTGGCGGCGGC
GGCTCCGGTGGTGGTGGTTCTGACATCGTGATGACACAGTCTCCAGACTCCCTGACTGTGTCTCT
GGGCGAGAGGACCACCATCAACTGCAAGTCCAGCCAGAGTGTTTTAGACAGCTCCAACAATAAGA
ACTCCTTAGCTTGGTACCAGCAGAAACCAGGACAGCCTCCTAAATTACTCCTTTCCTGGGCATCT
ACGCGGGAATCCGGGATCCCTGACCGATTCAGTGGCAGCGGTTCTGGGACAGATTTCACTCTCAC
TATTGACGGCCTGCAGCCTGAAGATTCTGCAACTTACTATTGTCAACAGTCTGCCCACTTCCCGA
TCACCTTTGGCCAAGGGACACGACTGGAGATTAAATCCGGAGGTGGTGGCTCCGAGGTGCAGCTG
GTCGAGTCTGGAGGAGGATTGGTGCAGCCTGGAGGGTCATTGAAACTCTCATGTGCAGCCTCTGG
ATTCACCTTCAATAGCTACGCCATGAACTGGGTCCGCCAGGCTCCAGGAAAGGGTTTGGAATGGG
TTGCTCGCATAAGAAGTAAATATAATAATTATGCAACATATTATGCCGATTCAGTGAAAGGCAGG
TTCACCATCTCCAGAGATGATTCAAAAAACACTGCCTATCTACAAATGAACAACTTGAAAACTGA
GGACACTGCCGTGTACTACTGTGTGAGACATGGGAACTTCGGTAATAGCTACGTTTCCTGGTGGG
CTTACTGGGGCCAAGGGACTCTGGTCACCGTCTCCTCAGGTGGTGGTGGTTCTGGCGGCGGCGGC
TCCGGTGGTGGTGGTTCTCAGACTGTTGTGACTCAGGAACCTTCACTCACCGTATCACCTGGTGG
AACAGTCACACTCACTTGTGGCTCCTCGACTGGGGCTGTTACATCTGGCAACTACCCAAACTGGG
TCCAACAAAAACCAGGTCAGGCACCCCGTGGTCTAATAGGTGGGACTAAGTTCCTCGCCCCCGGT
ACTCCTGCCAGATTCTCAGGCTCCCTGCTTGGAGGCAAGGCTGCCCTCACCCTCTCAGGGGTACA
GCCAGAGGATGAGGCAGAATATTACTGTGTTCTATGGTACAGCAACCGCTGGGTGTTCGGTGGAG
GAACCAAACTGACTGTCCTA 297. CD33 B10 HL .times. I2C artificial aa
QVQLVQSGAEVKKPGESVKVSCKASGYTFTNYGMNWVKQAPGQGLEWMGWINTYTGEPTYADKFQ
HL
GRVTMTTDTSTSTAYMEIRNLRSDDTAVYYCARWSWSDGYYVYFDYWGQGTTVTVSSggggsggg
gsggggsDIVMTQSPDSLTVSLGERTTINCKSSQSVLDSSNNKNSLAWYQQKPGQPPKLLLSWAS
TRESGIPDRFSGSGSGTDFTLTIDGLQPEDSATYYCQQSAHFPITFGQGTRLEIKSGGGGSEVQL
VESGGGLVQPGGSLKLSCAASGFTFNKYAMNWVRQAPGKGLEWVARIRSKYNNYATYYADSVKDR
FTISRDDSKNTAYLQMNNLKTEDTAVYYCVRHGNFGNSYISYWAYWGQGTLVTVSSGGGGSGGGG
SGGGGSQTVVTQEPSLTVSPGGTVTLTCGSSTGAVTSGNYPNWVQQKPGQAPRGLIGGTKFLAPG
TPARFSGSLLGGKAALTLSGVQPEDEAEYYCVLWYSNRWVFGGGTKLTVL 298. CD33 B10 HL
.times. I2C artificial nt
CAGGTGCAGCTGGTGCAGTCTGGAGCTGAGGTGAAGAAGCCTGGTGAGTCAGTCAAGGTCTCCTG
HL
CAAGGCTAGCGGGTATACCTTCACAAACTATGGAATGAACTGGGTGAAGCAGGCTCCAGGACAGG
GTTTAGAGTGGATGGGCTGGATAAACACCTACACTGGAGAGCCAACCTATGCTGATAAGTTCCAG
GGACGCGTTACCATGACTACGGATACCTCTACCAGCACTGCCTATATGGAAATCCGCAACCTCAG
AAGTGATGACACGGCTGTATATTACTGTGCGCGCTGGAGTTGGAGTGATGGTTACTACGTTTACT
TTGACTACTGGGGCCAAGGCACTACGGTCACCGTCTCCTCAGGTGGTGGTGGTTCTGGCGGCGGC
GGCTCCGGTGGTGGTGGTTCTGACATCGTGATGACACAGTCTCCAGACTCCCTGACTGTGTCTCT
GGGCGAGAGGACCACCATCAACTGCAAGTCCAGCCAGAGTGTTTTAGACAGCTCCAACAATAAGA
ACTCCTTAGCTTGGTACCAGCAGAAACCAGGACAGCCTCCTAAATTACTCCTTTCCTGGGCATCT
ACGCGGGAATCCGGGATCCCTGACCGATTCAGTGGCAGCGGTTCTGGGACAGATTTCACTCTCAC
TATTGACGGCCTGCAGCCTGAAGATTCTGCAACTTACTATTGTCAACAGTCTGCCCACTTCCCGA
TCACCTTTGGCCAAGGGACACGACTGGAGATTAAATCCGGAGGTGGTGGCTCCGAGGTGCAGCTG
GTCGAGTCTGGAGGAGGATTGGTGCAGCCTGGAGGGTCATTGAAACTCTCATGTGCAGCCTCTGG
ATTCACCTTCAATAAGTACGCCATGAACTGGGTCCGCCAGGCTCCAGGAAAGGGTTTGGAATGGG
TTGCTCGCATAAGAAGTAAATATAATAATTATGCAACATATTATGCCGATTCAGTGAAAGACAGG
TTCACCATCTCCAGAGATGATTCAAAAAACACTGCCTATCTACAAATGAACAACTTGAAAACTGA
GGACACTGCCGTGTACTACTGTGTGAGACATGGGAACTTCGGTAATAGCTACATATCCTACTGGG
CTTACTGGGGCCAAGGGACTCTGGTCACCGTCTCCTCAGGTGGTGGTGGTTCTGGCGGCGGCGGC
TCCGGTGGTGGTGGTTCTCAGACTGTTGTGACTCAGGAACCTTCACTCACCGTATCACCTGGTGG
AACAGTCACACTCACTTGTGGCTCCTCGACTGGGGCTGTTACATCTGGCAACTACCCAAACTGGG
TCCAACAAAAACCAGGTCAGGCACCCCGTGGTCTAATAGGTGGGACTAAGTTCCTCGCCCCCGGT
ACTCCTGCCAGATTCTCAGGCTCCCTGCTTGGAGGCAAGGCTGCCCTCACCCTCTCAGGGGTACA
GCCAGAGGATGAGGCAGAATATTACTGTGTTCTATGGTACAGCAACCGCTGGGTGTTCGGTGGAG
GAACCAAACTGACTGTCCTA 299. CD33 E11 HL .times. H2C artificial aa
QVQLVQSGAEVKKPGESVKVSCKASGYTFTNYGMNWVKQAPGQGLEWMGWINTYTGEPTYADKFQ
HL
GRVTMTTDTSTSTAYMEIRNLGGDDTAVYYCARWSWSDGYYVYFDYWGQGTSVTVSSggggsggg
gsggggsDIVMTQSPDSLTVSLGERTTINCKSSQSVLDSSTNKNSLAWYQQKPGQPPKLLLSWAS
TRESGIPDRFSGSGSGTDFTLTIDSPQPEDSATYYCQQSAHFPITFGQGTRLEIKSGGGGSEVQL
VESGGGLVQPGGSLKLSCAASGFTFNKYAMNWVRQAPGKGLEWVARIRSKYNNYATYYADSVKDR
FTISRDDSKNTAYLQMNNLKTEDTAVYYCVRHGNFGNSYISYWAYWGQGTLVTVSSGGGGSGGGG
SGGGGSQTVVTQEPSLTVSPGGTVTLTCGSSTGAVTSGYYPNWVQQKPGQAPRGLIGGTKFLAPG
TPARFSGSLLGGKAALTLSGVQPEDEAEYYCALWYSNRWVFGGGTKLTVL 300. CD33 E11 HL
.times. H2C artificial nt
CAGGTGCAGCTGGTGCAGTCTGGAGCTGAGGTGAAGAAGCCTGGAGAGTCAGTCAAGGTCTCCTG
HL
CAAGGCTAGCGGGTATACCTTCACAAACTATGGAATGAACTGGGTGAAGCAGGCTCCAGGACAGG
GTTTAGAGTGGATGGGCTGGATAAACACCTACACTGGAGAGCCAACCTATGCTGATAAGTTCCAG
GGACGCGTTACCATGACTACGGATACCTCTACCAGCACTGCCTATATGGAAATCCGCAACCTCGG
AGGTGATGACACGGCTGTATATTACTGTGCGCGCTGGAGTTGGAGTGATGGTTACTACGTTTACT
TTGACTACTGGGGCCAAGGCACTTCGGTCACCGTCTCCTCAGGTGGTGGTGGTTCTGGCGGCGGC
GGCTCCGGTGGTGGTGGTTCTGACATCGTGATGACACAGTCTCCAGACTCCCTGACTGTGTCTCT
GGGCGAGAGGACCACCATCAACTGCAAGTCCAGCCAGAGTGTTTTAGACAGCTCCACGAATAAGA
ACTCCTTAGCTTGGTACCAGCAGAAACCAGGACAGCCTCCTAAATTACTCCTTTCCTGGGCATCT
ACGCGGGAATCCGGGATCCCTGACCGATTCAGTGGCAGCGGGTCTGGGACAGATTTCACTCTCAC
TATTGACAGCCCGCAGCCTGAAGATTCTGCAACTTACTATTGTCAACAGTCTGCCCACTTCCCGA
TCACCTTTGGCCAAGGGACACGACTGGAGATTAAATCCGGAGGTGGTGGCTCCGAGGTGCAGCTG
GTCGAGTCTGGAGGAGGATTGGTGCAGCCTGGAGGGTCATTGAAACTCTCATGTGCAGCCTCTGG
ATTCACCTTCAATAAGTACGCCATGAACTGGGTCCGCCAGGCTCCAGGAAAGGGTTTGGAATGGG
TTGCTCGCATAAGAAGTAAATATAATAATTATGCAACATATTATGCCGATTCAGTGAAAGACAGG
TTCACCATCTCCAGAGATGATTCAAAAAACACTGCCTATCTACAAATGAACAACTTGAAAACTGA
GGACACTGCCGTGTACTACTGTGTGAGACATGGGAACTTCGGTAATAGCTACATATCCTACTGGG
CTTACTGGGGCCAAGGGACTCTGGTCACCGTCTCCTCAGGTGGTGGTGGTTCTGGCGGCGGCGGC
TCCGGTGGTGGTGGTTCTCAGACTGTTGTGACTCAGGAACCTTCACTCACCGTATCACCTGGTGG
AACAGTCACACTCACTTGTGGCTCCTCGACTGGGGCTGTTACATCTGGCTACTACCCAAACTGGG
TCCAACAAAAACCAGGTCAGGCACCCCGTGGTCTAATAGGTGGGACTAAGTTCCTCGCCCCCGGT
ACTCCTGCCAGATTCTCAGGCTCCCTGCTTGGAGGCAAGGCTGCCCTCACCCTCTCAGGGGTACA
GCCAGAGGATGAGGCAGAATATTACTGTGCTCTATGGTACAGCAACCGCTGGGTGTTCGGTGGAG
GAACCAAACTGACTGTCCTA 301. CD33 E11 HL .times. artificial aa
QVQLVQSGAEVKKPGESVKVSCKASGYTFTNYGMNWVKQAPGQGLEWMGWINTYTGEPTYADKFQ
F12Q HL
GRVTMTTDTSTSTAYMEIRNLGGDDTAVYYCARWSWSDGYYVYFDYWGQGTSVTVSSggggsg- gg
gsggggsDIVMTQSPDSLTVSLGERTTINCKSSQSVLDSSTNKNSLAWYQQKPGQPPKLLLSWAS
TRESGIPDRFSGSGSGTDFTLTIDSPQPEDSATYYCQQSAHFPITFGQGTRLEIKSGGGGSEVQL
VESGGGLVQPGGSLKLSCAASGFTFNSYAMNWVRQAPGKGLEWVARIRSKYNNYATYYADSVKGR
FTISRDDSKNTAYLQMNNLKTEDTAVYYCVRHGNFGNSYVSWWAYWGQGTLVTVSSGGGGSGGGG
SGGGGSQTVVTQEPSLTVSPGGTVTLTCGSSTGAVTSGNYPNWVQQKPGQAPRGLIGGTKFLAPG
TPARFSGSLLGGKAALTLSGVQPEDEAEYYCVLWYSNRWVFGGGTKLTVL 302. CD33 E11 HL
.times. artificial nt
CAGGTGCAGCTGGTGCAGTCTGGAGCTGAGGTGAAGAAGCCTGGAGAGTCAGTCAAGGTCTCCTG
F12Q HL
CAAGGCTAGCGGGTATACCTTCACAAACTATGGAATGAACTGGGTGAAGCAGGCTCCAGGACA- GG
GTTTAGAGTGGATGGGCTGGATAAACACCTACACTGGAGAGCCAACCTATGCTGATAAGTTCCAG
GGACGCGTTACCATGACTACGGATACCTCTACCAGCACTGCCTATATGGAAATCCGCAACCTCGG
AGGTGATGACACGGCTGTATATTACTGTGCGCGCTGGAGTTGGAGTGATGGTTACTACGTTTACT
TTGACTACTGGGGCCAAGGCACTTCGGTCACCGTCTCCTCAGGTGGTGGTGGTTCTGGCGGCGGC
GGCTCCGGTGGTGGTGGTTCTGACATCGTGATGACACAGTCTCCAGACTCCCTGACTGTGTCTCT
GGGCGAGAGGACCACCATCAACTGCAAGTCCAGCCAGAGTGTTTTAGACAGCTCCACGAATAAGA
ACTCCTTAGCTTGGTACCAGCAGAAACCAGGACAGCCTCCTAAATTACTCCTTTCCTGGGCATCT
ACGCGGGAATCCGGGATCCCTGACCGATTCAGTGGCAGCGGGTCTGGGACAGATTTCACTCTCAC
TATTGACAGCCCGCAGCCTGAAGATTCTGCAACTTACTATTGTCAACAGTCTGCCCACTTCCCGA
TCACCTTTGGCCAAGGGACACGACTGGAGATTAAATCCGGAGGTGGTGGCTCCGAGGTGCAGCTG
GTCGAGTCTGGAGGAGGATTGGTGCAGCCTGGAGGGTCATTGAAACTCTCATGTGCAGCCTCTGG
ATTCACCTTCAATAGCTACGCCATGAACTGGGTCCGCCAGGCTCCAGGAAAGGGTTTGGAATGGG
TTGCTCGCATAAGAAGTAAATATAATAATTATGCAACATATTATGCCGATTCAGTGAAAGGCAGG
TTCACCATCTCCAGAGATGATTCAAAAAACACTGCCTATCTACAAATGAACAACTTGAAAACTGA
GGACACTGCCGTGTACTACTGTGTGAGACATGGGAACTTCGGTAATAGCTACGTTTCCTGGTGGG
CTTACTGGGGCCAAGGGACTCTGGTCACCGTCTCCTCAGGTGGTGGTGGTTCTGGCGGCGGCGGC
TCCGGTGGTGGTGGTTCTCAGACTGTTGTGACTCAGGAACCTTCACTCACCGTATCACCTGGTGG
AACAGTCACACTCACTTGTGGCTCCTCGACTGGGGCTGTTACATCTGGCAACTACCCAAACTGGG
TCCAACAAAAACCAGGTCAGGCACCCCGTGGTCTAATAGGTGGGACTAAGTTCCTCGCCCCCGGT
ACTCCTGCCAGATTCTCAGGCTCCCTGCTTGGAGGCAAGGCTGCCCTCACCCTCTCAGGGGTACA
GCCAGAGGATGAGGCAGAATATTACTGTGTTCTATGGTACAGCAACCGCTGGGTGTTCGGTGGAG
GAACCAAACTGACTGTCCTA 303. CD33 E11 HL .times. I2C artificial aa
QVQLVQSGAEVKKPGESVKVSCKASGYTFTNYGMNWVKQAPGQGLEWMGWINTYTGEPTYADKFQ
HL
GRVTMTTDTSTSTAYMEIRNLGGDDTAVYYCARWSWSDGYYVYFDYWGQGTSVTVSSggggsggg
gsggggsDIVMTQSPDSLTVSLGERTTINCKSSQSVLDSSTNKNSLAWYQQKPGQPPKLLLSWAS
TRESGIPDRFSGSGSGTDFTLTIDSPQPEDSATYYCQQSAHFPITFGQGTRLEIKSGGGGSEVQL
VESGGGLVQPGGSLKLSCAASGFTFNKYAMNWVRQAPGKGLEWVARIRSKYNNYATYYADSVKDR
FTISRDDSKNTAYLQMNNLKTEDTAVYYCVRHGNFGNSYISYWAYWGQGTLVTVSSGGGGSGGGG
SGGGGSQTVVTQEPSLTVSPGGTVTLTCGSSTGAVTSGNYPNWVQQKPGQAPRGLIGGTKFLAPG
TPARFSGSLLGGKAALTLSGVQPEDEAEYYCVLWYSNRWVFGGGTKLTVL 304. CD33 E11 HL
.times. I2C artificial nt
CAGGTGCAGCTGGTGCAGTCTGGAGCTGAGGTGAAGAAGCCTGGAGAGTCAGTCAAGGTCTCCTG
HL
CAAGGCTAGCGGGTATACCTTCACAAACTATGGAATGAACTGGGTGAAGCAGGCTCCAGGACAGG
GTTTAGAGTGGATGGGCTGGATAAACACCTACACTGGAGAGCCAACCTATGCTGATAAGTTCCAG
GGACGCGTTACCATGACTACGGATACCTCTACCAGCACTGCCTATATGGAAATCCGCAACCTCGG
AGGTGATGACACGGCTGTATATTACTGTGCGCGCTGGAGTTGGAGTGATGGTTACTACGTTTACT
TTGACTACTGGGGCCAAGGCACTTCGGTCACCGTCTCCTCAGGTGGTGGTGGTTCTGGCGGCGGC
GGCTCCGGTGGTGGTGGTTCTGACATCGTGATGACACAGTCTCCAGACTCCCTGACTGTGTCTCT
GGGCGAGAGGACCACCATCAACTGCAAGTCCAGCCAGAGTGTTTTAGACAGCTCCACGAATAAGA
ACTCCTTAGCTTGGTACCAGCAGAAACCAGGACAGCCTCCTAAATTACTCCTTTCCTGGGCATCT
ACGCGGGAATCCGGGATCCCTGACCGATTCAGTGGCAGCGGGTCTGGGACAGATTTCACTCTCAC
TATTGACAGCCCGCAGCCTGAAGATTCTGCAACTTACTATTGTCAACAGTCTGCCCACTTCCCGA
TCACCTTTGGCCAAGGGACACGACTGGAGATTAAATCCGGAGGTGGTGGCTCCGAGGTGCAGCTG
GTCGAGTCTGGAGGAGGATTGGTGCAGCCTGGAGGGTCATTGAAACTCTCATGTGCAGCCTCTGG
ATTCACCTTCAATAAGTACGCCATGAACTGGGTCCGCCAGGCTCCAGGAAAGGGTTTGGAATGGG
TTGCTCGCATAAGAAGTAAATATAATAATTATGCAACATATTATGCCGATTCAGTGAAAGACAGG
TTCACCATCTCCAGAGATGATTCAAAAAACACTGCCTATCTACAAATGAACAACTTGAAAACTGA
GGACACTGCCGTGTACTACTGTGTGAGACATGGGAACTTCGGTAATAGCTACATATCCTACTGGG
CTTACTGGGGCCAAGGGACTCTGGTCACCGTCTCCTCAGGTGGTGGTGGTTCTGGCGGCGGCGGC
TCCGGTGGTGGTGGTTCTCAGACTGTTGTGACTCAGGAACCTTCACTCACCGTATCACCTGGTGG
AACAGTCACACTCACTTGTGGCTCCTCGACTGGGGCTGTTACATCTGGCAACTACCCAAACTGGG
TCCAACAAAAACCAGGTCAGGCACCCCGTGGTCTAATAGGTGGGACTAAGTTCCTCGCCCCCGGT
ACTCCTGCCAGATTCTCAGGCTCCCTGCTTGGAGGCAAGGCTGCCCTCACCCTCTCAGGGGTACA
GCCAGAGGATGAGGCAGAATATTACTGTGTTCTATGGTACAGCAACCGCTGGGTGTTCGGTGGAG
GAACCAAACTGACTGTCCTA 305. CD33 human nt
ATGGGATGGAGCTGTATCATCCTCTTCTTGGTAGCAACAGCTACAGGTGTACACTCCGATCCAAA
TTTCTGGCTGCAAGTGCAGGAGTCAGTGACGGTACAGGAGGGTTTGTGCGTCCTCGTGCCCTGCA
CTTTCTTCCATCCCATACCCTACTACGACAAGAACTCCCCAGTTCATGGTTACTGGTTCCGGGAA
GGAGCCATTATATCCGGGGACTCTCCAGTGGCCACAAACAAGCTAGATCAAGAAGTACAGGAGGA
GACTCAGGGCAGATTCCGCCTCCTTGGGGATCCCAGTAGGAACAACTGCTCCCTGAGCATCGTAG
ACGCCAGGAGGAGGGATAATGGTTCATACTTCTTTCGGATGGAGAGAGGAAGTACCAAATACAGT
TACAAATCTCCCCAGCTCTCTGTGCATGTGACAGACTTGACCCACAGGCCCAAAATCCTCATCCC
TGGCACTCTAGAACCCGGCCACTCCAAAAACCTGACCTGCTCTGTGTCCTGGGCCTGTGAGCAGG
GAACACCCCCGATCTTCTCCTGGTTGTCAGCTGCCCCCACCTCCCTGGGCCCCAGGACTACTCAC
TCCTCGGTGCTCATAATCACCCCACGGCCCCAGGACCACGGCACCAACCTGACCTGTCAGGTGAA
GTTCGCTGGAGCTGGTGTGACTACGGAGAGAACCATCCAGCTCAACGTCACCTATGTTCCACAGA
ACCCAACAACTGGTATCTTTCCAGGAGATGGCTCAGGGAAACAAGAGACCAGAGCAGGAGTGGTT
CATGGGGCCATTGGAGGAGCTGGTGTTACAGCCCTGCTCGCTCTTTGTCTCTGCCTCATCTTCTT
CATAGTGAAGACCCACAGGAGGAAAGCAGCCAGGACAGCAGTGGGCAGGAATGACACCCACCCTA
CCACAGGGTCAGCCTCCCCGAAACACCAGAAGAAGTCCAAGTTACATGGCCCCACTGAAACCTCA
AGCTGTTCAGGTGCCGCCCCTACTGTGGAGATGGATGAGGAGCTGCATTATGCTTCCCTCAACTT
TCATGGGATGAATCCTTCCAAGGACACCTCCACCGAATACTCAGAGGTCAGGACCCAGTCCGGGC
ATCATCACCATCATCATTGA 306. CD33 human aa
MGWSCIILFLVATATGVHSDPNFWLQVQESVTVQEGLCVLVPCTFFHPIPYYDKNSPVHGYWFRE
GAIISGDSPVATNKLDQEVQEETQGRFRLLGDPSRNNCSLSIVDARRRDNGSYFFRMERGSTKYS
YKSPQLSVHVTDLTHRPKILIPGTLEPGHSKNLTCSVSWACEQGTPPIFSWLSAAPTSLGPRTTH
SSVLIITPRPQDHGTNLTCQVKFAGAGVTTERTIQLNVTYVPQNPTTGIFPGDGSGKQETRAGVV
HGAIGGAGVTALLALCLCLIFFIVKTHRRKAARTAVGRNDTHPTTGSASPKHQKKSKLHGPTETS
SCSGAAPTVEMDEELHYASLNFHGMNPSKDTSTEYSEVRTQSGHHHHHH 307. CD33 macaque
nt
ATGCCGCTGCTGCTACTGCTGCCCCTGCTGTGGGCAGGGGCCCTGGCTATGGATCCAAGAGTCAG
GCTGGAAGTGCAGGAGTCAGTGACAGTACAGGAGGGTTTGTGCGTCCTTGTGCCCTGCACTTTCT
TCCATCCCGTACCCTACCACACCAGGAATTCCCCAGTTCATGGTTACTGGTTCCGGGAAGGAGCC
ATTGTATCCTTGGACTCTCCAGTGGCCACAAACAAGCTAGATCAAGAAGTACAGGAGGAGACCCA
GGGCCGATTCCGCCTCCTTGGGGATCCCAGTAGGAACAACTGCTCCCTGAGCATCGTAGATGCCA
GGAGGAGGGATAACGGTTCATACTTCTTTCGGATGGAGAAAGGAAGTACCAAATACAGTTACAAA
TCTACCCAGCTCTCTGTGCATGTGACAGACTTGACCCACAGGCCCCAAATCCTCATCCCTGGAGC
CCTAGACCCTGACCACTCCAAAAACCTGACCTGCTCTGTGCCCTGGGCCTGTGAGCAGGGAACAC
CTCCAATCTTCTCCTGGATGTCAGCTGCCCCCACCTCCCTGGGCCTCAGGACCACTCACTCCTCG
GTGCTCATAATCACCCCACGGCCCCAGGACCACGGCACCAACCTCACCTGTCAGGTGAAGTTCCC
TGGAGCTGGCGTGACCACGGAGAGAACCATCCAGCTCAATGTCTCCTATGCTTCACAGAACCCAA
GAACTGATATCTTTCTAGGAGACGGCTCAGGGAAACAAGGAGTGGTTCAGGGAGCCATCGGGGGA
GCTGGTGTCACAGTCCTGCTCGCTCTTTGTCTCTGCCTCATCTTCTTCACAGTGAAGACTCACAG
GAGGAAAGCAGCCAGGACAGCAGTGGGCAGGATCGACACCCACCCCGCCACAGGGCCAACATCCT
CGAAACACCAGAAGAAGTCCAAGTTACATGGCGCCACTGAAACCTCAGGCTGTTCAGGTACCACC
CTTACTGTGGAGATGGATGAGGAGCTGCACTACGCTTCCCTCAACTTTCATGGGATGAATCCTTC
TGAGGACACCTCCACCGAATACTCAGAGGTCAGGACCCAGTGA 308. CD33 macaque aa
MPLLLLLPLLWAGALAMDPRVRLEVQESVTVQEGLCVLVPCTFFHPVPYHTRNSPVHGYWFREGA
IVSLDSPVATNKLDQEVQEETQGRFRLLGDPSRNNCSLSIVDARRRDNGSYFFRMEKGSTKYSYK
STQLSVHVTDLTHRPQILIPGALDPDHSKNLTCSVPWACEQGTPPIFSWMSAAPTSLGLRTTHSS
VLIITPRPQDHGTNLTCQVKFPGAGVTTERTIQLNVSYASQNPRTDIFLGDGSGKQGVVQGAIGG
AGVTVLLALCLCLIFFTVKTHRRKAARTAVGRIDTHPATGPTSSKHQKKSKLHGATETSGCSGTT
LTVEMDEELHYASLNFHGMNPSEDTSTEYSEVRTQ 309. 1-27 CD3-Fc + artificial
nt
ATGGGATGGAGCTGTATCATCCTCTTCTTGGTAGCAACAGCTACAGGTGTACACTCCCAAGATGG
Leader
TAATGAAGAAATGGGTGGTATTACACAGACACCATATAAAGTCTCCATCTCTGGAACCACAGTA- A
TATTGACATCCGGAGAGCCCAAATCTTGTGACAAAACTCACACATGCCCACCGTGCCCAGCACCT
GAACTCCTGGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTC
CCGGACCCCTGAGGTCACATGCGTGGTGGTGGACGTGAGCCACGAAGACCCTGAGGTCAAGTTCA
ACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAAC
AGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTA
CAAGTGCAAGGTCTCCAACAAAGCCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAG
GGCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGAGGAGATGACCAAGAACCAG
GTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAA
TGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCC
TCTATAGCAAGCTCACCGTGGACAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTG
ATGCATGAGGCTCTGCACAACCACTACACGCAGAAGAGCCTCTCCCTGTCCCCGGGTAAATAG
310. 1-27 CD3-Fc + artificial aa
MGWSCIILFLVATATGVHSQDGNEEMGGITQTPYKVSISGTTVILTSGEPKSCDKTHTCPPCPAP
Leader
ELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQY- N
STYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQ
VSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSV
MHEALHNHYTQKSLSLSPGK 311. CD33 UD H2C HL .times. artificial aa
EVQLVESGGGLVQPGGSLKLSCAASGFTFNKYAMNWVRQAPGKGLEWVARIRSKYNNYATYYADS
AF5 HL
VKDRFTISRDDSKNTAYLQMNNLKTEDTAVYYCVRHGNFGNSYISYWAYWGQGTLVTVSSGGGG- S
GGGGSGGGGSQTVVTQEPSLTVSPGGTVTLTCGSSTGAVTSGYYPNWVQQKPGQAPRGLIGGTKF
LAPGTPARFSGSLLGGKAALTLSGVQPEDEAEYYCALWYSNRWVFGGGTKLTVLSGGGGSQVQLV
QSGAEVKKPGASVKVSCKASGYTFTNYGMNWVKQAPGQGLKWMGWINTYTGEPTYADDFKGRVTM
TSDTSTSTAYLELHNLRSDDTAVYYCARWSWSDGYYVYFDYWGQGTTVTVSSGGGGSGGGGSGGG
GSDIVMTQSPDSLTVSLGERTTINCKSSQSVLDSSKNKNSLAWYQQKPGQPPKLLLSWASTRESG
IPDRFSGSGSGTDFTLTIDSLQPEDSATYYCQQSAHFPITFGQGTRLEIK 312. CD33 UD H2C
HL .times. artificial nt
GAGGTGCAGCTGGTCGAGTCTGGAGGAGGATTGGTGCAGCCTGGAGGGTCATTGAAACTCTCATG
AF5 HL
TGCAGCCTCTGGATTCACCTTCAATAAGTACGCCATGAACTGGGTCCGCCAGGCTCCAGGAAAG- G
GTTTGGAATGGGTTGCTCGCATAAGAAGTAAATATAATAATTATGCAACATATTATGCCGATTCA
GTGAAAGACAGGTTCACCATCTCCAGAGATGATTCAAAAAACACTGCCTATCTACAAATGAACAA
CTTGAAAACTGAGGACACTGCCGTGTACTACTGTGTGAGACATGGGAACTTCGGTAATAGCTACA
TATCCTACTGGGCTTACTGGGGCCAAGGGACTCTGGTCACCGTCTCCTCAGGTGGTGGTGGTTCT
GGCGGCGGCGGCTCCGGTGGTGGTGGTTCTCAGACTGTTGTGACTCAGGAACCTTCACTCACCGT
ATCACCTGGTGGAACAGTCACACTCACTTGTGGCTCCTCGACTGGGGCTGTTACATCTGGCTACT
ACCCAAACTGGGTCCAACAAAAACCAGGTCAGGCACCCCGTGGTCTAATAGGTGGGACTAAGTTC
CTCGCCCCCGGTACTCCTGCCAGATTCTCAGGCTCCCTGCTTGGAGGCAAGGCTGCCCTCACCCT
CTCAGGGGTACAGCCAGAGGATGAGGCAGAATATTACTGTGCTCTATGGTACAGCAACCGCTGGG
TGTTCGGTGGAGGAACCAAACTGACTGTCCTATCCGGAGGTGGTGGCTCCCAGGTGCAGCTGGTC
CAGTCTGGAGCTGAGGTGAAGAAGCCTGGAGCGTCAGTCAAGGTCTCCTGCAAGGCTAGCGGGTA
TACCTTCACAAACTATGGAATGAACTGGGTGAAGCAGGCTCCAGGACAGGGTTTAAAGTGGATGG
GCTGGATAAACACCTACACTGGAGAGCCAACATATGCTGATGACTTCAAGGGACGGGTTACCATG
ACTTCGGATACCTCTACCAGCACTGCCTATTTGGAACTCCACAACCTCAGAAGTGATGACACGGC
TGTATATTACTGTGCGCGCTGGAGTTGGAGTGATGGTTACTACGTTTACTTTGACTACTGGGGCC
AAGGCACTACGGTCACCGTCTCCTCAGGTGGTGGTGGTTCTGGCGGCGGCGGCTCCGGTGGTGGT
GGTTCTGACATCGTGATGACACAGTCTCCAGACTCCCTGACTGTGTCTCTGGGCGAGAGGACCAC
CATCAACTGCAAGTCCAGCCAGAGTGTTTTAGACAGCTCCAAGAATAAGAACTCCTTAGCTTGGT
ACCAGCAGAAACCAGGACAGCCTCCTAAATTACTCCTTTCCTGGGCATCTACGCGGGAATCCGGG
ATCCCTGACCGATTCAGTGGCAGCGGGTCTGGGACAGATTTCACTCTCACTATTGACAGCCTGCA
GCCTGAAGATTCTGCAACTTACTATTGTCAACAGTCTGCCCACTTCCCGATCACCTTTGGCCAAG
GGACACGACTGGAGATTAAA 313. CD33 UD F12Q HL .times. artificial aa
EVQLVESGGGLVQPGGSLKLSCAASGFTFNSYAMNWVRQAPGKGLEWVARIRSKYNNYATYYADS
AF5 HL
VKGRFTISRDDSKNTAYLQMNNLKTEDTAVYYCVRHGNFGNSYVSWWAYWGQGTLVTVSSGGGG- S
GGGGSGGGGSQTVVTQEPSLTVSPGGTVTLTCGSSTGAVTSGNYPNWVQQKPGQAPRGLIGGTKF
LAPGTPARFSGSLLGGKAALTLSGVQPEDEAEYYCVLWYSNRWVFGGGTKLTVLSGGGGSQVQLV
QSGAEVKKPGASVKVSCKASGYTFTNYGMNWVKQAPGQGLKWMGWINTYTGEPTYADDFKGRVTM
TSDTSTSTAYLELHNLRSDDTAVYYCARWSWSDGYYVYFDYWGQGTTVTVSSGGGGSGGGGSGGG
GSDIVMTQSPDSLTVSLGERTTINCKSSQSVLDSSKNKNSLAWYQQKPGQPPKLLLSWASTRESG
IPDRFSGSGSGTDFTLTIDSLQPEDSATYYCQQSAHFPITFGQGTRLEIK 314. CD33 UD
F12Q HL .times. artificial nt
GAGGTGCAGCTGGTCGAGTCTGGAGGAGGATTGGTGCAGCCTGGAGGGTCATTGAAACTCTCATG
AF5 HL
TGCAGCCTCTGGATTCACCTTCAATAGCTACGCCATGAACTGGGTCCGCCAGGCTCCAGGAAAG- G
GTTTGGAATGGGTTGCTCGCATAAGAAGTAAATATAATAATTATGCAACATATTATGCCGATTCA
GTGAAAGGCAGGTTCACCATCTCCAGAGATGATTCAAAAAACACTGCCTATCTACAAATGAACAA
CTTGAAAACTGAGGACACTGCCGTGTACTACTGTGTGAGACATGGGAACTTCGGTAATAGCTACG
TTTCCTGGTGGGCTTACTGGGGCCAAGGGACTCTGGTCACCGTCTCCTCAGGTGGTGGTGGTTCT
GGCGGCGGCGGCTCCGGTGGTGGTGGTTCTCAGACTGTTGTGACTCAGGAACCTTCACTCACCGT
ATCACCTGGTGGAACAGTCACACTCACTTGTGGCTCCTCGACTGGGGCTGTTACATCTGGCAACT
ACCCAAACTGGGTCCAACAAAAACCAGGTCAGGCACCCCGTGGTCTAATAGGTGGGACTAAGTTC
CTCGCCCCCGGTACTCCTGCCAGATTCTCAGGCTCCCTGCTTGGAGGCAAGGCTGCCCTCACCCT
CTCAGGGGTACAGCCAGAGGATGAGGCAGAATATTACTGTGTTCTATGGTACAGCAACCGCTGGG
TGTTCGGTGGAGGAACCAAACTGACTGTCCTATCCGGAGGTGGTGGCTCCCAGGTGCAGCTGGTC
CAGTCTGGAGCTGAGGTGAAGAAGCCTGGAGCGTCAGTCAAGGTCTCCTGCAAGGCTAGCGGGTA
TACCTTCACAAACTATGGAATGAACTGGGTGAAGCAGGCTCCAGGACAGGGTTTAAAGTGGATGG
GCTGGATAAACACCTACACTGGAGAGCCAACATATGCTGATGACTTCAAGGGACGGGTTACCATG
ACTTCGGATACCTCTACCAGCACTGCCTATTTGGAACTCCACAACCTCAGAAGTGATGACACGGC
TGTATATTACTGTGCGCGCTGGAGTTGGAGTGATGGTTACTACGTTTACTTTGACTACTGGGGCC
AAGGCACTACGGTCACCGTCTCCTCAGGTGGTGGTGGTTCTGGCGGCGGCGGCTCCGGTGGTGGT
GGTTCTGACATCGTGATGACACAGTCTCCAGACTCCCTGACTGTGTCTCTGGGCGAGAGGACCAC
CATCAACTGCAAGTCCAGCCAGAGTGTTTTAGACAGCTCCAAGAATAAGAACTCCTTAGCTTGGT
ACCAGCAGAAACCAGGACAGCCTCCTAAATTACTCCTTTCCTGGGCATCTACGCGGGAATCCGGG
ATCCCTGACCGATTCAGTGGCAGCGGGTCTGGGACAGATTTCACTCTCACTATTGACAGCCTGCA
GCCTGAAGATTCTGCAACTTACTATTGTCAACAGTCTGCCCACTTCCCGATCACCTTTGGCCAAG
GGACACGACTGGAGATTAAA 315. CD33 UD I2C HL .times. artificial aa
EVQLVESGGGLVQPGGSLKLSCAASGFTFNKYAMNWVRQAPGKGLEWVARIRSKYNNYATYYADS
AF5 HL
VKDRFTISRDDSKNTAYLQMNNLKTEDTAVYYCVRHGNFGNSYISYWAYWGQGTLVTVSSGGGG- S
GGGGSGGGGSQTVVTQEPSLTVSPGGTVTLTCGSSTGAVTSGNYPNWVQQKPGQAPRGLIGGTKF
LAPGTPARFSGSLLGGKAALTLSGVQPEDEAEYYCVLWYSNRWVFGGGTKLTVLSGGGGSQVQLV
QSGAEVKKPGASVKVSCKASGYTFTNYGMNWVKQAPGQGLKWMGWINTYTGEPTYADDFKGRVTM
TSDTSTSTAYLELHNLRSDDTAVYYCARWSWSDGYYVYFDYWGQGTTVTVSSGGGGSGGGGSGGG
GSDIVMTQSPDSLTVSLGERTTINCKSSQSVLDSSKNKNSLAWYQQKPGQPPKLLLSWASTRESG
IPDRFSGSGSGTDFTLTIDSLQPEDSATYYCQQSAHFPITFGQGTRLEIK 316. CD33 UD I2C
HL .times. artificial nt
GAGGTGCAGCTGGTCGAGTCTGGAGGAGGATTGGTGCAGCCTGGAGGGTCATTGAAACTCTCATG
AF5 HL
TGCAGCCTCTGGATTCACCTTCAATAAGTACGCCATGAACTGGGTCCGCCAGGCTCCAGGAAAG- G
GTTTGGAATGGGTTGCTCGCATAAGAAGTAAATATAATAATTATGCAACATATTATGCCGATTCA
GTGAAAGACAGGTTCACCATCTCCAGAGATGATTCAAAAAACACTGCCTATCTACAAATGAACAA
CTTGAAAACTGAGGACACTGCCGTGTACTACTGTGTGAGACATGGGAACTTCGGTAATAGCTACA
TATCCTACTGGGCTTACTGGGGCCAAGGGACTCTGGTCACCGTCTCCTCAGGTGGTGGTGGTTCT
GGCGGCGGCGGCTCCGGTGGTGGTGGTTCTCAGACTGTTGTGACTCAGGAACCTTCACTCACCGT
ATCACCTGGTGGAACAGTCACACTCACTTGTGGCTCCTCGACTGGGGCTGTTACATCTGGCAACT
ACCCAAACTGGGTCCAACAAAAACCAGGTCAGGCACCCCGTGGTCTAATAGGTGGGACTAAGTTC
CTCGCCCCCGGTACTCCTGCCAGATTCTCAGGCTCCCTGCTTGGAGGCAAGGCTGCCCTCACCCT
CTCAGGGGTACAGCCAGAGGATGAGGCAGAATATTACTGTGTTCTATGGTACAGCAACCGCTGGG
TGTTCGGTGGAGGAACCAAACTGACTGTCCTATCCGGAGGTGGTGGCTCCCAGGTGCAGCTGGTC
CAGTCTGGAGCTGAGGTGAAGAAGCCTGGAGCGTCAGTCAAGGTCTCCTGCAAGGCTAGCGGGTA
TACCTTCACAAACTATGGAATGAACTGGGTGAAGCAGGCTCCAGGACAGGGTTTAAAGTGGATGG
GCTGGATAAACACCTACACTGGAGAGCCAACATATGCTGATGACTTCAAGGGACGGGTTACCATG
ACTTCGGATACCTCTACCAGCACTGCCTATTTGGAACTCCACAACCTCAGAAGTGATGACACGGC
TGTATATTACTGTGCGCGCTGGAGTTGGAGTGATGGTTACTACGTTTACTTTGACTACTGGGGCC
AAGGCACTACGGTCACCGTCTCCTCAGGTGGTGGTGGTTCTGGCGGCGGCGGCTCCGGTGGTGGT
GGTTCTGACATCGTGATGACACAGTCTCCAGACTCCCTGACTGTGTCTCTGGGCGAGAGGACCAC
CATCAACTGCAAGTCCAGCCAGAGTGTTTTAGACAGCTCCAAGAATAAGAACTCCTTAGCTTGGT
ACCAGCAGAAACCAGGACAGCCTCCTAAATTACTCCTTTCCTGGGCATCTACGCGGGAATCCGGG
ATCCCTGACCGATTCAGTGGCAGCGGGTCTGGGACAGATTTCACTCTCACTATTGACAGCCTGCA
GCCTGAAGATTCTGCAACTTACTATTGTCAACAGTCTGCCCACTTCCCGATCACCTTTGGCCAAG
GGACACGACTGGAGATTAAA 317. MCSP-A9 HL .times. H2C artificial aa
QVQLVQSGAEVKKPGASVKVSCKASGYPFTGYYMHWVRQAPGQGLEWMGWINPNSGGTNYAQKFQ
HL
GRVTITADESTSTAYMELSRLRSDDTAVYYCAKSWVSWFASWGQGTLVTVSSGGGGSGGGGSGGG
GSDIVMTQSPDSLAVSLGERATINCKSSQSVLSSSNNKNYLNWYQQKPGQPPKLLIYWASTRESG
VPDRFSGSGSGTDFTLTISSLQAEDVAVYYCQQHFNTPFAFGQGTKLEIKSGGGGSEVQLVESGG
GLVQPGGSLKLSCAASGFTFNKYAMNWVRQAPGKGLEWVARIRSKYNNYATYYADSVKDRFTISR
DDSKNTAYLQMNNLKTEDTAVYYCVRHGNFGNSYISYWAYWGQGTLVTVSSGGGGSGGGGSGGGG
SQTVVTQEPSLTVSPGGTVTLTCGSSTGAVTSGYYPNWVQQKPGQAPRGLIGGTKFLAPGTPARF
SGSLLGGKAALTLSGVQPEDEAEYYCALWYSNRWVFGGGTKLTVL 318. MCSP-A9 HL
.times. H2C artificial nt
CAGGTGCAGCTGGTCCAGTCTGGGGCTGAGGTGAAGAAGCCTGGGGCCTCAGTGAAGGTCTCCTG
HL
CAAGGCTTCTGGATACCCCTTCACCGGCTACTACATGCACTGGGTGCGACAGGCCCCTGGACAAG
GGCTTGAGTGGATGGGATGGATCAACCCTAACAGTGGTGGCACAAACTATGCACAGAAGTTTCAG
GGCAGAGTCACGATTACCGCGGACGAATCCACGAGCACAGCCTACATGGAGCTGAGCAGGCTGAG
ATCTGACGACACGGCCGTGTATTACTGTGCGAAATCCTGGGTCTCCTGGTTTGCTTCCTGGGGTC
AAGGAACCTTGGTCACCGTCTCCTCAGGTGGTGGTGGTTCTGGCGGCGGCGGCTCCGGTGGTGGT
GGTTCTGACATCGTGATGACACAGTCTCCAGACTCCCTGGCTGTGTCTCTGGGCGAGAGGGCCAC
CATCAACTGCAAGTCCAGCCAGAGTGTCTTATCCAGCTCCAACAATAAGAACTACTTAAATTGGT
ACCAGCAGAAACCAGGACAGCCTCCTAAGTTGCTCATTTACTGGGCATCTACCCGGGAATCCGGG
GTCCCTGACCGATTCAGTGGCAGCGGGTCTGGGACAGATTTCACTCTCACCATCAGCAGCCTGCA
GGCTGAAGATGTGGCGGTTTATTACTGTCAACAACATTTTAATACTCCGTTCGCTTTTGGCCAGG
GGACCAAGCTGGAGATCAAATCCGGAGGTGGTGGATCCGAGGTGCAGCTGGTCGAGTCTGGAGGA
GGATTGGTGCAGCCTGGAGGGTCATTGAAACTCTCATGTGCAGCCTCTGGATTCACCTTCAATAA
GTACGCCATGAACTGGGTCCGCCAGGCTCCAGGAAAGGGTTTGGAATGGGTTGCTCGCATAAGAA
GTAAATATAATAATTATGCAACATATTATGCCGATTCAGTGAAAGACAGGTTCACCATCTCCAGA
GATGATTCAAAAAACACTGCCTATCTACAAATGAACAACTTGAAAACTGAGGACACTGCCGTGTA
CTACTGTGTGAGACATGGGAACTTCGGTAATAGCTACATATCCTACTGGGCTTACTGGGGCCAAG
GGACTCTGGTCACCGTCTCCTCAGGTGGTGGTGGTTCTGGCGGCGGCGGCTCCGGTGGTGGTGGT
TCTCAGACTGTTGTGACTCAGGAACCTTCACTCACCGTATCACCTGGTGGAACAGTCACACTCAC
TTGTGGCTCCTCGACTGGGGCTGTTACATCTGGCTACTACCCAAACTGGGTCCAACAAAAACCAG
GTCAGGCACCCCGTGGTCTAATAGGTGGGACTAAGTTCCTCGCCCCCGGTACTCCTGCCAGATTC
TCAGGCTCCCTGCTTGGAGGCAAGGCTGCCCTCACCCTCTCAGGGGTACAGCCAGAGGATGAGGC
AGAATATTACTGTGCTCTATGGTACAGCAACCGCTGGGTGTTCGGTGGAGGAACCAAACTGACTG
TCCTA 319. MCSP-A9 HL .times. artificial aa
QVQLVQSGAEVKKPGASVKVSCKASGYPFTGYYMHWVRQAPGQGLEWMGWINPNSGGTNYAQKFQ
F12Q HL
GRVTITADESTSTAYMELSRLRSDDTAVYYCAKSWVSWFASWGQGTLVTVSSGGGGSGGGGSG- GG
GSDIVMTQSPDSLAVSLGERATINCKSSQSVLSSSNNKNYLNWYQQKPGQPPKLLIYWASTRESG
VPDRFSGSGSGTDFTLTISSLQAEDVAVYYCQQHFNTPFAFGQGTKLEIKSGGGGSEVQLVESGG
GLVQPGGSLKLSCAASGFTFNSYAMNWVRQAPGKGLEWVARIRSKYNNYATYYADSVKGRFTISR
DDSKNTAYLQMNNLKTEDTAVYYCVRHGNFGNSYVSWWAYWGQGTLVTVSSGGGGSGGGGSGGGG
SQTVVTQEPSLTVSPGGTVTLTCGSSTGAVTSGNYPNWVQQKPGQAPRGLIGGTKFLAPGTPARF
SGSLLGGKAALTLSGVQPEDEAEYYCVLWYSNRWVFGGGTKLTVL 320. MCSP-A9 HL
.times. artificial nt
CAGGTGCAGCTGGTCCAGTCTGGGGCTGAGGTGAAGAAGCCTGGGGCCTCAGTGAAGGTCTCCTG
F12Q HL
CAAGGCTTCTGGATACCCCTTCACCGGCTACTACATGCACTGGGTGCGACAGGCCCCTGGACA- AG
GGCTTGAGTGGATGGGATGGATCAACCCTAACAGTGGTGGCACAAACTATGCACAGAAGTTTCAG
GGCAGAGTCACGATTACCGCGGACGAATCCACGAGCACAGCCTACATGGAGCTGAGCAGGCTGAG
ATCTGACGACACGGCCGTGTATTACTGTGCGAAATCCTGGGTCTCCTGGTTTGCTTCCTGGGGTC
AAGGAACCTTGGTCACCGTCTCCTCAGGTGGTGGTGGTTCTGGCGGCGGCGGCTCCGGTGGTGGT
GGTTCTGACATCGTGATGACACAGTCTCCAGACTCCCTGGCTGTGTCTCTGGGCGAGAGGGCCAC
CATCAACTGCAAGTCCAGCCAGAGTGTCTTATCCAGCTCCAACAATAAGAACTACTTAAATTGGT
ACCAGCAGAAACCAGGACAGCCTCCTAAGTTGCTCATTTACTGGGCATCTACCCGGGAATCCGGG
GTCCCTGACCGATTCAGTGGCAGCGGGTCTGGGACAGATTTCACTCTCACCATCAGCAGCCTGCA
GGCTGAAGATGTGGCGGTTTATTACTGTCAACAACATTTTAATACTCCGTTCGCTTTTGGCCAGG
GGACCAAGCTGGAGATCAAATCCGGAGGTGGTGGATCCGAGGTGCAGCTGGTCGAGTCTGGAGGA
GGATTGGTGCAGCCTGGAGGGTCATTGAAACTCTCATGTGCAGCCTCTGGATTCACCTTCAATAG
CTACGCCATGAACTGGGTCCGCCAGGCTCCAGGAAAGGGTTTGGAATGGGTTGCTCGCATAAGAA
GTAAATATAATAATTATGCAACATATTATGCCGATTCAGTGAAAGGCAGGTTCACCATCTCCAGA
GATGATTCAAAAAACACTGCCTATCTACAAATGAACAACTTGAAAACTGAGGACACTGCCGTGTA
CTACTGTGTGAGACATGGGAACTTCGGTAATAGCTACGTTTCCTGGTGGGCTTACTGGGGCCAAG
GGACTCTGGTCACCGTCTCCTCAGGTGGTGGTGGTTCTGGCGGCGGCGGCTCCGGTGGTGGTGGT
TCTCAGACTGTTGTGACTCAGGAACCTTCACTCACCGTATCACCTGGTGGAACAGTCACACTCAC
TTGTGGCTCCTCGACTGGGGCTGTTACATCTGGCAACTACCCAAACTGGGTCCAACAAAAACCAG
GTCAGGCACCCCGTGGTCTAATAGGTGGGACTAAGTTCCTCGCCCCCGGTACTCCTGCCAGATTC
TCAGGCTCCCTGCTTGGAGGCAAGGCTGCCCTCACCCTCTCAGGGGTACAGCCAGAGGATGAGGC
AGAATATTACTGTGTTCTATGGTACAGCAACCGCTGGGTGTTCGGTGGAGGAACCAAACTGACTG
TCCTA 321. MCSP-A9 HL .times. I2C artificial aa
QVQLVQSGAEVKKPGASVKVSCKASGYPFTGYYMHWVRQAPGQGLEWMGWINPNSGGTNYAQKFQ
HL
GRVTITADESTSTAYMELSRLRSDDTAVYYCAKSWVSWFASWGQGTLVTVSSGGGGSGGGGSGGG
GSDIVMTQSPDSLAVSLGERATINCKSSQSVLSSSNNKNYLNWYQQKPGQPPKLLIYWASTRESG
VPDRFSGSGSGTDFTLTISSLQAEDVAVYYCQQHFNTPFAFGQGTKLEIKSGGGGSEVQLVESGG
GLVQPGGSLKLSCAASGFTFNKYAMNWVRQAPGKGLEWVARIRSKYNNYATYYADSVKDRFTISR
DDSKNTAYLQMNNLKTEDTAVYYCVRHGNFGNSYISYWAYWGQGTLVTVSSGGGGSGGGGSGGGG
SQTVVTQEPSLTVSPGGTVTLTCGSSTGAVTSGNYPNWVQQKPGQAPRGLIGGTKFLAPGTPARF
SGSLLGGKAALTLSGVQPEDEAEYYCVLWYSNRWVFGGGTKLTVL 322. MCSP-A9 HL
.times. I2C artificial nt
CAGGTGCAGCTGGTCCAGTCTGGGGCTGAGGTGAAGAAGCCTGGGGCCTCAGTGAAGGTCTCCTG
HL
CAAGGCTTCTGGATACCCCTTCACCGGCTACTACATGCACTGGGTGCGACAGGCCCCTGGACAAG
GGCTTGAGTGGATGGGATGGATCAACCCTAACAGTGGTGGCACAAACTATGCACAGAAGTTTCAG
GGCAGAGTCACGATTACCGCGGACGAATCCACGAGCACAGCCTACATGGAGCTGAGCAGGCTGAG
ATCTGACGACACGGCCGTGTATTACTGTGCGAAATCCTGGGTCTCCTGGTTTGCTTCCTGGGGTC
AAGGAACCTTGGTCACCGTCTCCTCAGGTGGTGGTGGTTCTGGCGGCGGCGGCTCCGGTGGTGGT
GGTTCTGACATCGTGATGACACAGTCTCCAGACTCCCTGGCTGTGTCTCTGGGCGAGAGGGCCAC
CATCAACTGCAAGTCCAGCCAGAGTGTCTTATCCAGCTCCAACAATAAGAACTACTTAAATTGGT
ACCAGCAGAAACCAGGACAGCCTCCTAAGTTGCTCATTTACTGGGCATCTACCCGGGAATCCGGG
GTCCCTGACCGATTCAGTGGCAGCGGGTCTGGGACAGATTTCACTCTCACCATCAGCAGCCTGCA
GGCTGAAGATGTGGCGGTTTATTACTGTCAACAACATTTTAATACTCCGTTCGCTTTTGGCCAGG
GGACCAAGCTGGAGATCAAATCCGGAGGTGGTGGATCCGAGGTGCAGCTGGTCGAGTCTGGAGGA
GGATTGGTGCAGCCTGGAGGGTCATTGAAACTCTCATGTGCAGCCTCTGGATTCACCTTCAATAA
GTACGCCATGAACTGGGTCCGCCAGGCTCCAGGAAAGGGTTTGGAATGGGTTGCTCGCATAAGAA
GTAAATATAATAATTATGCAACATATTATGCCGATTCAGTGAAAGACAGGTTCACCATCTCCAGA
GATGATTCAAAAAACACTGCCTATCTACAAATGAACAACTTGAAAACTGAGGACACTGCCGTGTA
CTACTGTGTGAGACATGGGAACTTCGGTAATAGCTACATATCCTACTGGGCTTACTGGGGCCAAG
GGACTCTGGTCACCGTCTCCTCAGGTGGTGGTGGTTCTGGCGGCGGCGGCTCCGGTGGTGGTGGT
TCTCAGACTGTTGTGACTCAGGAACCTTCACTCACCGTATCACCTGGTGGAACAGTCACACTCAC
TTGTGGCTCCTCGACTGGGGCTGTTACATCTGGCAACTACCCAAACTGGGTCCAACAAAAACCAG
GTCAGGCACCCCGTGGTCTAATAGGTGGGACTAAGTTCCTCGCCCCCGGTACTCCTGCCAGATTC
TCAGGCTCCCTGCTTGGAGGCAAGGCTGCCCTCACCCTCTCAGGGGTACAGCCAGAGGATGAGGC
AGAATATTACTGTGTTCTATGGTACAGCAACCGCTGGGTGTTCGGTGGAGGAACCAAACTGACTG
TCCTA 323. MCSP-C8 HL .times. I2C artificial aa
QVQLVQSGAEVKRPGASMKVSCKASGYTFTNYYIHWVRQAPGQGLEWMGWINPNSGATNYAQKFQ
HL
GRVTMTRDTSTSTVYMELSSLRSEDTAVYYCAKSWVSWFASWGQGTLVTVSSGGGGSGGGGSGGG
GSDIVMTQSPDSLAVSLGERATINCKSSQSVLNSKNNRNYLAWYQQKPGQPPKLLIYWASTRESG
VPDRFSGSGSGTDFTLTISGLQAEDVAVYYCQQHYSTPFTFGPGTKVDIKSGGGGSEVQLVESGG
GLVQPGGSLKLSCAASGFTFNKYAMNWVRQAPGKGLEWVARIRSKYNNYATYYADSVKDRFTISR
DDSKNTAYLQMNNLKTEDTAVYYCVRHGNFGNSYISYWAYWGQGTLVTVSSGGGGSGGGGSGGGG
SQTVVTQEPSLTVSPGGTVTLTCGSSTGAVTSGNYPNWVQQKPGQAPRGLIGGTKFLAPGTPARF
SGSLLGGKAALTLSGVQPEDEAEYYCVLWYSNRWVFGGGTKLTVL 324. MCSP-C8 HL
.times. I2C artificial nt
CAGGTGCAGCTGGTCCAGTCTGGGGCTGAGGTGAAGAGGCCTGGGGCCTCAATGAAGGTCTCCTG
HL
CAAGGCTTCTGGGTACACCTTCACCAACTACTATATACACTGGGTGCGACAGGCCCCTGGACAAG
GTCTTGAGTGGATGGGTTGGATCAACCCTAACAGTGGTGCCACAAACTATGCACAGAAGTTCCAG
GGCAGAGTCACCATGACCAGGGACACGTCCACGAGCACAGTCTACATGGAGCTGAGCAGCCTGAG
ATCTGAGGACACGGCCGTGTATTACTGTGCGAAATCCTGGGTCTCCTGGTTTGCTTCCTGGGGTC
AAGGAACCTTGGTCACCGTCTCCTCAGGTGGTGGTGGTTCTGGCGGCGGCGGCTCCGGTGGTGGT
GGTTCTGACATCGTGATGACCCAGTCTCCAGACTCCCTGGCTGTGTCTCTGGGCGAGAGGGCCAC
CATCAACTGCAAGTCCAGCCAGAGTGTTTTAAACAGCAAGAACAATAGGAACTACTTAGCTTGGT
ACCAGCAGAAACCAGGACAGCCTCCTAAGCTGCTCATTTACTGGGCATCTACCCGGGAATCCGGG
GTCCCTGACCGATTCAGTGGCAGCGGGTCTGGGACAGATTTCACTCTCACCATCAGTGGCCTGCA
GGCTGAAGATGTGGCAGTTTATTACTGTCAGCAACATTATAGTACTCCATTCACTTTTGGCCCTG
GGACCAAAGTGGATATCAAATCCGGAGGTGGTGGATCCGAGGTGCAGCTGGTCGAGTCTGGAGGA
GGATTGGTGCAGCCTGGAGGGTCATTGAAACTCTCATGTGCAGCCTCTGGATTCACCTTCAATAA
GTACGCCATGAACTGGGTCCGCCAGGCTCCAGGAAAGGGTTTGGAATGGGTTGCTCGCATAAGAA
GTAAATATAATAATTATGCAACATATTATGCCGATTCAGTGAAAGACAGGTTCACCATCTCCAGA
GATGATTCAAAAAACACTGCCTATCTACAAATGAACAACTTGAAAACTGAGGACACTGCCGTGTA
CTACTGTGTGAGACATGGGAACTTCGGTAATAGCTACATATCCTACTGGGCTTACTGGGGCCAAG
GGACTCTGGTCACCGTCTCCTCAGGTGGTGGTGGTTCTGGCGGCGGCGGCTCCGGTGGTGGTGGT
TCTCAGACTGTTGTGACTCAGGAACCTTCACTCACCGTATCACCTGGTGGAACAGTCACACTCAC
TTGTGGCTCCTCGACTGGGGCTGTTACATCTGGCAACTACCCAAACTGGGTCCAACAAAAACCAG
GTCAGGCACCCCGTGGTCTAATAGGTGGGACTAAGTTCCTCGCCCCCGGTACTCCTGCCAGATTC
TCAGGCTCCCTGCTTGGAGGCAAGGCTGCCCTCACCCTCTCAGGGGTACAGCCAGAGGATGAGGC
AGAATATTACTGTGTTCTATGGTACAGCAACCGCTGGGTGTTCGGTGGAGGAACCAAACTGACTG
TCCTA 325. MCSP-B8 HL .times. I2C artificial aa
QVQLVQSGAEVKRPGASMKVSCKASGYTFTNYYIHWVRQAPGQGLEWMGWINPNSGATNYAQKFQ
HL
GRVTMTRDTSTSTVYMELSSLRSEDTAVYYCAKSWVSWFASWGQGTLVTVSSGGGGSGGGGSGGG
GSDIVMTQSPDSLTVSPGERATINCKSSQSVLNSKNNRNYLAWYQQKPGQPPKLLIYWASTRESG
VPDRFSGSGSGTDFTLTIDSLQPEDSATYYCQQHYSTPFTFGQGTRLEIKSGGGGSEVQLVESGG
GLVQPGGSLKLSCAASGFTFNKYAMNWVRQAPGKGLEWVARIRSKYNNYATYYADSVKDRFTISR
DDSKNTAYLQMNNLKTEDTAVYYCVRHGNFGNSYISYWAYWGQGTLVTVSSGGGGSGGGGSGGGG
SQTVVTQEPSLTVSPGGTVTLTCGSSTGAVTSGNYPNWVQQKPGQAPRGLIGGTKFLAPGTPARF
SGSLLGGKAALTLSGVQPEDEAEYYCVLWYSNRWVFGGGTKLTVL 326. MCSP-B8 HL
.times. I2C artificial nt
CAGGTGCAGCTGGTCCAGTCTGGGGCTGAGGTGAAGAGGCCTGGGGCCTCAATGAAGGTCTCCTG
HL
CAAGGCTTCTGGGTACACCTTCACCAACTACTATATACACTGGGTGCGACAGGCCCCTGGACAAG
GTCTTGAGTGGATGGGTTGGATCAACCCTAACAGTGGTGCCACAAACTATGCACAGAAGTTCCAG
GGCAGAGTCACCATGACCAGGGACACGTCCACGAGCACAGTCTACATGGAGCTGAGCAGCCTGAG
ATCTGAGGACACGGCCGTGTATTACTGTGCGAAATCCTGGGTCTCCTGGTTTGCTTCCTGGGGTC
AAGGAACCTTGGTCACCGTCTCCTCAGGTGGTGGTGGTTCTGGCGGCGGCGGCTCCGGTGGTGGT
GGTTCTGACATCGTGATGACCCAGTCTCCAGACTCCCTGACTGTGTCTCCGGGCGAGAGGGCCAC
CATCAACTGCAAGTCCAGCCAGAGTGTTTTAAACAGCAAGAACAATAGGAACTACTTAGCTTGGT
ACCAGCAGAAACCAGGACAGCCTCCTAAGCTGCTCATTTACTGGGCATCTACCCGGGAATCCGGG
GTCCCTGACCGATTCAGTGGCAGCGGGTCTGGGACAGATTTCACTCTCACCATCGATTCCCTGCA
GCCTGAAGATAGTGCAACTTATTACTGTCAGCAACATTATAGTACTCCATTCACTTTTGGCCAGG
GGACCAGACTGGAGATCAAATCCGGAGGTGGTGGATCCGAGGTGCAGCTGGTCGAGTCTGGAGGA
GGATTGGTGCAGCCTGGAGGGTCATTGAAACTCTCATGTGCAGCCTCTGGATTCACCTTCAATAA
GTACGCCATGAACTGGGTCCGCCAGGCTCCAGGAAAGGGTTTGGAATGGGTTGCTCGCATAAGAA
GTAAATATAATAATTATGCAACATATTATGCCGATTCAGTGAAAGACAGGTTCACCATCTCCAGA
GATGATTCAAAAAACACTGCCTATCTACAAATGAACAACTTGAAAACTGAGGACACTGCCGTGTA
CTACTGTGTGAGACATGGGAACTTCGGTAATAGCTACATATCCTACTGGGCTTACTGGGGCCAAG
GGACTCTGGTCACCGTCTCCTCAGGTGGTGGTGGTTCTGGCGGCGGCGGCTCCGGTGGTGGTGGT
TCTCAGACTGTTGTGACTCAGGAACCTTCACTCACCGTATCACCTGGTGGAACAGTCACACTCAC
TTGTGGCTCCTCGACTGGGGCTGTTACATCTGGCAACTACCCAAACTGGGTCCAACAAAAACCAG
GTCAGGCACCCCGTGGTCTAATAGGTGGGACTAAGTTCCTCGCCCCCGGTACTCCTGCCAGATTC
TCAGGCTCCCTGCTTGGAGGCAAGGCTGCCCTCACCCTCTCAGGGGTACAGCCAGAGGATGAGGC
AGAATATTACTGTGTTCTATGGTACAGCAACCGCTGGGTGTTCGGTGGAGGAACCAAACTGACTG
TCCTA 327. MCSP-B7 HL .times. I2C artificial aa
QVQLVQSGAEVKRPGASMKVSCKASGYTFTNYYIHWVRQAPGQGLEWMGWINPNSGATNYAQKFQ
HL
GRVTMTRDTSTSTVYMELSSLRSEDTAVYYCAKSWVSWFASWGQGTLVTVSSGGGGSGGGGSGGG
GSDIVMTQSPDSLTVSLGERTTINCKSSQSVLNSKNNRNYLAWYQQKPGQPPKLLIYWASTRESG
VPDRFSGSGSGTDFTLTIDSLQPEDIATYYCQQHYSTPFTFGQGTRLEIKSGGGGSEVQLVESGG
GLVQPGGSLKLSCAASGFTFNKYAMNWVRQAPGKGLEWVARIRSKYNNYATYYADSVKDRFTISR
DDSKNTAYLQMNNLKTEDTAVYYCVRHGNFGNSYISYWAYWGQGTLVTVSSGGGGSGGGGSGGGG
SQTVVTQEPSLTVSPGGTVTLTCGSSTGAVTSGNYPNWVQQKPGQAPRGLIGGTKFLAPGTPARF
SGSLLGGKAALTLSGVQPEDEAEYYCVLWYSNRWVFGGGTKLTVL 328. MCSP-B7 HL
.times. I2C artificial nt
CAGGTGCAGCTGGTCCAGTCTGGGGCTGAGGTGAAGAGGCCTGGGGCCTCAATGAAGGTCTCCTG
HL
CAAGGCTTCTGGGTACACCTTCACCAACTACTATATACACTGGGTGCGACAGGCCCCTGGACAAG
GTCTTGAGTGGATGGGTTGGATCAACCCTAACAGTGGTGCCACAAACTATGCACAGAAGTTCCAG
GGCAGAGTCACCATGACCAGGGACACGTCCACGAGCACAGTCTACATGGAGCTGAGCAGCCTGAG
ATCTGAGGACACGGCCGTGTATTACTGTGCGAAATCCTGGGTCTCCTGGTTTGCTTCCTGGGGTC
AAGGAACCTTGGTCACCGTCTCCTCAGGTGGTGGTGGTTCTGGCGGCGGCGGCTCCGGTGGTGGT
GGTTCTGACATCGTGATGACCCAGTCTCCAGACTCCCTGACTGTGTCTCTGGGCGAGAGGACCAC
CATCAACTGCAAGTCCAGCCAGAGTGTTTTAAACAGCAAGAACAATAGGAACTACTTAGCTTGGT
ACCAGCAGAAACCAGGACAGCCTCCTAAGCTGCTCATTTACTGGGCATCTACCCGGGAATCCGGG
GTCCCTGACCGATTCAGTGGCAGCGGGTCTGGGACAGATTTCACTCTCACCATCGATTCCCTGCA
GCCTGAAGATATTGCAACTTATTACTGTCAGCAACATTATAGTACTCCATTCACTTTTGGCCAGG
GGACCAGACTGGAGATCAAATCCGGAGGTGGTGGATCCGAGGTGCAGCTGGTCGAGTCTGGAGGA
GGATTGGTGCAGCCTGGAGGGTCATTGAAACTCTCATGTGCAGCCTCTGGATTCACCTTCAATAA
GTACGCCATGAACTGGGTCCGCCAGGCTCCAGGAAAGGGTTTGGAATGGGTTGCTCGCATAAGAA
GTAAATATAATAATTATGCAACATATTATGCCGATTCAGTGAAAGACAGGTTCACCATCTCCAGA
GATGATTCAAAAAACACTGCCTATCTACAAATGAACAACTTGAAAACTGAGGACACTGCCGTGTA
CTACTGTGTGAGACATGGGAACTTCGGTAATAGCTACATATCCTACTGGGCTTACTGGGGCCAAG
GGACTCTGGTCACCGTCTCCTCAGGTGGTGGTGGTTCTGGCGGCGGCGGCTCCGGTGGTGGTGGT
TCTCAGACTGTTGTGACTCAGGAACCTTCACTCACCGTATCACCTGGTGGAACAGTCACACTCAC
TTGTGGCTCCTCGACTGGGGCTGTTACATCTGGCAACTACCCAAACTGGGTCCAACAAAAACCAG
GTCAGGCACCCCGTGGTCTAATAGGTGGGACTAAGTTCCTCGCCCCCGGTACTCCTGCCAGATTC
TCAGGCTCCCTGCTTGGAGGCAAGGCTGCCCTCACCCTCTCAGGGGTACAGCCAGAGGATGAGGC
AGAATATTACTGTGTTCTATGGTACAGCAACCGCTGGGTGTTCGGTGGAGGAACCAAACTGACTG
TCCTA 329. MCSP-G8 HL .times. I2C artificial aa
QVQLVQSGAEVKRPGASMKVSCKASGYTFTNYYIHWVRQAPGQGLEWMGWINPNSGATNYAQKFQ
HL
GRVTMTRDTSTSTVYMELSSLRSEDTAVYYCAKSWVSWFASWGQGTLVTVSSGGGGSGGGGSGGG
GSDIVMTQSPDSLTVSLGERATINCKSSQSVLNSKNNRNYLAWYQQKPGQPPKLLIYWASTRESG
VPDRFSGSGSGTDFTLTIDSLQPEDSATYYCQQHYSTPFTFGQGTRLEIKSGGGGSEVQLVESGG
GLVQPGGSLKLSCAASGFTFNKYAMNWVRQAPGKGLEWVARIRSKYNNYATYYADSVKDRFTISR
DDSKNTAYLQMNNLKTEDTAVYYCVRHGNFGNSYISYWAYWGQGTLVTVSSGGGGSGGGGSGGGG
SQTVVTQEPSLTVSPGGTVTLTCGSSTGAVTSGNYPNWVQQKPGQAPRGLIGGTKFLAPGTPARF
SGSLLGGKAALTLSGVQPEDEAEYYCVLWYSNRWVFGGGTKLTVL 330. MCSP-G8 HL
.times. I2C artificial nt
CAGGTGCAGCTGGTCCAGTCTGGGGCTGAGGTGAAGAGGCCTGGGGCCTCAATGAAGGTCTCCTG
HL
CAAGGCTTCTGGGTACACCTTCACCAACTACTATATACACTGGGTGCGACAGGCCCCTGGACAAG
GTCTTGAGTGGATGGGTTGGATCAACCCTAACAGTGGTGCCACAAACTATGCACAGAAGTTCCAG
GGCAGAGTCACCATGACCAGGGACACGTCCACGAGCACAGTCTACATGGAGCTGAGCAGCCTGAG
ATCTGAGGACACGGCCGTGTATTACTGTGCGAAATCCTGGGTCTCCTGGTTTGCTTCCTGGGGTC
AAGGAACCTTGGTCACCGTCTCCTCAGGTGGTGGTGGTTCTGGCGGCGGCGGCTCCGGTGGTGGT
GGTTCTGACATCGTGATGACCCAGTCTCCAGACTCCCTGACTGTGTCTCTGGGCGAGAGGGCCAC
CATCAACTGCAAGTCCAGCCAGAGTGTTTTAAACAGCAAGAACAATAGGAACTACTTAGCTTGGT
ACCAGCAGAAACCAGGACAGCCTCCTAAGCTGCTCATTTACTGGGCATCTACCCGGGAATCCGGG
GTCCCTGACCGATTCAGTGGCAGCGGGTCTGGGACAGATTTCACTCTCACCATCGATTCCCTGCA
GCCTGAAGATAGTGCAACTTATTACTGTCAGCAACATTATAGTACTCCATTCACTTTTGGCCAGG
GGACCAGACTGGAGATCAAATCCGGAGGTGGTGGATCCGAGGTGCAGCTGGTCGAGTCTGGAGGA
GGATTGGTGCAGCCTGGAGGGTCATTGAAACTCTCATGTGCAGCCTCTGGATTCACCTTCAATAA
GTACGCCATGAACTGGGTCCGCCAGGCTCCAGGAAAGGGTTTGGAATGGGTTGCTCGCATAAGAA
GTAAATATAATAATTATGCAACATATTATGCCGATTCAGTGAAAGACAGGTTCACCATCTCCAGA
GATGATTCAAAAAACACTGCCTATCTACAAATGAACAACTTGAAAACTGAGGACACTGCCGTGTA
CTACTGTGTGAGACATGGGAACTTCGGTAATAGCTACATATCCTACTGGGCTTACTGGGGCCAAG
GGACTCTGGTCACCGTCTCCTCAGGTGGTGGTGGTTCTGGCGGCGGCGGCTCCGGTGGTGGTGGT
TCTCAGACTGTTGTGACTCAGGAACCTTCACTCACCGTATCACCTGGTGGAACAGTCACACTCAC
TTGTGGCTCCTCGACTGGGGCTGTTACATCTGGCAACTACCCAAACTGGGTCCAACAAAAACCAG
GTCAGGCACCCCGTGGTCTAATAGGTGGGACTAAGTTCCTCGCCCCCGGTACTCCTGCCAGATTC
TCAGGCTCCCTGCTTGGAGGCAAGGCTGCCCTCACCCTCTCAGGGGTACAGCCAGAGGATGAGGC
AGAATATTACTGTGTTCTATGGTACAGCAACCGCTGGGTGTTCGGTGGAGGAACCAAACTGACTG
TCCTA 331. MCSP-D5 HL .times. I2C artificial aa
QVQLVQSGAEVKRPGASMKVSCKASGYTFTNYYIHWVRQAPGQGLEWMGWINPNSGATNYAQKFQ
HL
GRVTMTRDTSTSTVYMELSSLRSEDTAVYYCAKSWVSWFASWGQGTLVTVSSGGGGSGGGGSGGG
GSDIVMTQSPDSLAVSLGERATINCKSSQSVLNSKNNRNYLAWYQQKPGQPPKLLIYWASTRESG
VPDRFSGSGSGTDFTLTIDSLQAEDSATYYCQQHYSTPFTFGQGTRLEIKSGGGGSEVQLVESGG
GLVQPGGSLKLSCAASGFTFNKYAMNWVRQAPGKGLEWVARIRSKYNNYATYYADSVKDRFTISR
DDSKNTAYLQMNNLKTEDTAVYYCVRHGNFGNSYISYWAYWGQGTLVTVSSGGGGSGGGGSGGGG
SQTVVTQEPSLTVSPGGTVTLTCGSSTGAVTSGNYPNWVQQKPGQAPRGLIGGTKFLAPGTPARF
SGSLLGGKAALTLSGVQPEDEAEYYCVLWYSNRWVFGGGTKLTVL 332. MCSP-D5 HL
.times. I2C artificial nt
CAGGTGCAGCTGGTCCAGTCTGGGGCTGAGGTGAAGAGGCCTGGGGCCTCAATGAAGGTCTCCTG
HL
CAAGGCTTCTGGGTACACCTTCACCAACTACTATATACACTGGGTGCGACAGGCCCCTGGACAAG
GTCTTGAGTGGATGGGTTGGATCAACCCTAACAGTGGTGCCACAAACTATGCACAGAAGTTCCAG
GGCAGAGTCACCATGACCAGGGACACGTCCACGAGCACAGTCTACATGGAGCTGAGCAGCCTGAG
ATCTGAGGACACGGCCGTGTATTACTGTGCGAAATCCTGGGTCTCCTGGTTTGCTTCCTGGGGTC
AAGGAACCTTGGTCACCGTCTCCTCAGGTGGTGGTGGTTCTGGCGGCGGCGGCTCCGGTGGTGGT
GGTTCTGACATCGTGATGACCCAGTCTCCAGACTCCCTGGCTGTGTCTCTGGGCGAGAGGGCCAC
CATCAACTGCAAGTCCAGCCAGAGTGTTTTAAACAGCAAGAACAATAGGAACTACTTAGCTTGGT
ACCAGCAGAAACCAGGACAGCCTCCTAAGCTGCTCATTTACTGGGCATCTACCCGGGAATCCGGG
GTCCCTGACCGATTCAGTGGCAGCGGGTCTGGGACAGATTTCACTCTCACCATCGATTCCCTGCA
GGCTGAAGATAGTGCAACTTATTACTGTCAGCAACATTATAGTACTCCATTCACTTTTGGCCAGG
GGACCAGACTGGAGATCAAATCCGGAGGTGGTGGATCCGAGGTGCAGCTGGTCGAGTCTGGAGGA
GGATTGGTGCAGCCTGGAGGGTCATTGAAACTCTCATGTGCAGCCTCTGGATTCACCTTCAATAA
GTACGCCATGAACTGGGTCCGCCAGGCTCCAGGAAAGGGTTTGGAATGGGTTGCTCGCATAAGAA
GTAAATATAATAATTATGCAACATATTATGCCGATTCAGTGAAAGACAGGTTCACCATCTCCAGA
GATGATTCAAAAAACACTGCCTATCTACAAATGAACAACTTGAAAACTGAGGACACTGCCGTGTA
CTACTGTGTGAGACATGGGAACTTCGGTAATAGCTACATATCCTACTGGGCTTACTGGGGCCAAG
GGACTCTGGTCACCGTCTCCTCAGGTGGTGGTGGTTCTGGCGGCGGCGGCTCCGGTGGTGGTGGT
TCTCAGACTGTTGTGACTCAGGAACCTTCACTCACCGTATCACCTGGTGGAACAGTCACACTCAC
TTGTGGCTCCTCGACTGGGGCTGTTACATCTGGCAACTACCCAAACTGGGTCCAACAAAAACCAG
GTCAGGCACCCCGTGGTCTAATAGGTGGGACTAAGTTCCTCGCCCCCGGTACTCCTGCCAGATTC
TCAGGCTCCCTGCTTGGAGGCAAGGCTGCCCTCACCCTCTCAGGGGTACAGCCAGAGGATGAGGC
AGAATATTACTGTGTTCTATGGTACAGCAACCGCTGGGTGTTCGGTGGAGGAACCAAACTGACTG
TCCTA 333. MCSP-F7 HL .times. I2C artificial aa
QVQLVQSGAEVKRPGASMKVSCKASGYTFTNYYIHWVRQAPGQGLEWMGWINPNSGATNYAQKFQ
HL
GRVTMTRDTSTSTVYMELSSLRSEDTAVYYCAKSWVSWFASWGQGTLVTVSSGGGGSGGGGSGGG
GSDIVMTQSPDSLAVSLGERATINCKSSQSVLNSKNNRNYLAWYQQKPGQPPKLLIYWASTRESG
VPDRFSGSGSGTDFTLTIDVLQPEDIATYYCQQHYSTPFTFGQGTRLEIKSGGGGSEVQLVESGG
GLVQPGGSLKLSCAASGFTFNKYAMNWVRQAPGKGLEWVARIRSKYNNYATYYADSVKDRFTISR
DDSKNTAYLQMNNLKTEDTAVYYCVRHGNFGNSYISYWAYWGQGTLVTVSSGGGGSGGGGSGGGG
SQTVVTQEPSLTVSPGGTVTLTCGSSTGAVTSGNYPNWVQQKPGQAPRGLIGGTKFLAPGTPARF
SGSLLGGKAALTLSGVQPEDEAEYYCVLWYSNRWVFGGGTKLTVL 334. MCSP-F7 HL
.times. I2C artificial nt
CAGGTGCAGCTGGTCCAGTCTGGGGCTGAGGTGAAGAGGCCTGGGGCCTCAATGAAGGTCTCCTG
HL
CAAGGCTTCTGGGTACACCTTCACCAACTACTATATACACTGGGTGCGACAGGCCCCTGGACAAG
GTCTTGAGTGGATGGGTTGGATCAACCCTAACAGTGGTGCCACAAACTATGCACAGAAGTTCCAG
GGCAGAGTCACCATGACCAGGGACACGTCCACGAGCACAGTCTACATGGAGCTGAGCAGCCTGAG
ATCTGAGGACACGGCCGTGTATTACTGTGCGAAATCCTGGGTCTCCTGGTTTGCTTCCTGGGGTC
AAGGAACCTTGGTCACCGTCTCCTCAGGTGGTGGTGGTTCTGGCGGCGGCGGCTCCGGTGGTGGT
GGTTCTGACATCGTGATGACCCAGTCTCCAGACTCCCTGGCTGTGTCTCTGGGCGAGAGGGCCAC
CATCAACTGCAAGTCCAGCCAGAGTGTTTTAAACAGCAAGAACAATAGGAACTACTTAGCTTGGT
ACCAGCAGAAACCAGGACAGCCTCCTAAGCTGCTCATTTACTGGGCATCTACCCGGGAATCCGGG
GTCCCTGACCGATTCAGTGGCAGCGGGTCTGGGACAGATTTCACTCTCACCATCGATGTCCTGCA
GCCTGAAGATATTGCAACTTATTACTGTCAGCAACATTATAGTACTCCATTCACTTTTGGCCAGG
GGACCAGACTGGAGATCAAATCCGGAGGTGGTGGATCCGAGGTGCAGCTGGTCGAGTCTGGAGGA
GGATTGGTGCAGCCTGGAGGGTCATTGAAACTCTCATGTGCAGCCTCTGGATTCACCTTCAATAA
GTACGCCATGAACTGGGTCCGCCAGGCTCCAGGAAAGGGTTTGGAATGGGTTGCTCGCATAAGAA
GTAAATATAATAATTATGCAACATATTATGCCGATTCAGTGAAAGACAGGTTCACCATCTCCAGA
GATGATTCAAAAAACACTGCCTATCTACAAATGAACAACTTGAAAACTGAGGACACTGCCGTGTA
CTACTGTGTGAGACATGGGAACTTCGGTAATAGCTACATATCCTACTGGGCTTACTGGGGCCAAG
GGACTCTGGTCACCGTCTCCTCAGGTGGTGGTGGTTCTGGCGGCGGCGGCTCCGGTGGTGGTGGT
TCTCAGACTGTTGTGACTCAGGAACCTTCACTCACCGTATCACCTGGTGGAACAGTCACACTCAC
TTGTGGCTCCTCGACTGGGGCTGTTACATCTGGCAACTACCCAAACTGGGTCCAACAAAAACCAG
GTCAGGCACCCCGTGGTCTAATAGGTGGGACTAAGTTCCTCGCCCCCGGTACTCCTGCCAGATTC
TCAGGCTCCCTGCTTGGAGGCAAGGCTGCCCTCACCCTCTCAGGGGTACAGCCAGAGGATGAGGC
AGAATATTACTGTGTTCTATGGTACAGCAACCGCTGGGTGTTCGGTGGAGGAACCAAACTGACTG
TCCTA 335. MCSP-G5 HL .times. I2C artificial aa
QVQLVQSGAEVKRPGASMKVSCKASGYTFTNYYIHWVRQAPGQGLEWMGWINPNSGATNYAQKFQ
HL
GRVTMTRDTSTSTVYMELSSLRSEDTAVYYCAKSWVSWFASWGQGTLVTVSSGGGGSGGGGSGGG
GSDIVMTQSPDSLAVSLGDRATINCKSSQSVLNSKNNRNYLAWYQQKPGQPPKLLIYWASTRESG
VPDRFSGSGSGTDFTLTIDSLQPEDSATYYCQQHYSTPFTFGQGTRLEIKSGGGGSEVQLVESGG
GLVQPGGSLKLSCAASGFTFNKYAMNWVRQAPGKGLEWVARIRSKYNNYATYYADSVKDRFTISR
DDSKNTAYLQMNNLKTEDTAVYYCVRHGNFGNSYISYWAYWGQGTLVTVSSGGGGSGGGGSGGGG
SQTVVTQEPSLTVSPGGTVTLTCGSSTGAVTSGNYPNWVQQKPGQAPRGLIGGTKFLAPGTPARF
SGSLLGGKAALTLSGVQPEDEAEYYCVLWYSNRWVFGGGTKLTVL 336. MCSP-G5 HL
.times. I2C artificial nt
CAGGTGCAGCTGGTCCAGTCTGGGGCTGAGGTGAAGAGGCCTGGGGCCTCAATGAAGGTCTCCTG
HL
CAAGGCTTCTGGGTACACCTTCACCAACTACTATATACACTGGGTGCGACAGGCCCCTGGACAAG
GTCTTGAGTGGATGGGTTGGATCAACCCTAACAGTGGTGCCACAAACTATGCACAGAAGTTCCAG
GGCAGAGTCACCATGACCAGGGACACGTCCACGAGCACAGTCTACATGGAGCTGAGCAGCCTGAG
ATCTGAGGACACGGCCGTGTATTACTGTGCGAAATCCTGGGTCTCCTGGTTTGCTTCCTGGGGTC
AAGGAACCTTGGTCACCGTCTCCTCAGGTGGTGGTGGTTCTGGCGGCGGCGGCTCCGGTGGTGGT
GGTTCTGACATCGTGATGACCCAGTCTCCAGACTCCCTGGCTGTGTCTCTGGGCGACAGGGCCAC
CATCAACTGCAAGTCCAGCCAGAGTGTTTTAAACAGCAAGAACAATAGGAACTACTTAGCTTGGT
ACCAGCAGAAACCAGGACAGCCTCCTAAGCTGCTCATTTACTGGGCATCTACCCGGGAATCCGGG
GTCCCTGACCGATTCAGTGGCAGCGGGTCTGGGACAGATTTCACTCTCACCATCGATTCCCTGCA
GCCTGAGGATAGTGCAACTTATTACTGTCAGCAACATTATAGTACTCCATTCACTTTTGGCCAGG
GGACCAGACTGGAGATCAAATCCGGAGGTGGTGGATCCGAGGTGCAGCTGGTCGAGTCTGGAGGA
GGATTGGTGCAGCCTGGAGGGTCATTGAAACTCTCATGTGCAGCCTCTGGATTCACCTTCAATAA
GTACGCCATGAACTGGGTCCGCCAGGCTCCAGGAAAGGGTTTGGAATGGGTTGCTCGCATAAGAA
GTAAATATAATAATTATGCAACATATTATGCCGATTCAGTGAAAGACAGGTTCACCATCTCCAGA
GATGATTCAAAAAACACTGCCTATCTACAAATGAACAACTTGAAAACTGAGGACACTGCCGTGTA
CTACTGTGTGAGACATGGGAACTTCGGTAATAGCTACATATCCTACTGGGCTTACTGGGGCCAAG
GGACTCTGGTCACCGTCTCCTCAGGTGGTGGTGGTTCTGGCGGCGGCGGCTCCGGTGGTGGTGGT
TCTCAGACTGTTGTGACTCAGGAACCTTCACTCACCGTATCACCTGGTGGAACAGTCACACTCAC
TTGTGGCTCCTCGACTGGGGCTGTTACATCTGGCAACTACCCAAACTGGGTCCAACAAAAACCAG
GTCAGGCACCCCGTGGTCTAATAGGTGGGACTAAGTTCCTCGCCCCCGGTACTCCTGCCAGATTC
TCAGGCTCCCTGCTTGGAGGCAAGGCTGCCCTCACCCTCTCAGGGGTACAGCCAGAGGATGAGGC
AGAATATTACTGTGTTCTATGGTACAGCAACCGCTGGGTGTTCGGTGGAGGAACCAAACTGACTG
TCCTA 337. MCSP-F8 HL .times. I2C artificial aa
QVQLVQSGAEVKRPGASMKVSCKASGYTFTNYYIHWVRQAPGQGLEWMGWINPNSGATNYAQKFQ
HL
GRVTMTRDTSTSTVYMELSSLRSEDTAVYYCAKSWVSWFASWGQGTLVTVSSGGGGSGGGGSGGG
GSDIVMTQSPDSLTVSLGERATINCKSSQSVLNSKNNRNYLAWYQQKPGQPPKLLIYWASTRESG
VPDRFSGSGSGTDFTLTIDSLQAEDSAIYYCQQHYSTPFTFGQGTRLEIKSGGGGSEVQLVESGG
GLVQPGGSLKLSCAASGFTFNKYAMNWVRQAPGKGLEWVARIRSKYNNYATYYADSVKDRFTISR
DDSKNTAYLQMNNLKTEDTAVYYCVRHGNFGNSYISYWAYWGQGTLVTVSSGGGGSGGGGSGGGG
SQTVVTQEPSLTVSPGGTVTLTCGSSTGAVTSGNYPNWVQQKPGQAPRGLIGGTKFLAPGTPARF
SGSLLGGKAALTLSGVQPEDEAEYYCVLWYSNRWVFGGGTKLTVL 338. MCSP-F8 HL
.times. I2C artificial nt
CAGGTGCAGCTGGTCCAGTCTGGGGCTGAGGTGAAGAGGCCTGGGGCCTCAATGAAGGTCTCCTG
HL
CAAGGCTTCTGGGTACACCTTCACCAACTACTATATACACTGGGTGCGACAGGCCCCTGGACAAG
GTCTTGAGTGGATGGGTTGGATCAACCCTAACAGTGGTGCCACAAACTATGCACAGAAGTTCCAG
GGCAGAGTCACCATGACCAGGGACACGTCCACGAGCACAGTCTACATGGAGCTGAGCAGCCTGAG
ATCTGAGGACACGGCCGTGTATTACTGTGCGAAATCCTGGGTCTCCTGGTTTGCTTCCTGGGGTC
AAGGAACCTTGGTCACCGTCTCCTCAGGTGGTGGTGGTTCTGGCGGCGGCGGCTCCGGTGGTGGT
GGTTCTGACATCGTGATGACCCAGTCTCCAGACTCCCTGACTGTGTCTCTGGGCGAGAGGGCCAC
CATCAACTGCAAGTCCAGCCAGAGTGTTTTAAACAGCAAGAACAATAGGAACTACTTAGCTTGGT
ACCAGCAGAAACCAGGACAGCCTCCTAAGCTGCTCATTTACTGGGCATCTACCCGGGAATCCGGG
GTCCCTGACCGATTCAGTGGCAGCGGGTCTGGGACAGATTTCACTCTCACCATCGATTCCCTGCA
GGCTGAAGATAGTGCAATTTATTACTGTCAGCAACATTATAGTACTCCATTCACTTTTGGCCAGG
GGACCAGACTGGAGATCAAATCCGGAGGTGGTGGATCCGAGGTGCAGCTGGTCGAGTCTGGAGGA
GGATTGGTGCAGCCTGGAGGGTCATTGAAACTCTCATGTGCAGCCTCTGGATTCACCTTCAATAA
GTACGCCATGAACTGGGTCCGCCAGGCTCCAGGAAAGGGTTTGGAATGGGTTGCTCGCATAAGAA
GTAAATATAATAATTATGCAACATATTATGCCGATTCAGTGAAAGACAGGTTCACCATCTCCAGA
GATGATTCAAAAAACACTGCCTATCTACAAATGAACAACTTGAAAACTGAGGACACTGCCGTGTA
CTACTGTGTGAGACATGGGAACTTCGGTAATAGCTACATATCCTACTGGGCTTACTGGGGCCAAG
GGACTCTGGTCACCGTCTCCTCAGGTGGTGGTGGTTCTGGCGGCGGCGGCTCCGGTGGTGGTGGT
TCTCAGACTGTTGTGACTCAGGAACCTTCACTCACCGTATCACCTGGTGGAACAGTCACACTCAC
TTGTGGCTCCTCGACTGGGGCTGTTACATCTGGCAACTACCCAAACTGGGTCCAACAAAAACCAG
GTCAGGCACCCCGTGGTCTAATAGGTGGGACTAAGTTCCTCGCCCCCGGTACTCCTGCCAGATTC
TCAGGCTCCCTGCTTGGAGGCAAGGCTGCCCTCACCCTCTCAGGGGTACAGCCAGAGGATGAGGC
AGAATATTACTGTGTTCTATGGTACAGCAACCGCTGGGTGTTCGGTGGAGGAACCAAACTGACTG
TCCTA 339. MCSP-G10 HL .times. I2C artificial aa
QVQLVQSGAEVKRPGASMKVSCKASGYTFTNYYIHWVRQAPGQGLEWMGWINPNSGATNYAQKFQ
HL
GRVTMTRDTSTSTVYMELSSLRSEDTAVYYCAKSWVSWFASWGQGTLVTVSSGGGGSGGGGSGGG
GSDIVMTQSPDSLAVSLGERATINCKSSQSVLSSSNNKNYLNWYQQKPGQPPKLLIYWASTRESG
VPDRFSGSGSGTDFTLTISSLQAEDVAVYYCQQHFNTPFAFGQGTKLEIKSGGGGSEVQLVESGG
GLVQPGGSLKLSCAASGFTFNKYAMNWVRQAPGKGLEWVARIRSKYNNYATYYADSVKDRFTISR
DDSKNTAYLQMNNLKTEDTAVYYCVRHGNFGNSYISYWAYWGQGTLVTVSSGGGGSGGGGSGGGG
SQTVVTQEPSLTVSPGGTVTLTCGSSTGAVTSGNYPNWVQQKPGQAPRGLIGGTKFLAPGTPARF
SGSLLGGKAALTLSGVQPEDEAEYYCVLWYSNRWVFGGGTKLTVL 340. MCSP-G10 HL
.times. I2C artificial nt
CAGGTGCAGCTGGTCCAGTCTGGGGCTGAGGTGAAGAGGCCTGGGGCCTCAATGAAGGTCTCCTG
HL
CAAGGCTTCTGGGTACACCTTCACCAACTACTATATACACTGGGTGCGACAGGCCCCTGGACAAG
GTCTTGAGTGGATGGGTTGGATCAACCCTAACAGTGGTGCCACAAACTATGCACAGAAGTTCCAG
GGCAGAGTCACCATGACCAGGGACACGTCCACGAGCACAGTCTACATGGAGCTGAGCAGCCTGAG
ATCTGAGGACACGGCCGTGTATTACTGTGCGAAATCCTGGGTCTCCTGGTTTGCTTCCTGGGGTC
AAGGAACCTTGGTCACCGTCTCCTCAGGTGGTGGTGGTTCTGGCGGCGGCGGCTCCGGTGGTGGT
GGTTCTGACATCGTGATGACACAGTCTCCAGACTCCCTGGCTGTGTCTCTGGGCGAGAGGGCCAC
CATCAACTGCAAGTCCAGCCAGAGTGTCTTATCCAGCTCCAACAATAAGAACTACTTAAATTGGT
ACCAGCAGAAACCAGGACAGCCTCCTAAGTTGCTCATTTACTGGGCATCTACCCGGGAATCCGGG
GTCCCTGACCGATTCAGTGGCAGCGGGTCTGGGACAGATTTCACTCTCACCATCAGCAGCCTGCA
GGCTGAAGATGTGGCGGTTTATTACTGTCAACAACATTTTAATACTCCGTTCGCTTTTGGCCAGG
GGACCAAGCTGGAGATCAAATCCGGAGGTGGTGGATCCGAGGTGCAGCTGGTCGAGTCTGGAGGA
GGATTGGTGCAGCCTGGAGGGTCATTGAAACTCTCATGTGCAGCCTCTGGATTCACCTTCAATAA
GTACGCCATGAACTGGGTCCGCCAGGCTCCAGGAAAGGGTTTGGAATGGGTTGCTCGCATAAGAA
GTAAATATAATAATTATGCAACATATTATGCCGATTCAGTGAAAGACAGGTTCACCATCTCCAGA
GATGATTCAAAAAACACTGCCTATCTACAAATGAACAACTTGAAAACTGAGGACACTGCCGTGTA
CTACTGTGTGAGACATGGGAACTTCGGTAATAGCTACATATCCTACTGGGCTTACTGGGGCCAAG
GGACTCTGGTCACCGTCTCCTCAGGTGGTGGTGGTTCTGGCGGCGGCGGCTCCGGTGGTGGTGGT
TCTCAGACTGTTGTGACTCAGGAACCTTCACTCACCGTATCACCTGGTGGAACAGTCACACTCAC
TTGTGGCTCCTCGACTGGGGCTGTTACATCTGGCAACTACCCAAACTGGGTCCAACAAAAACCAG
GTCAGGCACCCCGTGGTCTAATAGGTGGGACTAAGTTCCTCGCCCCCGGTACTCCTGCCAGATTC
TCAGGCTCCCTGCTTGGAGGCAAGGCTGCCCTCACCCTCTCAGGGGTACAGCCAGAGGATGAGGC
AGAATATTACTGTGTTCTATGGTACAGCAACCGCTGGGTGTTCGGTGGAGGAACCAAACTGACTG
TCCTA 341. Human CD3.epsilon. 1-8 human aa QDGNEEMG (N-terminus)
342. Saimiri sciureus Saimiri aa QDGNEEIG CD3.epsilon. 1-8 sciureus
(N-terminus) 343. Thioate-modified artificial nt
TCCATGACGTTCCTGATGCT CpG-Oligonucleotide 344. MVH1 artificial nt
(GC)AGGTGCAGCTCGAGGAGTCAGGACCT 345. MVH2 artificial nt
GAGGTCCAGCTCGAGCAGTCTGGACCT 346. MVH3 artificial nt
CAGGTCCAACTCGAGCAGCCTGGGGCT 347. MVH4 artificial nt
GAGGTTCAGCTCGAGCAGTCTGGGGCA 348. MVH5 artificial nt
GA(AG)GTGAAGCTCGAGGAGTCTGGAGGA 349. MVH6 artificial nt
GAGGTGAAGCTTCTCGAGTCTGGAGGT 350. MVH7 artificial nt
GAAGTGAAGCTCGAGGAGTCTGGGGGA 351. MVH8 artificial nt
GAGGTTCAGCTCGAGCAGTCTGGAGCT 352. MuVHBstEII artificial nt
TGAGGAGACGGTGACCGTGGTCCCTTGGCCCCAG 353. MUVK1 artificial nt
CCAGTTCCGAGCTCGTTGTGACTCAGGAATCT 354. MUVK2 artificial nt
CCAGTTCCGAGCTCGTGTTGACGCAGCCGCCC 355. MUVK3 artificial nt
CCAGTTCCGAGCTCGTGCTCACCCAGTCTCCA 356. MUVK4 artificial nt
CCAGTTCCGAGCTCCAGATGACCCAGTCTCCA 357. MUVK5 artificial nt
CCAGATGTGAGCTCGTGATGACCCAGACTCCA 358. MUVK6 artificial nt
CCAGATGTGAGCTCGTCATGACCCAGTCTCCA 359. MUVK7 artificial nt
CCAGTTCCGAGCTCGTGATGACACAGTCTCCA 360. MuVkHindIII/BsiW1 artificial
nt TGGTGCACTAGTCGTACGTTTGATCTCAAGCTTGGTCCC 361. forward primer
artificial nt GATCTGGTCTACACCATCGAGC 362. reverse primer artificial
nt GGAGCTGCTGCTGGCTCAGTGAGG 363. forward primer artificial nt
TTCCAGCTGAGCATGTCTGATGG 364. reverse primer artificial nt
CGATCAGCATCTGGGCCCAGG 365. forward primer artificial nt
GTGGAGCAGTTCACTCAGCAGGACC 366. reverse primer artificial nt
GCCTTCACACCCAGTACTGGCC 367. forward primer artificial nt
TCCCGTACGAGATCTGGATCCCAATTGGATGGCGGACTCGTGCTGTTCTCACACAGAGG 368.
reverse primer artificial nt
AGTGGGTCGACTCACACCCAGTACTGGCCATTCTTAAGGGCAGGG 369. forward primer
artificial nt
GAGGAATTCACCATGCCGCTGCTGCTACTGCTGCCCCTGCTGTGGGCAGGGGCCCTGGCTATGG
370. reverse primer artificial nt GATTTGTAACTGTATTTGGTACTTCC 371.
forward primer artificial nt ATTCCGCCTCCTTGGGGATCC 372. reverse
primer artificial nt GCATAGGAGACATTGAGCTGGATGG 373. forward primer
artificial nt GCACCAACCTGACCTGTCAGG 374. reverse primer artificial
nt AGTGGGTCGACTCACTGGGTCCTGACCTCTGAGTATTCG 375. forward primer
artificial nt CACTGTGGCCCAGGTTCGAGG 376. reverse primer artificial
nt GACATACCACACAAATTCAATACGG 377. forward primer artificial nt
GCTCTGCTCGCGCCGAGATGTGG 378. reverse primer artificial nt
ACGCTGGACACCACCTCCAGG 379. forward primer artificial nt
GGTTCTACTGAGTGGGCAGAGG
380. reverse primer artificial nt ACTTGTTGTGGCTGCTTGGAGC 381.
forward primer artificial nt GGGTGAAGTCCTATCCAGATGG 382. reverse
primer artificial nt GTGCTCTGCCTGAAGCAATTCC 383. forward primer
artificial nt CTCGGCTTCCTCTTCGGGTGG 384. reverse primer artificial
nt GCATATTCATTTGCTGGGTAACCTGG 385. macaque PSMA artificial nt
ATGTGGAATCTCCTGCACGAAACCGACTCGGCTGTGGCCACCGCGCGCCGCCCGCGCTGGCTGTG
(Cynomolgus)
CGCTGGGGCACTGGTGCTGGCGGGTGGCTTCTTTCTCCTCGGCTTCCTCTTCGGATGGTTTATAA
AATCCTCCAGTGAAGCTACTAACATTACTCCAAAGCATAATATGAAAGCATTTTTGGATGAACTG
AAAGCTGAGAACATCAAGAAGTTCTTACATAATTTTACACAGATACCACATTTAGCAGGAACAGA
ACAAAACTTTCAACTTGCAAAGCAAATTCAATCCCAGTGGAAAGAATTTGGCCTGGATTCTGTTG
AGCTAACTCATTATGATGTCCTGTTGTCCTACCCAAATAAGACTCATCCCAACTACATCTCAATA
ATTAATGAAGATGGAAATGAGATTTTCAACACATCATTATTTGAACCACCTCCTGCAGGATATGA
AAATGTTTCGGATATTGTACCACCTTTCAGTGCTTTCTCTCCTCAAGGAATGCCAGAGGGCGATC
TAGTGTATGTTAACTATGCACGAACTGAAGACTTCTTTAAATTGGAACGGGACATGAAAATCAAT
TGCTCTGGGAAAATTGTAATTGCCAGATATGGGAAAGTTTTCAGAGGAAATAAGGTTAAAAATGC
CCAGCTGGCAGGGGCCACAGGAGTCATTCTCTACTCAGACCCTGCTGACTACTTTGCTCCTGGGG
TAAAGTCTTATCCAGATGGTTGGAATCTTCCTGGAGGTGGTGTCCAGCGTGGAAATATCCTAAAT
CTGAATGGTGCAGGAGACCCTCTCACACCAGGTTACCCAGCAAATGAATATGCTTATAGGCGTGG
AATGGCAGAGGCTGTTGGTCTTCCAAGTATTCCCGTTCATCCAATTGGGTACTATGATGCACAGA
AGCTCCTAGAAAAAATGGGTGGCTCAGCATCACCAGATAGCAGCTGGAGAGGAAGTCTCAAAGTG
CCCTACAATGTTGGACCTGGCTTTACTGGAAACTTTTCTACACAAAAAGTCAAGATGCACATCCA
CTCTACCAGTGAAGTGACAAGAATTTACAATGTGATAGGTACTCTCAGAGGAGCAGTGGAACCAG
ACAGATACGTCATTCTGGGAGGTCACCGGGACTCATGGGTGTTTGGTGGTATTGACCCTCAGAGT
GGAGCAGCTGTTGTTCATGAAATTGTGAGGAGCTTTGGAACGCTGAAAAAGGAAGGGTGGAGACC
TAGAAGAACAATTTTGTTTGCAAGCTGGGATGCAGAAGAATTTGGTCTTCTTGGTTCTACTGAAT
GGGCAGAGGAGAATTCAAGACTCCTTCAAGAGCGTGGCGTGGCTTATATTAATGCTGATTCGTCT
ATAGAGGGAAACTACACTCTGAGAGTTGATTGTACACCACTGATGTACAGCTTGGTATACAACCT
AACAAAAGAGCTGGAAAGCCCTGATGAAGGCTTTGAAGGCAAATCTCTTTATGAAAGTTGGACTA
AAAAAAGTCCTTCCCCCGAGTTCAGTGGCATGCCCAGGATAAGCAAATTGGGATCTGGAAATGAT
TTTGAGGTGTTCTTCCAACGACTTGGAATTGCCTCAGGCAGAGCACGGTATACTAAAAATTGGGA
AACAAACAAATTCAGCAGCTATCCACTGTATCACAGTGTCTATGAGACATATGAGTTGGTGGAAA
AGTTTTATGATCCAATGTTTAAATATCACCTCACTGTGGCCCAGGTTCGAGGAGGGATGGTGTTT
GAACTAGCCAATTCCGTAGTGCTCCCTTTTGATTGTCGAGATTATGCTGTAGTTTTAAGAAAGTA
TGCTGACAAAATCTACAATATTTCTATGAAACATCCACAGGAAATGAAGACATACAGTGTATCAT
TTGATTCACTTTTTTCTGCAGTAAAGAATTTTACAGAAATTGCTTCCAAGTTCAGCGAGAGACTC
CGGGACTTTGACAAAAGCAACCCAATATTATTAAGAATGATGAATGATCAACTCATGTTTCTGGA
AAGAGCATTTATTGATCCATTAGGGTTACCAGACAGACCTTTTTATAGGCATGTCATCTATGCTC
CAAGCAGCCACAACAAGTATGCAGGGGAGTCATTCCCAGGAATTTATGATGCTCTGTTTGATATC
GAAAGCAAAGTGGACCCTTCCCAGGCCTGGGGAGAAGTGAAGAGACAGATTTCTGTTGCAACCTT
CACAGTGCAAGCAGCTGCAGAGACTTTGAGTGAAGTGGCCTAA 386. macaque PSMA
artificial aa
MWNLLHETDSAVATARRPRWLCAGALVLAGGFFLLGFLFGWFIKSSSEATNITPKHNMKAFLDEL
(Cynomolgus)
KAENIKKFLHNFTQIPHLAGTEQNFQLAKQIQSQWKEFGLDSVELTHYDVLLSYPNKTHPNYISI
INEDGNEIFNTSLFEPPPAGYENVSDIVPPFSAFSPQGMPEGDLVYVNYARTEDFFKLERDMKIN
CSGKIVIARYGKVFRGNKVKNAQLAGATGVILYSDPADYFAPGVKSYPDGWNLPGGGVQRGNILN
LNGAGDPLTPGYPANEYAYRRGMAEAVGLPSIPVHPIGYYDAQKLLEKMGGSASPDSSWRGSLKV
PYNVGPGFTGNFSTQKVKMHIHSTSEVTRIYNVIGTLRGAVEPDRYVILGGHRDSWVFGGIDPQS
GAAVVHEIVRSFGTLKKEGWRPRRTILFASWDAEEFGLLGSTEWAEENSRLLQERGVAYINADSS
IEGNYTLRVDCTPLMYSLVYNLTKELESPDEGFEGKSLYESWTKKSPSPEFSGMPRISKLGSGND
FEVFFQRLGIASGRARYTKNWETNKFSSYPLYHSVYETYELVEKFYDPMFKYHLTVAQVRGGMVF
ELANSVVLPFDCRDYAVVLRKYADKIYNISMKHPQEMKTYSVSFDSLFSAVKNFTEIASKFSERL
RDFDKSNPILLRMMNDQLMFLERAFIDPLGLPDRPFYRHVIYAPSSHNKYAGESFPGIYDALFDI
ESKVDPSQAWGEVKRQISVATFTVQAAAETLSEVA 387. PSMA-3 L artificial aa
DIQMTQSPKFMSTSVGDRVSVTCKASQNVDTNVAWYQQKPGQSPKALIYSASYRYSDVPDRFTGS
ESGTDFTLTISNVQSEDLAEYFCQQYDSYPYTFGGGTKLEIK 388. PSMA-3 L artificial
nt
GACATCCAGATGACCCAGTCCCCCAAGTTCATGTCCACCTCCGTGGGCGACAGAGTGTCCGTGAC
CTGCAAGGCCTCCCAGAACGTGGACACCAACGTGGCCTGGTATCAGCAGAAGCCCGGCCAGTCCC
CTAAGGCCCTGATCTACTCCGCCTCCTACCGGTACTCTGACGTGCCTGACCGGTTCACCGGCTCC
GAGTCCGGCACCGACTTCACCCTGACCATCTCCAACGTGCAGTCTGAGGACCTGGCCGAGTACTT
CTGCCAGCAGTACGACTCCTACCCTTACACCTTCGGCGGAGGGACCAAGCTGGAAATCAAG 389.
PSMA-3 LCDR1 artificial aa KASQNVDTNVA 390. PSMA-3 LCDR2 artificial
aa SASYRYS 391. PSMA-3 LCDR3 artificial aa QQYDSYPYT 392. PSMA-3 H
artificial aa
DVKLVESGGGLVKPGESLKLSCIASGFTFSDYYMYWVRQTPEKRLEWVAIISDGGYYTYYSDIIK
GRFTISRDNAKNNLYLQMSSLKSEDTAMYYCTRGFPLLRHGAMDYWGLGTSVTVSS 393.
PSMA-3 H artificial nt
GACGTGAAACTGGTGGAGTCTGGCGGCGGACTGGTGAAGCCTGGCGAGTCCCTGAAGCTGTCCTG
TATCGCCTCCGGCTTCACCTTCTCCGACTACTACATGTACTGGGTGCGCCAGACCCCTGAGAAGC
GGCTGGAATGGGTGGCCATCATCTCCGACGGCGGCTACTACACCTACTACTCCGACATCATCAAG
GGCCGGTTCACCATCTCCCGGGACAACGCCAAGAACAACCTGTACCTGCAGATGTCCTCCCTGAA
GTCCGAGGACACCGCCATGTACTACTGCACCCGGGGCTTCCCTCTGCTGAGACACGGCGCCATGG
ATTACTGGGGCCTGGGCACCTCTGTGACCGTGTCCTCT 394. PSMA-3 HCDR1 artificial
aa DYYMY 395. PSMA-3 HCDR2 artificial aa IISDGGYYTYYSDIIKG 396.
PSMA-3 HCDR3 artificial aa GFPLLRHGAMDY 397. PSMA-3 HL artificial
aa
DVKLVESGGGLVKPGESLKLSCIASGFTFSDYYMYWVRQTPEKRLEWVAIISDGGYYTYYSDIIK
GRFTISRDNAKNNLYLQMSSLKSEDTAMYYCTRGFPLLRHGAMDYWGLGTSVTVSSGGGGSGGGG
SGGGGSDIQMTQSPKFMSTSVGDRVSVTCKASQNVDTNVAWYQQKPGQSPKALIYSASYRYSDVP
DRFTGSESGTDFTLTISNVQSEDLAEYFCQQYDSYPYTFGGGTKLEIK 398. PSMA-3 HL
artificial nt
GACGTGAAACTGGTGGAGTCTGGCGGCGGACTGGTGAAGCCTGGCGAGTCCCTGAAGCTGTCCTG
TATCGCCTCCGGCTTCACCTTCTCCGACTACTACATGTACTGGGTGCGCCAGACCCCTGAGAAGC
GGCTGGAATGGGTGGCCATCATCTCCGACGGCGGCTACTACACCTACTACTCCGACATCATCAAG
GGCCGGTTCACCATCTCCCGGGACAACGCCAAGAACAACCTGTACCTGCAGATGTCCTCCCTGAA
GTCCGAGGACACCGCCATGTACTACTGCACCCGGGGCTTCCCTCTGCTGAGACACGGCGCCATGG
ATTACTGGGGCCTGGGCACCTCTGTGACCGTGTCCTCTGGCGGAGGGGGCAGTGGAGGCGGAGGA
AGTGGAGGGGGCGGATCCGACATCCAGATGACCCAGTCCCCCAAGTTCATGTCCACCTCCGTGGG
CGACAGAGTGTCCGTGACCTGCAAGGCCTCCCAGAACGTGGACACCAACGTGGCCTGGTATCAGC
AGAAGCCCGGCCAGTCCCCTAAGGCCCTGATCTACTCCGCCTCCTACCGGTACTCTGACGTGCCT
GACCGGTTCACCGGCTCCGAGTCCGGCACCGACTTCACCCTGACCATCTCCAACGTGCAGTCTGA
GGACCTGGCCGAGTACTTCTGCCAGCAGTACGACTCCTACCCTTACACCTTCGGCGGAGGGACCA
AGCTGGAAATCAAG 399. PSMA-3 HL .times. I2C artificial aa
DVKLVESGGGLVKPGESLKLSCIASGFTFSDYYMYWVRQTPEKRLEWVAIISDGGYYTYYSDIIK
HL
GRFTISRDNAKNNLYLQMSSLKSEDTAMYYCTRGFPLLRHGAMDYWGLGTSVTVSSGGGGSGGGG
SGGGGSDIQMTQSPKFMSTSVGDRVSVTCKASQNVDTNVAWYQQKPGQSPKALIYSASYRYSDVP
DRFTGSESGTDFTLTISNVQSEDLAEYFCQQYDSYPYTFGGGTKLEIKSGGGGSEVQLVESGGGL
VQPGGSLKLSCAASGFTFNKYAMNWVRQAPGKGLEWVARIRSKYNNYATYYADSVKDRFTISRDD
SKNTAYLQMNNLKTEDTAVYYCVRHGNFGNSYISYWAYWGQGTLVTVSSGGGGSGGGGSGGGGSQ
TVVTQEPSLTVSPGGTVTLTCGSSTGAVTSGNYPNWVQQKPGQAPRGLIGGTKFLAPGTPARFSG
SLLGGKAALTLSGVQPEDEAEYYCVLWYSNRWVFGGGTKLTVL 400. PSMA-3 HL .times.
I2C artificial nt
GACGTGAAACTGGTGGAGTCTGGCGGCGGACTGGTGAAGCCTGGCGAGTCCCTGAAGCTGTCCTG
HL
TATCGCCTCCGGCTTCACCTTCTCCGACTACTACATGTACTGGGTGCGCCAGACCCCTGAGAAGC
GGCTGGAATGGGTGGCCATCATCTCCGACGGCGGCTACTACACCTACTACTCCGACATCATCAAG
GGCCGGTTCACCATCTCCCGGGACAACGCCAAGAACAACCTGTACCTGCAGATGTCCTCCCTGAA
GTCCGAGGACACCGCCATGTACTACTGCACCCGGGGCTTCCCTCTGCTGAGACACGGCGCCATGG
ATTACTGGGGCCTGGGCACCTCTGTGACCGTGTCCTCTGGCGGAGGGGGCAGTGGAGGCGGAGGA
AGTGGAGGGGGCGGATCCGACATCCAGATGACCCAGTCCCCCAAGTTCATGTCCACCTCCGTGGG
CGACAGAGTGTCCGTGACCTGCAAGGCCTCCCAGAACGTGGACACCAACGTGGCCTGGTATCAGC
AGAAGCCCGGCCAGTCCCCTAAGGCCCTGATCTACTCCGCCTCCTACCGGTACTCTGACGTGCCT
GACCGGTTCACCGGCTCCGAGTCCGGCACCGACTTCACCCTGACCATCTCCAACGTGCAGTCTGA
GGACCTGGCCGAGTACTTCTGCCAGCAGTACGACTCCTACCCTTACACCTTCGGCGGAGGGACCA
AGCTGGAAATCAAGTCCGGAGGTGGTGGATCCGAGGTGCAGCTGGTCGAGTCTGGAGGAGGATTG
GTGCAGCCTGGAGGGTCATTGAAACTCTCATGTGCAGCCTCTGGATTCACCTTCAATAAGTACGC
CATGAACTGGGTCCGCCAGGCTCCAGGAAAGGGTTTGGAATGGGTTGCTCGCATAAGAAGTAAAT
ATAATAATTATGCAACATATTATGCCGATTCAGTGAAAGACAGGTTCACCATCTCCAGAGATGAT
TCAAAAAACACTGCCTATCTACAAATGAACAACTTGAAAACTGAGGACACTGCCGTGTACTACTG
TGTGAGACATGGGAACTTCGGTAATAGCTACATATCCTACTGGGCTTACTGGGGCCAAGGGACTC
TGGTCACCGTCTCCTCAGGTGGTGGTGGTTCTGGCGGCGGCGGCTCCGGTGGTGGTGGTTCTCAG
ACTGTTGTGACTCAGGAACCTTCACTCACCGTATCACCTGGTGGAACAGTCACACTCACTTGTGG
CTCCTCGACTGGGGCTGTTACATCTGGCAACTACCCAAACTGGGTCCAACAAAAACCAGGTCAGG
CACCCCGTGGTCTAATAGGTGGGACTAAGTTCCTCGCCCCCGGTACTCCTGCCAGATTCTCAGGC
TCCCTGCTTGGAGGCAAGGCTGCCCTCACCCTCTCAGGGGTACAGCCAGAGGATGAGGCAGAATA
TTACTGTGTTCTATGGTACAGCAACCGCTGGGTGTTCGGTGGAGGAACCAAACTGACTGTCCTA
401. PSMA-4 L artificial aa
DIELTQSPLSLPVILGDQASISCRSSQSLVHSNGNTYLHWFLQKPGQSPKLLIYTVSNRFSGVPD
RFSGSGSGTDFTLKISRVEAEDLGVYFCSQSTHVPTFGGGTKLEIK 402. PSMA-4 L
artificial nt
GACATCGAGCTGACCCAGTCCCCCCTGTCCCTGCCTGTGATCCTGGGCGACCAGGCCTCCATCTC
CTGCCGGTCCTCCCAGTCCCTGGTGCACTCCAACGGCAATACCTACCTGCACTGGTTTCTGCAGA
AGCCTGGCCAGTCCCCTAAGCTGCTGATCTACACCGTGTCCAACCGGTTCTCCGGCGTGCCTGAC
AGGTTCTCTGGCTCCGGCTCCGGCACCGACTTCACCCTGAAGATCTCCCGGGTGGAGGCCGAAGA
TCTGGGCGTGTACTTTTGCTCCCAGTCCACCCACGTGCCTACCTTCGGCGGAGGGACCAAGCTGG
AAATCAAG 403. PSMA-4 LCDR1 artificial aa RSSQSLVHSNGNTYLH 404.
PSMA-4 LCDR2 artificial aa TVSNRFS 405. PSMA-4 LCDR3 artificial aa
SQSTHVPT 406. PSMA-4 H artificial aa
QVQLQQSGAELVEPGASVKLSCKASGYTFTYFDINWLRQRPEQGLEWIGGISPGDGNTNYNENFK
GKATLTIDKSSTTAYIQLSRLTSEDSAVYFCARDGNFPYYAMDSWGQGTSVTVSS 407. PSMA-4
H artificial nt
CAGGTGCAGCTGCAGCAGTCTGGCGCCGAACTGGTGGAGCCTGGCGCCTCCGTGAAGCTGTCCTG
CAAGGCCTCCGGCTACACCTTCACCTACTTCGACATCAACTGGCTGCGGCAGAGGCCTGAGCAGG
GCCTGGAATGGATCGGCGGCATCTCCCCTGGCGACGGCAACACCAACTACAACGAGAACTTCAAG
GGCAAGGCCACCCTGACCATCGACAAGTCCTCCACCACCGCCTACATCCAGCTGTCCCGGCTGAC
CTCTGAGGACTCCGCCGTGTACTTCTGCGCCAGGGACGGCAACTTCCCTTACTACGCCATGGACT
CTTGGGGCCAGGGCACCTCCGTGACCGTGTCTAGT 408. PSMA-4 HCDR1 artificial aa
YFDIN 409. PSMA-4 HCDR2 artificial aa GISPGDGNTNYNENFKG 410. PSMA-4
HCDR3 artificial aa DGNFPYYAMDS 411. PSMA-4 HL artificial aa
QVQLQQSGAELVEPGASVKLSCKASGYTFTYFDINWLRQRPEQGLEWIGGISPGDGNTNYNENFK
GKATLTIDKSSTTAYIQLSRLTSEDSAVYFCARDGNFPYYAMDSWGQGTSVTVSSGGGGSGGGGS
GGGGSDIELTQSPLSLPVILGDQASISCRSSQSLVHSNGNTYLHWFLQKPGQSPKLLIYTVSNRF
SGVPDRFSGSGSGTDFTLKISRVEAEDLGVYFCSQSTHVPTFGGGTKLEIK 412. PSMA-4 HL
artificial nt
CAGGTGCAGCTGCAGCAGTCTGGCGCCGAACTGGTGGAGCCTGGCGCCTCCGTGAAGCTGTCCTG
CAAGGCCTCCGGCTACACCTTCACCTACTTCGACATCAACTGGCTGCGGCAGAGGCCTGAGCAGG
GCCTGGAATGGATCGGCGGCATCTCCCCTGGCGACGGCAACACCAACTACAACGAGAACTTCAAG
GGCAAGGCCACCCTGACCATCGACAAGTCCTCCACCACCGCCTACATCCAGCTGTCCCGGCTGAC
CTCTGAGGACTCCGCCGTGTACTTCTGCGCCAGGGACGGCAACTTCCCTTACTACGCCATGGACT
CTTGGGGCCAGGGCACCTCCGTGACCGTGTCTAGTGGCGGCGGAGGATCTGGCGGAGGGGGATCT
GGGGGCGGAGGAAGCGACATCGAGCTGACCCAGTCCCCCCTGTCCCTGCCTGTGATCCTGGGCGA
CCAGGCCTCCATCTCCTGCCGGTCCTCCCAGTCCCTGGTGCACTCCAACGGCAATACCTACCTGC
ACTGGTTTCTGCAGAAGCCTGGCCAGTCCCCTAAGCTGCTGATCTACACCGTGTCCAACCGGTTC
TCCGGCGTGCCTGACAGGTTCTCTGGCTCCGGCTCCGGCACCGACTTCACCCTGAAGATCTCCCG
GGTGGAGGCCGAAGATCTGGGCGTGTACTTTTGCTCCCAGTCCACCCACGTGCCTACCTTCGGCG
GAGGGACCAAGCTGGAAATCAAG 413. PSMA-4HL .times. I2C HL artificial aa
QVQLQQSGAELVEPGASVKLSCKASGYTFTYFDINWLRQRPEQGLEWIGGISPGDGNTNYNENFK
GKATLTIDKSSTTAYIQLSRLTSEDSAVYFCARDGNFPYYAMDSWGQGTSVTVSSGGGGSGGGGS
GGGGSDIELTQSPLSLPVILGDQASISCRSSQSLVHSNGNTYLHWFLQKPGQSPKLLIYTVSNRF
SGVPDRFSGSGSGTDFTLKISRVEAEDLGVYFCSQSTHVPTFGGGTKLEIKSGGGGSEVQLVESG
GGLVQPGGSLKLSCAASGFTFNKYAMNWVRQAPGKGLEWVARIRSKYNNYATYYADSVKDRFTIS
RDDSKNTAYLQMNNLKTEDTAVYYCVRHGNFGNSYISYWAYWGQGTLVTVSSGGGGSGGGGSGGG
GSQTVVTQEPSLTVSPGGTVTLTCGSSTGAVTSGNYPNWVQQKPGQAPRGLIGGTKFLAPGTPAR
FSGSLLGGKAALTLSGVQPEDEAEYYCVLWYSNRWVFGGGTKLTVL 414. PSMA-4HL
.times. I2C HL artificial nt
CAGGTGCAGCTGCAGCAGTCTGGCGCCGAACTGGTGGAGCCTGGCGCCTCCGTGAAGCTGTCCTG
CAAGGCCTCCGGCTACACCTTCACCTACTTCGACATCAACTGGCTGCGGCAGAGGCCTGAGCAGG
GCCTGGAATGGATCGGCGGCATCTCCCCTGGCGACGGCAACACCAACTACAACGAGAACTTCAAG
GGCAAGGCCACCCTGACCATCGACAAGTCCTCCACCACCGCCTACATCCAGCTGTCCCGGCTGAC
CTCTGAGGACTCCGCCGTGTACTTCTGCGCCAGGGACGGCAACTTCCCTTACTACGCCATGGACT
CTTGGGGCCAGGGCACCTCCGTGACCGTGTCTAGTGGCGGCGGAGGATCTGGCGGAGGGGGATCT
GGGGGCGGAGGAAGCGACATCGAGCTGACCCAGTCCCCCCTGTCCCTGCCTGTGATCCTGGGCGA
CCAGGCCTCCATCTCCTGCCGGTCCTCCCAGTCCCTGGTGCACTCCAACGGCAATACCTACCTGC
ACTGGTTTCTGCAGAAGCCTGGCCAGTCCCCTAAGCTGCTGATCTACACCGTGTCCAACCGGTTC
TCCGGCGTGCCTGACAGGTTCTCTGGCTCCGGCTCCGGCACCGACTTCACCCTGAAGATCTCCCG
GGTGGAGGCCGAAGATCTGGGCGTGTACTTTTGCTCCCAGTCCACCCACGTGCCTACCTTCGGCG
GAGGGACCAAGCTGGAAATCAAGTCCGGAGGTGGTGGATCCGAGGTGCAGCTGGTCGAGTCTGGA
GGAGGATTGGTGCAGCCTGGAGGGTCATTGAAACTCTCATGTGCAGCCTCTGGATTCACCTTCAA
TAAGTACGCCATGAACTGGGTCCGCCAGGCTCCAGGAAAGGGTTTGGAATGGGTTGCTCGCATAA
GAAGTAAATATAATAATTATGCAACATATTATGCCGATTCAGTGAAAGACAGGTTCACCATCTCC
AGAGATGATTCAAAAAACACTGCCTATCTACAAATGAACAACTTGAAAACTGAGGACACTGCCGT
GTACTACTGTGTGAGACATGGGAACTTCGGTAATAGCTACATATCCTACTGGGCTTACTGGGGCC
AAGGGACTCTGGTCACCGTCTCCTCAGGTGGTGGTGGTTCTGGCGGCGGCGGCTCCGGTGGTGGT
GGTTCTCAGACTGTTGTGACTCAGGAACCTTCACTCACCGTATCACCTGGTGGAACAGTCACACT
CACTTGTGGCTCCTCGACTGGGGCTGTTACATCTGGCAACTACCCAAACTGGGTCCAACAAAAAC
CAGGTCAGGCACCCCGTGGTCTAATAGGTGGGACTAAGTTCCTCGCCCCCGGTACTCCTGCCAGA
TTCTCAGGCTCCCTGCTTGGAGGCAAGGCTGCCCTCACCCTCTCAGGGGTACAGCCAGAGGATGA
GGCAGAATATTACTGTGTTCTATGGTACAGCAACCGCTGGGTGTTCGGTGGAGGAACCAAACTGA
CTGTCCTA 415. PSMA-6 L artificial aa
DIKMTQSPSSMYASLGERVTITCKASQDIYSYLIWFQQKPGKSPKTLIYRANRLVDGVPSRFSGS
GSGQDYSLTISSLEYEDMGIYYCLQYDEFATFGSGTKLEMK 416. PSMA-6 L artificial
nt
GACATCAAGATGACCCAGTCCCCCTCCTCCATGTACGCCTCCCTGGGCGAGAGAGTGACCATCAC
CTGCAAGGCCTCCCAGGACATCTACTCCTACCTGATCTGGTTCCAGCAGAAGCCTGGCAAGTCCC
CTAAGACCCTGATCTACCGGGCCAACAGACTGGTGGACGGCGTGCCTTCCAGGTTCTCCGGCTCC
GGCTCTGGCCAGGACTACTCCCTGACCATCTCCTCCCTGGAATACGAGGACATGGGCATCTACTA
CTGCCTGCAGTACGACGAGTTCGCCACCTTCGGCTCCGGCACCAAGCTGGAAATGAAG 417.
PSMA-6 LCDR1 artificial aa KASQDIYSYLI 418. PSMA-6 LCDR2 artificial
aa RANRLVD 419. PSMA-6 LCDR3 artificial aa LQYDEFAT 420. PSMA-6 H
artificial aa
DVHLQESGPGLVKPSQSLSLTCTVTGYSITSDYAWNWIRQFPGNKLEWMGYISFSGSTSYNPSLK
SRISVTRDTSKNQFFLQLNSVTTEDTATYYCARWNYYGSSHVWFAYWGQGTLVTVSS 421.
PSMA-6 H artificial nt
GACGTGCACCTGCAGGAATCTGGCCCTGGCCTGGTGAAGCCTTCCCAGTCCCTGTCCCTGACCTG
CACCGTGACCGGCTACTCCATCACCTCCGACTACGCCTGGAACTGGATCCGGCAGTTCCCTGGCA
ATAAGCTGGAATGGATGGGCTACATCTCCTTCTCCGGCAGCACCTCCTACAACCCTTCCCTGAAG
TCCCGGATCTCCGTGACCCGGGACACCTCCAAGAACCAGTTCTTCCTGCAGCTGAACTCCGTGAC
CACCGAGGACACCGCCACCTACTACTGCGCCCGGTGGAACTACTACGGCTCCTCCCACGTGTGGT
TCGCTTACTGGGGCCAGGGCACCCTGGTGACCGTGTCCTCC 422. PSMA-6 HCDR1
artificial aa SDYAWN 423. PSMA-6 HCDR2 artificial aa
YISFSGSTSYNPSLKS 424. PSMA-6 HCDR3 artificial aa WNYYGSSHVWFAY 425.
PSMA-6 LH artificial aa
DIKMTQSPSSMYASLGERVTITCKASQDIYSYLIWFQQKPGKSPKTLIYRANRLVDGVPSRFSGS
GSGQDYSLTISSLEYEDMGIYYCLQYDEFATFGSGTKLEMKGGGGSGGGGSGGGGSDVHLQESGP
GLVKPSQSLSLTCTVTGYSITSDYAWNWIRQFPGNKLEWMGYISFSGSTSYNPSLKSRISVTRDT
SKNQFFLQLNSVTTEDTATYYCARWNYYGSSHVWFAYWGQGTLVTVSS 426. PSMA-6 LH
artificial nt
GACATCAAGATGACCCAGTCCCCCTCCTCCATGTACGCCTCCCTGGGCGAGAGAGTGACCATCAC
CTGCAAGGCCTCCCAGGACATCTACTCCTACCTGATCTGGTTCCAGCAGAAGCCTGGCAAGTCCC
CTAAGACCCTGATCTACCGGGCCAACAGACTGGTGGACGGCGTGCCTTCCAGGTTCTCCGGCTCC
GGCTCTGGCCAGGACTACTCCCTGACCATCTCCTCCCTGGAATACGAGGACATGGGCATCTACTA
CTGCCTGCAGTACGACGAGTTCGCCACCTTCGGCTCCGGCACCAAGCTGGAAATGAAGGGCGGAG
GGGGATCTGGCGGCGGAGGAAGTGGCGGGGGAGGATCCGACGTGCACCTGCAGGAATCTGGCCCT
GGCCTGGTGAAGCCTTCCCAGTCCCTGTCCCTGACCTGCACCGTGACCGGCTACTCCATCACCTC
CGACTACGCCTGGAACTGGATCCGGCAGTTCCCTGGCAATAAGCTGGAATGGATGGGCTACATCT
CCTTCTCCGGCAGCACCTCCTACAACCCTTCCCTGAAGTCCCGGATCTCCGTGACCCGGGACACC
TCCAAGAACCAGTTCTTCCTGCAGCTGAACTCCGTGACCACCGAGGACACCGCCACCTACTACTG
CGCCCGGTGGAACTACTACGGCTCCTCCCACGTGTGGTTCGCTTACTGGGGCCAGGGCACCCTGG
TGACCGTGTCCTCC 427. PSMA-6 LH .times. I2C artificial aa
DIKMTQSPSSMYASLGERVTITCKASQDIYSYLIWFQQKPGKSPKTLIYRANRLVDGVPSRFSGS
HL
GSGQDYSLTISSLEYEDMGIYYCLQYDEFATFGSGTKLEMKGGGGSGGGGSGGGGSDVHLQESGP
GLVKPSQSLSLTCTVTGYSITSDYAWNWIRQFPGNKLEWMGYISFSGSTSYNPSLKSRISVTRDT
SKNQFFLQLNSVTTEDTATYYCARWNYYGSSHVWFAYWGQGTLVTVSSGGGGSEVQLVESGGGLV
QPGGSLKLSCAASGFTFNKYAMNWVRQAPGKGLEWVARIRSKYNNYATYYADSVKDRFTISRDDS
KNTAYLQMNNLKTEDTAVYYCVRHGNFGNSYISYWAYWGQGTLVTVSSGGGGSGGGGSGGGGSQT
VVTQEPSLTVSPGGTVTLTCGSSTGAVTSGNYPNWVQQKPGQAPRGLIGGTKFLAPGTPARFSGS
LLGGKAALTLSGVQPEDEAEYYCVLWYSNRWVFGGGTKLTVL 428. PSMA-6 LH .times.
I2C artificial nt
GACATCAAGATGACCCAGTCCCCCTCCTCCATGTACGCCTCCCTGGGCGAGAGAGTGACCATCAC
HL
CTGCAAGGCCTCCCAGGACATCTACTCCTACCTGATCTGGTTCCAGCAGAAGCCTGGCAAGTCCC
CTAAGACCCTGATCTACCGGGCCAACAGACTGGTGGACGGCGTGCCTTCCAGGTTCTCCGGCTCC
GGCTCTGGCCAGGACTACTCCCTGACCATCTCCTCCCTGGAATACGAGGACATGGGCATCTACTA
CTGCCTGCAGTACGACGAGTTCGCCACCTTCGGCTCCGGCACCAAGCTGGAAATGAAGGGCGGAG
GGGGATCTGGCGGCGGAGGAAGTGGCGGGGGAGGATCCGACGTGCACCTGCAGGAATCTGGCCCT
GGCCTGGTGAAGCCTTCCCAGTCCCTGTCCCTGACCTGCACCGTGACCGGCTACTCCATCACCTC
CGACTACGCCTGGAACTGGATCCGGCAGTTCCCTGGCAATAAGCTGGAATGGATGGGCTACATCT
CCTTCTCCGGCAGCACCTCCTACAACCCTTCCCTGAAGTCCCGGATCTCCGTGACCCGGGACACC
TCCAAGAACCAGTTCTTCCTGCAGCTGAACTCCGTGACCACCGAGGACACCGCCACCTACTACTG
CGCCCGGTGGAACTACTACGGCTCCTCCCACGTGTGGTTCGCTTACTGGGGCCAGGGCACCCTGC
TGACCGTGTCCTCCGGAGGTGGTGGATCCGAGGTGCAGCTGGTCGAGTCTGGAGGAGGATTGGTG
CAGCCTGGAGGGTCATTGAAACTCTCATGTGCAGCCTCTGGATTCACCTTCAATAAGTACGCCAT
GAACTGGGTCCGCCAGGCTCCAGGAAAGGGTTTGGAATGGGTTGCTCGCATAAGAAGTAAATATA
ATAATTATGCAACATATTATGCCGATTCAGTGAAAGACAGGTTCACCATCTCCAGAGATGATTCA
AAAAACACTGCCTATCTACAAATGAACAACTTGAAAACTGAGGACACTGCCGTGTACTACTGTGT
GAGACATGGGAACTTCGGTAATAGCTACATATCCTACTGGGCTTACTGGGGCCAAGGGACTCTGG
TCACCGTCTCCTCAGGTGGTGGTGGTTCTGGCGGCGGCGGCTCCGGTGGTGGTGGTTCTCAGACT
GTTGTGACTCAGGAACCTTCACTCACCGTATCACCTGGTGGAACAGTCACACTCACTTGTGGCTC
CTCGACTGGGGCTGTTACATCTGGCAACTACCCAAACTGGGTCCAACAAAAACCAGGTCAGGCAC
CCCGTGGTCTAATAGGTGGGACTAAGTTCCTCGCCCCCGGTACTCCTGCCAGATTCTCAGGCTCC
CTGCTTGGAGGCAAGGCTGCCCTCACCCTCTCAGGGGTACAGCCAGAGGATGAGGCAGAATATTA
CTGTGTTCTATGGTACAGCAACCGCTGGGTGTTCGGTGGAGGAACCAAACTGACTGTCCTA 429.
PSMA-7 L artificial aa
DIVMTQSHKFMSTSVGDRVSIICKASQDVGTAVDWYQQKPGQSPKLLIYWASTRHTGVPDRFTGS
GSGTDFTLTITNVQSEDLADYFCQQYNSYPLTFGAGTMLDLK 430. PSMA-7 L artificial
nt
GACATCGTGATGACCCAGTCCCACAAGTTCATGTCCACCTCCGTGGGCGACCGGGTGTCCATCAT
CTGCAAGGCCTCCCAGGATGTGGGCACCGCCGTGGACTGGTATCAGCAGAAGCCTGGCCAGTCCC
CTAAGCTGCTGATCTACTGGGCCTCCACCAGACACACCGGCGTGCCTGACAGATTCACCGGCTCC
GGCTCTGGCACCGACTTCACCCTGACCATCACCAACGTGCAGTCCGAGGACCTGGCCGACTACTT
CTGCCAGCAGTACAACTCCTACCCTCTGACCTTCGGCGCTGGCACCATGCTGGACCTGAAG 431.
PSMA-7 LCDR1 artificial aa KASQDVGTAVD 432. PSMA-7 LCDR2 artificial
aa WASTRHT 433. PSMA-7 LCDR3 artificial aa QQYNSYPLT 434. PSMA-7 H
artificial aa
EVQLQQSGPELVKPGTSVRISCKTSGYTFTEYTIHWVKQSHGKSLEWIGNINPNNGGTTYNQKFE
DKATLTVDKSSSTAYMELRSLTSEDSAVYYCAAGWNFDYWGQGTTLTVSS 435. PSMA-7 H
artificial nt
GAAGTGCAGCTGCAGCAGTCCGGCCCTGAGCTGGTGAAGCCTGGCACCTCCGTGCGGATCTCTTG
CAAGACCTCCGGCTACACCTTCACCGAGTACACCATCCACTGGGTGAAACAGTCCCACGGCAAGT
CCCTGGAATGGATCGGCAACATCAACCCTAACAACGGCGGCACCACCTACAACCAGAAGTTCGAG
GACAAGGCCACCCTGACCGTGGACAAGTCCTCCTCCACCGCCTACATGGAACTGCGGTCCCTGAC
CTCCGAGGACTCCGCCGTGTACTACTGCGCCGCTGGCTGGAACTTCGACTACTGGGGCCAGGGCA
CCACACTGACCGTGTCCTCC 436. PSMA-7 HCDR1 artificial aa EYTIH 437.
PSMA-7 HCDR2 artificial aa NINPNNGGTTYNQKFED 438. PSMA-7 HCDR3
artificial aa GWNFDY 439. PSMA-7 LH artificial aa
DIVMTQSHKFMSTSVGDRVSIICKASQDVGTAVDWYQQKPGQSPKLLIYWASTRHTGVPDRFTGS
GSGTDFTLTITNVQSEDLADYFCQQYNSYPLTFGAGTMLDLKGGGGSGGGGSGGGGSEVQLQQSG
PELVKPGTSVRISCKTSGYTFTEYTIHWVKQSHGKSLEWIGNINPNNGGTTYNQKFEDKATLTVD
KSSSTAYMELRSLTSEDSAVYYCAAGWNFDYWGQGTTLTVSS 440. PSMA-7 LH
artificial nt
GACATCGTGATGACCCAGTCCCACAAGTTCATGTCCACCTCCGTGGGCGACCGGGTGTCCATCAT
CTGCAAGGCCTCCCAGGATGTGGGCACCGCCGTGGACTGGTATCAGCAGAAGCCTGGCCAGTCCC
CTAAGCTGCTGATCTACTGGGCCTCCACCAGACACACCGGCGTGCCTGACAGATTCACCGGCTCC
GGCTCTGGCACCGACTTCACCCTGACCATCACCAACGTGCAGTCCGAGGACCTGGCCGACTACTT
CTGCCAGCAGTACAACTCCTACCCTCTGACCTTCGGCGCTGGCACCATGCTGGACCTGAAGGGCG
GAGGGGGCTCTGGCGGCGGAGGAAGTGGAGGGGGCGGATCCGAAGTGCAGCTGCAGCAGTCCGGC
CCTGAGCTGGTGAAGCCTGGCACCTCCGTGCGGATCTCTTGCAAGACCTCCGGCTACACCTTCAC
CGAGTACACCATCCACTGGGTGAAACAGTCCCACGGCAAGTCCCTGGAATGGATCGGCAACATCA
ACCCTAACAACGGCGGCACCACCTACAACCAGAAGTTCGAGGACAAGGCCACCCTGACCGTGGAC
AAGTCCTCCTCCACCGCCTACATGGAACTGCGGTCCCTGACCTCCGAGGACTCCGCCGTGTACTA
CTGCGCCGCTGGCTGGAACTTCGACTACTGGGGCCAGGGCACCACACTGACCGTGTCCTCC 441.
PSMA-7 LH .times. I2C artificial aa
DIVMTQSHKFMSTSVGDRVSIICKASQDVGTAVDWYQQKPGQSPKLLIYWASTRHTGVPDRFTGS
HL
GSGTDFTLTITNVQSEDLADYFCQQYNSYPLTFGAGTMLDLKGGGGSGGGGSGGGGSEVQLQQSG
PELVKPGTSVRISCKTSGYTFTEYTIHWVKQSHGKSLEWIGNINPNNGGTTYNQKFEDKATLTVD
KSSSTAYMELRSLTSEDSAVYYCAAGWNFDYWGQGTTLTVSSGGGGSEVQLVESGGGLVQPGGSL
KLSCAASGFTFNKYAMNWVRQAPGKGLEWVARIRSKYNNYATYYADSVKDRFTISRDDSKNTAYL
QMNNLKTEDTAVYYCVRHGNFGNSYISYWAYWGQGTLVTVSSGGGGSGGGGSGGGGSQTVVTQEP
SLTVSPGGTVTLTCGSSTGAVTSGNYPNWVQQKPGQAPRGLIGGTKFLAPGTPARFSGSLLGGKA
ALTLSGVQPEDEAEYYCVLWYSNRWVFGGGTKLTVL 442. PSMA-7 LH .times. I2C
artificial nt
GACATCGTGATGACCCAGTCCCACAAGTTCATGTCCACCTCCGTGGGCGACCGGGTGTCCATCAT
HL
CTGCAAGGCCTCCCAGGATGTGGGCACCGCCGTGGACTGGTATCAGCAGAAGCCTGGCCAGTCCC
CTAAGCTGCTGATCTACTGGGCCTCCACCAGACACACCGGCGTGCCTGACAGATTCACCGGCTCC
GGCTCTGGCACCGACTTCACCCTGACCATCACCAACGTGCAGTCCGAGGACCTGGCCGACTACTT
CTGCCAGCAGTACAACTCCTACCCTCTGACCTTCGGCGCTGGCACCATGCTGGACCTGAAGGGCG
GAGGGGGCTCTGGCGGCGGAGGAAGTGGAGGGGGCGGATCCGAAGTGCAGCTGCAGCAGTCCGGC
CCTGAGCTGGTGAAGCCTGGCACCTCCGTGCGGATCTCTTGCAAGACCTCCGGCTACACCTTCAC
CGAGTACACCATCCACTGGGTGAAACAGTCCCACGGCAAGTCCCTGGAATGGATCGGCAACATCA
ACCCTAACAACGGCGGCACCACCTACAACCAGAAGTTCGAGGACAAGGCCACCCTGACCGTGGAC
AAGTCCTCCTCCACCGCCTACATGGAACTGCGGTCCCTGACCTCCGAGGACTCCGCCGTGTACTA
CTGCGCCGCTGGCTGGAACTTCGACTACTGGGGCCAGGGCACCACACTGACCGTGTCCTCCGGAG
GTGGTGGATCCGAGGTGCAGCTGGTCGAGTCTGGAGGAGGATTGGTGCAGCCTGGAGGGTCATTG
AAACTCTCATGTGCAGCCTCTGGATTCACCTTCAATAAGTACGCCATGAACTGGGTCCGCCAGGC
TCCAGGAAAGGGTTTGGAATGGGTTGCTCGCATAAGAAGTAAATATAATAATTATGCAACATATT
ATGCCGATTCAGTGAAAGACAGGTTCACCATCTCCAGAGATGATTCAAAAAACACTGCCTATCTA
CAAATGAACAACTTGAAAACTGAGGACACTGCCGTGTACTACTGTGTGAGACATGGGAACTTCGG
TAATAGCTACATATCCTACTGGGCTTACTGGGGCCAAGGGACTCTGGTCACCGTCTCCTCAGGTG
GTGGTGGTTCTGGCGGCGGCGGCTCCGGTGGTGGTGGTTCTCAGACTGTTGTGACTCAGGAACCT
TCACTCACCGTATCACCTGGTGGAACAGTCACACTCACTTGTGGCTCCTCGACTGGGGCTGTTAC
ATCTGGCAACTACCCAAACTGGGTCCAACAAAAACCAGGTCAGGCACCCCGTGGTCTAATAGGTG
GGACTAAGTTCCTCGCCCCCGGTACTCCTGCCAGATTCTCAGGCTCCCTGCTTGGAGGCAAGGCT
GCCCTCACCCTCTCAGGGGTACAGCCAGAGGATGAGGCAGAATATTACTGTGTTCTATGGTACAG
CAACCGCTGGGTGTTCGGTGGAGGAACCAAACTGACTGTCCTA 443. PSMA-8 L
artificial aa
DIVLTQSPASLAVSLGQRATISCRASESIDSYDNTFMHWYQQKPGQPPNLLIFRASILESGIPAR
FSGSGSGTDFTLTIYPVEADDVATYYCHQSIEDPYTFGGGTKLEIK 444. PSMA-8 L
artificial nt
GACATCGTGCTGACCCAGTCTCCAGCCTCCCTGGCTGTGTCTCTGGGCCAGCGGGCCACCATCTC
TTGCCGGGCCTCCGAGTCCATCGACTCCTACGACAACACCTTCATGCACTGGTATCAGCAGAAGC
CTGGCCAGCCTCCTAACCTGCTGATCTTCCGGGCCTCTATCCTGGAATCCGGCATCCCTGCCCGG
TTCTCCGGCTCTGGCTCCGGCACCGACTTCACCCTGACCATCTACCCTGTGGAGGCCGACGACGT
GGCCACCTACTACTGCCACCAGTCCATCGAGGACCCTTACACCTTCGGCGGAGGGACCAAGCTGG
AAATCAAG 445. PSMA-8 LCDR1 artificial aa RASESIDSYDNTFMH 446.
PSMA-8 LCDR2 artificial aa RASILES 447. PSMA-8 LCDR3 artificial aa
HQSIEDPYT 448. PSMA-8 H artificial aa
EVQLQQSGPELVKPGASVKMSCKASGYTFTGYVMHWVKQKPGQVLEWIGYINPYNDVTRYNGKFK
GKATLTSDKYSSTAYMELSGLTSEDSAVYYCARGENWYYFDSWGRGATLTVSS 449. PSMA-8 H
artificial nt
GAAGTGCAGCTGCAGCAGTCCGGCCCTGAGCTGGTGAAGCCTGGCGCCTCCGTGAAGATGTCCTG
CAAGGCCTCCGGCTACACCTTCACCGGCTACGTGATGCACTGGGTGAAACAGAAACCCGGCCAGG
TGCTGGAATGGATCGGCTACATCAACCCTTACAACGACGTGACCCGGTACAACGGCAAGTTCAAG
GGCAAGGCCACCCTGACCTCCGACAAGTACTCCTCCACCGCCTACATGGAACTGTCCGGCCTGAC
CTCTGAGGACTCCGCCGTGTACTACTGCGCCAGGGGCGAGAACTGGTACTACTTCGACTCCTGGG
GCAGAGGCGCTACCCTGACCGTGTCTTCC 450. PSMA-8 HCDR1 artificial aa GYVMH
451. PSMA-8 HCDR2 artificial aa YINPYNDVTRYNGKFKG 452. PSMA-8 HCDR3
artificial aa GENWYYFDS 453. PSMA-8 LH artificial aa
DIVLTQSPASLAVSLGQRATISCRASESIDSYDNTFMHWYQQKPGQPPNLLIFRASILESGIPAR
FSGSGSGTDFTLTIYPVEADDVATYYCHQSIEDPYTFGGGTKLEIKGGGGSGGGGSGGGGSEVQL
QQSGPELVKPGASVKMSCKASGYTFTGYVMHWVKQKPGQVLEWIGYINPYNDVTRYNGKFKGKAT
LTSDKYSSTAYMELSGLTSEDSAVYYCARGENWYYFDSWGRGATLTVSS 454. PSMA-8 LH
artificial nt
GACATCGTGCTGACCCAGTCTCCAGCCTCCCTGGCTGTGTCTCTGGGCCAGCGGGCCACCATCTC
TTGCCGGGCCTCCGAGTCCATCGACTCCTACGACAACACCTTCATGCACTGGTATCAGCAGAAGC
CTGGCCAGCCTCCTAACCTGCTGATCTTCCGGGCCTCTATCCTGGAATCCGGCATCCCTGCCCGG
TTCTCCGGCTCTGGCTCCGGCACCGACTTCACCCTGACCATCTACCCTGTGGAGGCCGACGACGT
GGCCACCTACTACTGCCACCAGTCCATCGAGGACCCTTACACCTTCGGCGGAGGGACCAAGCTGG
AAATCAAGGGCGGAGGGGGATCTGGCGGCGGAGGAAGTGGAGGGGGCGGATCCGAAGTGCAGCTG
CAGCAGTCCGGCCCTGAGCTGGTGAAGCCTGGCGCCTCCGTGAAGATGTCCTGCAAGGCCTCCGG
CTACACCTTCACCGGCTACGTGATGCACTGGGTGAAACAGAAACCCGGCCAGGTGCTGGAATGGA
TCGGCTACATCAACCCTTACAACGACGTGACCCGGTACAACGGCAAGTTCAAGGGCAAGGCCACC
CTGACCTCCGACAAGTACTCCTCCACCGCCTACATGGAACTGTCCGGCCTGACCTCTGAGGACTC
CGCCGTGTACTACTGCGCCAGGGGCGAGAACTGGTACTACTTCGACTCCTGGGGCAGAGGCGCTA
CCCTGACCGTGTCTTCC 455. PSMA-8 LH .times. I2C artificial aa
DIVLTQSPASLAVSLGQRATISCRASESIDSYDNTFMHWYQQKPGQPPNLLIFRASILESGIPAR
HL
FSGSGSGTDFTLTIYPVEADDVATYYCHQSIEDPYTFGGGTKLEIKGGGGSGGGGSGGGGSEVQL
QQSGPELVKPGASVKMSCKASGYTFTGYVMHWVKQKPGQVLEWIGYINPYNDVTRYNGKFKGKAT
LTSDKYSSTAYMELSGLTSEDSAVYYCARGENWYYFDSWGRGATLTVSSGGGGSEVQLVESGGGL
VQPGGSLKLSCAASGFTFNKYAMNWVRQAPGKGLEWVARIRSKYNNYATYYADSVKDRFTISRDD
SKNTAYLQMNNLKTEDTAVYYCVRHGNFGNSYISYWAYWGQGTLVTVSSGGGGSGGGGSGGGGSQ
TVVTQEPSLTVSPGGTVTLTCGSSTGAVTSGNYPNWVQQKPGQAPRGLIGGTKFLAPGTPARFSG
SLLGGKAALTLSGVQPEDEAEYYCVLWYSNRWVFGGGTKLTVL 456. PSMA-8 LH .times.
I2C artificial nt
GACATCGTGCTGACCCAGTCTCCAGCCTCCCTGGCTGTGTCTCTGGGCCAGCGGGCCACCATCTC
HL
TTGCCGGGCCTCCGAGTCCATCGACTCCTACGACAACACCTTCATGCACTGGTATCAGCAGAAGC
CTGGCCAGCCTCCTAACCTGCTGATCTTCCGGGCCTCTATCCTGGAATCCGGCATCCCTGCCCGG
TTCTCCGGCTCTGGCTCCGGCACCGACTTCACCCTGACCATCTACCCTGTGGAGGCCGACGACGT
GGCCACCTACTACTGCCACCAGTCCATCGAGGACCCTTACACCTTCGGCGGAGGGACCAAGCTGG
AAATCAAGGGCGGAGGGGGATCTGGCGGCGGAGGAAGTGGAGGGGGCGGATCCGAAGTGCAGCTG
CAGCAGTCCGGCCCTGAGCTGGTGAAGCCTGGCGCCTCCGTGAAGATGTCCTGCAAGGCCTCCGG
CTACACCTTCACCGGCTACGTGATGCACTGGGTGAAACAGAAACCCGGCCAGGTGCTGGAATGGA
TCGGCTACATCAACCCTTACAACGACGTGACCCGGTACAACGGCAAGTTCAAGGGCAAGGCCACC
CTGACCTCCGACAAGTACTCCTCCACCGCCTACATGGAACTGTCCGGCCTGACCTCTGAGGACTC
CGCCGTGTACTACTGCGCCAGGGGCGAGAACTGGTACTACTTCGACTCCTGGGGCAGAGGCGCTA
CCCTGACCGTGTCTTCCGGAGGTGGTGGATCCGAGGTGCAGCTGGTCGAGTCTGGAGGAGGATTG
GTGCAGCCTGGAGGGTCATTGAAACTCTCATGTGCAGCCTCTGGATTCACCTTCAATAAGTACGC
CATGAACTGGGTCCGCCAGGCTCCAGGAAAGGGTTTGGAATGGGTTGCTCGCATAAGAAGTAAAT
ATAATAATTATGCAACATATTATGCCGATTCAGTGAAAGACAGGTTCACCATCTCCAGAGATGAT
TCAAAAAACACTGCCTATCTACAAATGAACAACTTGAAAACTGAGGACACTGCCGTGTACTACTG
TGTGAGACATGGGAACTTCGGTAATAGCTACATATCCTACTGGGCTTACTGGGGCCAAGGGACTC
TGGTCACCGTCTCCTCAGGTGGTGGTGGTTCTGGCGGCGGCGGCTCCGGTGGTGGTGGTTCTCAG
ACTGTTGTGACTCAGGAACCTTCACTCACCGTATCACCTGGTGGAACAGTCACACTCACTTGTGG
CTCCTCGACTGGGGCTGTTACATCTGGCAACTACCCAAACTGGGTCCAACAAAAACCAGGTCAGG
CACCCCGTGGTCTAATAGGTGGGACTAAGTTCCTCGCCCCCGGTACTCCTGCCAGATTCTCAGGC
TCCCTGCTTGGAGGCAAGGCTGCCCTCACCCTCTCAGGGGTACAGCCAGAGGATGAGGCAGAATA
TTACTGTGTTCTATGGTACAGCAACCGCTGGGTGTTCGGTGGAGGAACCAAACTGACTGTCCTA
457. PSMA-9 L artificial aa
NIVMTQSQKFMSTSPGDRVRVTCKASQNVGSDVAWYQAKPGQSPRILIYSTSYRYSGVPDRFTAY
GSGTDFTLTITNVQSEDLTEYFCQQYNSYPLTFGAGTKLELK 458. PSMA-9 L artificial
nt
AACATCGTGATGACCCAGTCCCAGAAATTCATGTCCACCTCTCCCGGCGATAGAGTGCGCGTGAC
CTGCAAGGCCTCCCAGAACGTGGGCTCCGACGTGGCCTGGTATCAGGCCAAGCCTGGCCAGTCCC
CTCGGATCCTGATCTACTCCACCTCCTACCGGTACTCCGGCGTGCCTGACAGATTCACCGCCTAC
GGCTCCGGCACCGACTTCACCCTGACCATTACAAACGTGCAGTCCGAGGACCTGACCGAGTACTT
CTGCCAGCAGTACAACTCCTACCCTCTGACCTTCGGCGCTGGCACCAAGCTGGAACTGAAG 459.
PSMA-9 LCDR1 artificial aa KASQNVGSDVA 460. PSMA-9 LCDR2 artificial
aa STSYRYS 461. PSMA-9 LCDR3 artificial aa QQYNSYPLT 462. PSMA-9 H
artificial aa
QVQLKESGPGLVASSQSLSITCTVSGFSLTAYGINWVRQPPGKGLEWLGVIWPDGNTDYNSTLKS
RLNIFKDNSKNQVFLKMSSFQTDDTARYFCARDSYGNFKRGWFDFWGQGTTLTVSS 463.
PSMA-9 H artificial nt
CAGGTGCAGCTGAAAGAGTCCGGCCCTGGCCTGGTGGCCTCCTCCCAGTCCCTGTCCATCACCTG
CACCGTGTCAGGCTTCTCCCTGACCGCTTACGGCATCAACTGGGTGCGCCAGCCTCCTGGCAAGG
GCCTGGAATGGCTGGGCGTGATCTGGCCTGACGGCAACACCGACTACAACAGCACCCTGAAGTCC
CGGCTGAACATCTTCAAGGACAACTCCAAGAACCAGGTGTTCCTGAAGATGTCCTCTTTCCAGAC
CGACGACACCGCCCGGTACTTTTGCGCCAGGGACTCCTACGGCAACTTCAAGCGGGGCTGGTTCG
ATTTTTGGGGCCAGGGCACCACACTGACCGTGTCCTCC 464. PSMA-9 HCDR1 artificial
aa AYGIN 465. PSMA-9 HCDR2 artificial aa VIWPDGNTDYNSTLKS 466.
PSMA-9 HCDR3 artificial aa DSYGNFKRGWFDF 467. PSMA-9 LH artificial
aa
NIVMTQSQKFMSTSPGDRVRVTCKASQNVGSDVAWYQAKPGQSPRILIYSTSYRYSGVPDRFTAY
GSGTDFTLTITNVQSEDLTEYFCQQYNSYPLTFGAGTKLELKGGGGSGGGGSGGGGSQVQLKESG
PGLVASSQSLSITCTVSGFSLTAYGINWVRQPPGKGLEWLGVIWPDGNTDYNSTLKSRLNIFKDN
SKNQVFLKMSSFQTDDTARYFCARDSYGNFKRGWFDFWGQGTTLTVSS 468. PSMA-9 LH
artificial nt
AACATCGTGATGACCCAGTCCCAGAAATTCATGTCCACCTCTCCCGGCGATAGAGTGCGCGTGAC
CTGCAAGGCCTCCCAGAACGTGGGCTCCGACGTGGCCTGGTATCAGGCCAAGCCTGGCCAGTCCC
CTCGGATCCTGATCTACTCCACCTCCTACCGGTACTCCGGCGTGCCTGACAGATTCACCGCCTAC
GGCTCCGGCACCGACTTCACCCTGACCATTACAAACGTGCAGTCCGAGGACCTGACCGAGTACTT
CTGCCAGCAGTACAACTCCTACCCTCTGACCTTCGGCGCTGGCACCAAGCTGGAACTGAAGGGCG
GAGGGGGCTCTGGCGGCGGAGGAAGTGGAGGGGGCGGATCTCAGGTGCAGCTGAAAGAGTCCGGC
CCTGGCCTGGTGGCCTCCTCCCAGTCCCTGTCCATCACCTGCACCGTGTCAGGCTTCTCCCTGAC
CGCTTACGGCATCAACTGGGTGCGCCAGCCTCCTGGCAAGGGCCTGGAATGGCTGGGCGTGATCT
GGCCTGACGGCAACACCGACTACAACAGCACCCTGAAGTCCCGGCTGAACATCTTCAAGGACAAC
TCCAAGAACCAGGTGTTCCTGAAGATGTCCTCTTTCCAGACCGACGACACCGCCCGGTACTTTTG
CGCCAGGGACTCCTACGGCAACTTCAAGCGGGGCTGGTTCGATTTTTGGGGCCAGGGCACCACAC
TGACCGTGTCCTCC 469. PSMA-9 LH .times. I2C artificial aa
NIVMTQSQKFMSTSPGDRVRVTCKASQNVGSDVAWYQAKPGQSPRILIYSTSYRYSGVPDRFTAY
HL
GSGTDFTLTITNVQSEDLTEYFCQQYNSYPLTFGAGTKLELKGGGGSGGGGSGGGGSQVQLKESG
PGLVASSQSLSITCTVSGFSLTAYGINWVRQPPGKGLEWLGVIWPDGNTDYNSTLKSRLNIFKDN
SKNQVFLKMSSFQTDDTARYFCARDSYGNFKRGWFDFWGQGTTLTVSSGGGGSEVQLVESGGGLV
QPGGSLKLSCAASGFTFNKYAMNWVRQAPGKGLEWVARIRSKYNNYATYYADSVKDRFTISRDDS
KNTAYLQMNNLKTEDTAVYYCVRHGNFGNSYISYWAYWGQGTLVTVSSGGGGSGGGGSGGGGSQT
VVTQEPSLTVSPGGTVTLTCGSSTGAVTSGNYPNWVQQKPGQAPRGLIGGTKFLAPGTPARFSGS
LLGGKAALTLSGVQPEDEAEYYCVLWYSNRWVFGGGTKLTVL 470. PSMA-9 LH .times.
I2C artificial nt
AACATCGTGATGACCCAGTCCCAGAAATTCATGTCCACCTCTCCCGGCGATAGAGTGCGCGTGAC
HL
CTGCAAGGCCTCCCAGAACGTGGGCTCCGACGTGGCCTGGTATCAGGCCAAGCCTGGCCAGTCCC
CTCGGATCCTGATCTACTCCACCTCCTACCGGTACTCCGGCGTGCCTGACAGATTCACCGCCTAC
GGCTCCGGCACCGACTTCACCCTGACCATTACAAACGTGCAGTCCGAGGACCTGACCGAGTACTT
CTGCCAGCAGTACAACTCCTACCCTCTGACCTTCGGCGCTGGCACCAAGCTGGAACTGAAGGGCG
GAGGGGGCTCTGGCGGCGGAGGAAGTGGAGGGGGCGGATCTCAGGTGCAGCTGAAAGAGTCCGGC
CCTGGCCTGGTGGCCTCCTCCCAGTCCCTGTCCATCACCTGCACCGTGTCAGGCTTCTCCCTGAC
CGCTTACGGCATCAACTGGGTGCGCCAGCCTCCTGGCAAGGGCCTGGAATGGCTGGGCGTGATCT
GGCCTGACGGCAACACCGACTACAACAGCACCCTGAAGTCCCGGCTGAACATCTTCAAGGACAAC
TCCAAGAACCAGGTGTTCCTGAAGATGTCCTCTTTCCAGACCGACGACACCGCCCGGTACTTTTG
CGCCAGGGACTCCTACGGCAACTTCAAGCGGGGCTGGTTCGATTTTTGGGGCCAGGGCACCACAC
TGACCGTGTCCTCCGGAGGTGGTGGATCCGAGGTGCAGCTGGTCGAGTCTGGAGGAGGATTGGTG
CAGCCTGGAGGGTCATTGAAACTCTCATGTGCAGCCTCTGGATTCACCTTCAATAAGTACGCCAT
GAACTGGGTCCGCCAGGCTCCAGGAAAGGGTTTGGAATGGGTTGCTCGCATAAGAAGTAAATATA
ATAATTATGCAACATATTATGCCGATTCAGTGAAAGACAGGTTCACCATCTCCAGAGATGATTCA
AAAAACACTGCCTATCTACAAATGAACAACTTGAAAACTGAGGACACTGCCGTGTACTACTGTGT
GAGACATGGGAACTTCGGTAATAGCTACATATCCTACTGGGCTTACTGGGGCCAAGGGACTCTGG
TCACCGTCTCCTCAGGTGGTGGTGGTTCTGGCGGCGGCGGCTCCGGTGGTGGTGGTTCTCAGACT
GTTGTGACTCAGGAACCTTCACTCACCGTATCACCTGGTGGAACAGTCACACTCACTTGTGGCTC
CTCGACTGGGGCTGTTACATCTGGCAACTACCCAAACTGGGTCCAACAAAAACCAGGTCAGGCAC
CCCGTGGTCTAATAGGTGGGACTAAGTTCCTCGCCCCCGGTACTCCTGCCAGATTCTCAGGCTCC
CTGCTTGGAGGCAAGGCTGCCCTCACCCTCTCAGGGGTACAGCCAGAGGATGAGGCAGAATATTA
CTGTGTTCTATGGTACAGCAACCGCTGGGTGTTCGGTGGAGGAACCAAACTGACTGTCCTA 471.
PSMA-10 VL artificial aa
NIVMTQFPKSMSISVGERVTLTCKASENVGTYVSWYQQKPEQSPKMLIYGASNRFTGVPDRFTGS
GSATDFILTISSVQTEDLVDYYCGQSYTFPYTFGGGTKLEMK 472. PSMA-10 VL
artificial nt
AACATCGTGATGACCCAGTTCCCTAAGTCCATGTCCATCTCCGTGGGCGAGAGAGTGACCCTGAC
CTGCAAGGCCTCCGAGAACGTGGGCACCTACGTGTCCTGGTATCAGCAGAAGCCTGAGCAGTCCC
CTAAGATGCTGATCTACGGCGCCTCCAACAGGTTCACCGGCGTGCCTGACAGATTCACCGGCTCC
GGCTCCGCCACCGACTTCATCCTGACCATCTCCAGCGTGCAGACCGAGGACCTGGTGGACTACTA
CTGCGGCCAGTCCTACACCTTCCCTTACACCTTCGGCGGAGGGACCAAGCTGGAAATGAAG 473.
PSMA-10 LCDR1 artificial aa KASENVGTYVS 474. PSMA-10 LCDR2
artificial aa GASNRFT 475. PSMA-10 LCDR3 artificial aa GQSYTFPYT
476. PSMA-10 VH artificial aa
EVKLEESGGGLVQPGGSMKLSCVASGFTFSNYWMNWVRQSPEKGLEWVAEIRSQSNNFATHYAES
VKGRVIISRDDSKSSVYLQMNNLRAEDTGIYYCTRRWNNFWGQGTTLTVSS 477. PSMA-10 VH
artificial nt
GAAGTGAAGCTGGAAGAGTCCGGCGGAGGACTGGTGCAGCCTGGCGGCTCCATGAAGCTGTCCTG
CGTGGCTTCCGGCTTCACCTTCTCCAACTACTGGATGAACTGGGTGCGCCAGTCCCCTGAGAAGG
GCCTGGAATGGGTGGCCGAGATCCGGTCCCAGTCCAACAACTTCGCCACCCACTACGCCGAGTCC
GTGAAGGGCAGAGTGATCATCTCCCGGGACGACTCCAAGTCCTCCGTGTACCTGCAGATGAACAA
CCTGCGGGCCGAGGACACCGGCATCTACTACTGCACCCGGCGGTGGAACAACTTTTGGGGCCAGG
GCACCACACTGACCGTGTCCTCC 478. PSMA-10 HCDR1 artificial aa NYWMN 479.
PSMA-10 HCDR2 artificial aa EIRSQSNNFATHYAESVKG 480. PSMA-10 HCDR3
artificial aa RWNNF 481. PSMA-10 LH artificial aa
NIVMTQFPKSMSISVGERVTLTCKASENVGTYVSWYQQKPEQSPKMLIYGASNRFTGVPDRFTGS
GSATDFILTISSVQTEDLVDYYCGQSYTFPYTFGGGTKLEMKGGGGSGGGGSGGGGSEVKLEESG
GGLVQPGGSMKLSCVASGFTFSNYWMNWVRQSPEKGLEWVAEIRSQSNNFATHYAESVKGRVIIS
RDDSKSSVYLQMNNLRAEDTGIYYCTRRWNNFWGQGTTLTVSS 482. PSMA-10 LH
artificial nt
AACATCGTGATGACCCAGTTCCCTAAGTCCATGTCCATCTCCGTGGGCGAGAGAGTGACCCTGAC
CTGCAAGGCCTCCGAGAACGTGGGCACCTACGTGTCCTGGTATCAGCAGAAGCCTGAGCAGTCCC
CTAAGATGCTGATCTACGGCGCCTCCAACAGGTTCACCGGCGTGCCTGACAGATTCACCGGCTCC
GGCTCCGCCACCGACTTCATCCTGACCATCTCCAGCGTGCAGACCGAGGACCTGGTGGACTACTA
CTGCGGCCAGTCCTACACCTTCCCTTACACCTTCGGCGGAGGGACCAAGCTGGAAATGAAGGGCG
GAGGGGGATCTGGCGGCGGAGGAAGTGGAGGGGGCGGATCCGAAGTGAAGCTGGAAGAGTCCGGC
GGAGGACTGGTGCAGCCTGGCGGCTCCATGAAGCTGTCCTGCGTGGCTTCCGGCTTCACCTTCTC
CAACTACTGGATGAACTGGGTGCGCCAGTCCCCTGAGAAGGGCCTGGAATGGGTGGCCGAGATCC
GGTCCCAGTCCAACAACTTCGCCACCCACTACGCCGAGTCCGTGAAGGGCAGAGTGATCATCTCC
CGGGACGACTCCAAGTCCTCCGTGTACCTGCAGATGAACAACCTGCGGGCCGAGGACACCGGCAT
CTACTACTGCACCCGGCGGTGGAACAACTTTTGGGGCCAGGGCACCACACTGACCGTGTCCTCC
483. PSMA-10 LH .times. I2C artificial aa
NIVMTQFPKSMSISVGERVTLTCKASENVGTYVSWYQQKPEQSPKMLIYGASNRFTGVPDRFTGS
HL
GSATDFILTISSVQTEDLVDYYCGQSYTFPYTFGGGTKLEMKGGGGSGGGGSGGGGSEVKLEESG
GGLVQPGGSMKLSCVASGFTFSNYWMNWVRQSPEKGLEWVAEIRSQSNNFATHYAESVKGRVIIS
RDDSKSSVYLQMNNLRAEDTGIYYCTRRWNNFWGQGTTLTVSSGGGGSEVQLVESGGGLVQPGGS
LKLSCAASGFTFNKYAMNWVRQAPGKGLEWVARIRSKYNNYATYYADSVKDRFTISRDDSKNTAY
LQMNNLKTEDTAVYYCVRHGNFGNSYISYWAYWGQGTLVTVSSGGGGSGGGGSGGGGSQTVVTQE
PSLTVSPGGTVTLTCGSSTGAVTSGNYPNWVQQKPGQAPRGLIGGTKFLAPGTPARFSGSLLGGK
AALTLSGVQPEDEAEYYCVLWYSNRWVFGGGTKLTVL 484. PSMA-10 LH .times. I2C
artificial nt
AACATCGTGATGACCCAGTTCCCTAAGTCCATGTCCATCTCCGTGGGCGAGAGAGTGACCCTGAC
HL
CTGCAAGGCCTCCGAGAACGTGGGCACCTACGTGTCCTGGTATCAGCAGAAGCCTGAGCAGTCCC
CTAAGATGCTGATCTACGGCGCCTCCAACAGGTTCACCGGCGTGCCTGACAGATTCACCGGCTCC
GGCTCCGCCACCGACTTCATCCTGACCATCTCCAGCGTGCAGACCGAGGACCTGGTGGACTACTA
CTGCGGCCAGTCCTACACCTTCCCTTACACCTTCGGCGGAGGGACCAAGCTGGAAATGAAGGGCG
GAGGGGGATCTGGCGGCGGAGGAAGTGGAGGGGGCGGATCCGAAGTGAAGCTGGAAGAGTCCGGC
GGAGGACTGGTGCAGCCTGGCGGCTCCATGAAGCTGTCCTGCGTGGCTTCCGGCTTCACCTTCTC
CAACTACTGGATGAACTGGGTGCGCCAGTCCCCTGAGAAGGGCCTGGAATGGGTGGCCGAGATCC
GGTCCCAGTCCAACAACTTCGCCACCCACTACGCCGAGTCCGTGAAGGGCAGAGTGATCATCTCC
CGGGACGACTCCAAGTCCTCCGTGTACCTGCAGATGAACAACCTGCGGGCCGAGGACACCGGCAT
CTACTACTGCACCCGGCGGTGGAACAACTTTTGGGGCCAGGGCACCACACTGACCGTGTCCTCCG
GAGGTGGTGGATCCGAGGTGCAGCTGGTCGAGTCTGGAGGAGGATTGGTGCAGCCTGGAGGGTCA
TTGAAACTCTCATGTGCAGCCTCTGGATTCACCTTCAATAAGTACGCCATGAACTGGGTCCGCCA
GGCTCCAGGAAAGGGTTTGGAATGGGTTGCTCGCATAAGAAGTAAATATAATAATTATGCAACAT
ATTATGCCGATTCAGTGAAAGACAGGTTCACCATCTCCAGAGATGATTCAAAAAACACTGCCTAT
CTACAAATGAACAACTTGAAAACTGAGGACACTGCCGTGTACTACTGTGTGAGACATGGGAACTT
CGGTAATAGCTACATATCCTACTGGGCTTACTGGGGCCAAGGGACTCTGGTCACCGTCTCCTCAG
GTGGTGGTGGTTCTGGCGGCGGCGGCTCCGGTGGTGGTGGTTCTCAGACTGTTGTGACTCAGGAA
CCTTCACTCACCGTATCACCTGGTGGAACAGTCACACTCACTTGTGGCTCCTCGACTGGGGCTGT
TACATCTGGCAACTACCCAAACTGGGTCCAACAAAAACCAGGTCAGGCACCCCGTGGTCTAATAG
GTGGGACTAAGTTCCTCGCCCCCGGTACTCCTGCCAGATTCTCAGGCTCCCTGCTTGGAGGCAAG
GCTGCCCTCACCCTCTCAGGGGTACAGCCAGAGGATGAGGCAGAATATTACTGTGTTCTATGGTA
CAGCAACCGCTGGGTGTTCGGTGGAGGAACCAAACTGACTGTCCTA 485. PSMA-A VL
artificial aa
DIQMTQSPSSLSASVGDRVTITCRASQGISNYLAWYQQKTGKVPKFLIYEASTLQSGVPSRFSGG
GSGTDFTLTISSLQPEDVATYYCQNYNSAPFTFGPGTKVDIK 486. PSMA-A VL
artificial nt
GACATCCAGATGACCCAGTCCCCCTCCTCCCTGTCTGCCTCCGTGGGCGACAGAGTGACCATCAC
CTGCCGGGCCTCCCAGGGCATCTCCAACTACCTGGCCTGGTATCAGCAGAAAACCGGCAAGGTGC
CCAAGTTCCTGATCTACGAGGCCTCCACCCTGCAGTCCGGCGTGCCTTCCAGATTCTCTGGCGGC
GGATCCGGCACCGACTTCACCCTGACCATCTCCAGCCTGCAGCCTGAGGACGTGGCCACCTACTA
CTGCCAGAACTACAACTCCGCCCCTTTCACCTTCGGCCCTGGCACCAAGGTGGACATCAAG 487.
PSMA-A LCDR1 artificial aa RASQGISNYLA 488. PSMA-A LCDR2 artificial
aa EASTLQS 489. PSMA-A LCDR3 artificial aa QNYNSAPFT 490. PSMA-A VH
artificial aa
QVQLVESGGGVVQPGRSLRLSCAASGFAFSRYGMHWVRQAPGKGLEWVAVIWYDGSNKYYADSVK
GRFTISRDNSKNTQYLQMNSLRAEDTAVYYCARGGDFLYYYYYGMDVWGQGTTVTVSS 491.
PSMA-A VH artificial nt
CAGGTGCAGCTGGTCGAGTCTGGCGGAGGGGTGGTCCAGCCTGGCCGGTCCCTGAGACTGTCTTG
CGCCGCCTCCGGCTTCGCCTTCTCCAGATACGGCATGCACTGGGTGCGCCAGGCTCCAGGCAAGG
GACTGGAATGGGTGGCCGTGATTTGGTACGACGGCTCCAACAAGTACTACGCCGACTCCGTGAAG
GGCCGGTTCACCATCTCCCGGGACAACTCCAAGAACACCCAGTACCTGCAGATGAACTCCCTGAG
GGCAGAGGACACCGCCGTGTACTACTGCGCCAGAGGCGGCGACTTCCTGTACTACTACTATTACG
GCATGGACGTGTGGGGCCAGGGCACCACCGTGACAGTGTCTTCC 492. PSMA-A HCDR1
artificial aa RYGMH 493. PSMA-A HCDR2 artificial aa
VIWYDGSNKYYADSVKG 494. PSMA-A HCDR3 artificial aa GGDFLYYYYYGMDV
495. PSMA-A LH artificial aa
DIQMTQSPSSLSASVGDRVTITCRASQGISNYLAWYQQKTGKVPKFLIYEASTLQSGVPSRFSGG
GSGTDFTLTISSLQPEDVATYYCQNYNSAPFTFGPGTKVDIKGGGGSGGGGSGGGGSQVQLVESG
GGVVQPGRSLRLSCAASGFAFSRYGMHWVRQAPGKGLEWVAVIWYDGSNKYYADSVKGRFTISRD
NSKNTQYLQMNSLRAEDTAVYYCARGGDFLYYYYYGMDVWGQGTTVTVSS 496. PSMA-A LH
artificial nt
GACATCCAGATGACCCAGTCCCCCTCCTCCCTGTCTGCCTCCGTGGGCGACAGAGTGACCATCAC
CTGCCGGGCCTCCCAGGGCATCTCCAACTACCTGGCCTGGTATCAGCAGAAAACCGGCAAGGTGC
CCAAGTTCCTGATCTACGAGGCCTCCACCCTGCAGTCCGGCGTGCCTTCCAGATTCTCTGGCGGC
GGATCCGGCACCGACTTCACCCTGACCATCTCCAGCCTGCAGCCTGAGGACGTGGCCACCTACTA
CTGCCAGAACTACAACTCCGCCCCTTTCACCTTCGGCCCTGGCACCAAGGTGGACATCAAGGGCG
GAGGGGGCAGTGGCGGCGGAGGAAGTGGAGGGGGCGGATCTCAGGTGCAGCTGGTCGAGTCTGGC
GGAGGGGTGGTCCAGCCTGGCCGGTCCCTGAGACTGTCTTGCGCCGCCTCCGGCTTCGCCTTCTC
CAGATACGGCATGCACTGGGTGCGCCAGGCTCCAGGCAAGGGACTGGAATGGGTGGCCGTGATTT
GGTACGACGGCTCCAACAAGTACTACGCCGACTCCGTGAAGGGCCGGTTCACCATCTCCCGGGAC
AACTCCAAGAACACCCAGTACCTGCAGATGAACTCCCTGAGGGCAGAGGACACCGCCGTGTACTA
CTGCGCCAGAGGCGGCGACTTCCTGTACTACTACTATTACGGCATGGACGTGTGGGGCCAGGGCA
CCACCGTGACAGTGTCTTCC 497. PSMA-A LH .times. I2C artificial aa
DIQMTQSPSSLSASVGDRVTITCRASQGISNYLAWYQQKTGKVPKFLIYEASTLQSGVPSRFSGG
HL
GSGTDFTLTISSLQPEDVATYYCQNYNSAPFTFGPGTKVDIKGGGGSGGGGSGGGGSQVQLVESG
GGVVQPGRSLRLSCAASGFAFSRYGMHWVRQAPGKGLEWVAVIWYDGSNKYYADSVKGRFTISRD
NSKNTQYLQMNSLRAEDTAVYYCARGGDFLYYYYYGMDVWGQGTTVTVSSGGGGSEVQLVESGGG
LVQPGGSLKLSCAASGFTFNKYAMNWVRQAPGKGLEWVARIRSKYNNYATYYADSVKDRFTISRD
DSKNTAYLQMNNLKTEDTAVYYCVRHGNFGNSYISYWAYWGQGTLVTVSSGGGGSGGGGSGGGGS
QTVVTQEPSLTVSPGGTVTLTCGSSTGAVTSGNYPNWVQQKPGQAPRGLIGGTKFLAPGTPARFS
GSLLGGKAALTLSGVQPEDEAEYYCVLWYSNRWVFGGGTKLTVL 498. PSMA-A LH .times.
I2C artificial nt
GACATCCAGATGACCCAGTCCCCCTCCTCCCTGTCTGCCTCCGTGGGCGACAGAGTGACCATCAC
HL
CTGCCGGGCCTCCCAGGGCATCTCCAACTACCTGGCCTGGTATCAGCAGAAAACCGGCAAGGTGC
CCAAGTTCCTGATCTACGAGGCCTCCACCCTGCAGTCCGGCGTGCCTTCCAGATTCTCTGGCGGC
GGATCCGGCACCGACTTCACCCTGACCATCTCCAGCCTGCAGCCTGAGGACGTGGCCACCTACTA
CTGCCAGAACTACAACTCCGCCCCTTTCACCTTCGGCCCTGGCACCAAGGTGGACATCAAGGGCG
GAGGGGGCAGTGGCGGCGGAGGAAGTGGAGGGGGCGGATCTCAGGTGCAGCTGGTCGAGTCTGGC
GGAGGGGTGGTCCAGCCTGGCCGGTCCCTGAGACTGTCTTGCGCCGCCTCCGGCTTCGCCTTCTC
CAGATACGGCATGCACTGGGTGCGCCAGGCTCCAGGCAAGGGACTGGAATGGGTGGCCGTGATTT
GGTACGACGGCTCCAACAAGTACTACGCCGACTCCGTGAAGGGCCGGTTCACCATCTCCCGGGAC
AACTCCAAGAACACCCAGTACCTGCAGATGAACTCCCTGAGGGCAGAGGACACCGCCGTGTACTA
CTGCGCCAGAGGCGGCGACTTCCTGTACTACTACTATTACGGCATGGACGTGTGGGGCCAGGGCA
CCACCGTGACAGTGTCTTCCGGAGGTGGTGGATCCGAGGTGCAGCTGGTCGAGTCTGGAGGAGGA
TTGGTGCAGCCTGGAGGGTCATTGAAACTCTCATGTGCAGCCTCTGGATTCACCTTCAATAAGTA
CGCCATGAACTGGGTCCGCCAGGCTCCAGGAAAGGGTTTGGAATGGGTTGCTCGCATAAGAAGTA
AATATAATAATTATGCAACATATTATGCCGATTCAGTGAAAGACAGGTTCACCATCTCCAGAGAT
GATTCAAAAAACACTGCCTATCTACAAATGAACAACTTGAAAACTGAGGACACTGCCGTGTACTA
CTGTGTGAGACATGGGAACTTCGGTAATAGCTACATATCCTACTGGGCTTACTGGGGCCAAGGGA
CTCTGGTCACCGTCTCCTCAGGTGGTGGTGGTTCTGGCGGCGGCGGCTCCGGTGGTGGTGGTTCT
CAGACTGTTGTGACTCAGGAACCTTCACTCACCGTATCACCTGGTGGAACAGTCACACTCACTTG
TGGCTCCTCGACTGGGGCTGTTACATCTGGCAACTACCCAAACTGGGTCCAACAAAAACCAGGTC
AGGCACCCCGTGGTCTAATAGGTGGGACTAAGTTCCTCGCCCCCGGTACTCCTGCCAGATTCTCA
GGCTCCCTGCTTGGAGGCAAGGCTGCCCTCACCCTCTCAGGGGTACAGCCAGAGGATGAGGCAGA
ATATTACTGTGTTCTATGGTACAGCAACCGCTGGGTGTTCGGTGGAGGAACCAAACTGACTGTCC
TA 499. PSMA-B VL artificial aa
DIQMTQSPSSLSASVGDRVTITCRASQGITNYLAWFQQKPGKAPKSLIYAASSLQSGVPSKFSGS
GSGTDFSLTISSLQPEDFATYYCQQYNSYPITFGQGTRLEIK 500. PSMA-B VL
artificial nt
GACATCCAGATGACCCAGTCACCCTCCTCCCTGTCTGCCTCCGTGGGCGACAGAGTGACCATCAC
CTGCCGGGCCTCACAGGGCATCACCAACTACCTGGCCTGGTTCCAGCAGAAGCCTGGCAAGGCCC
CTAAGTCCCTGATCTACGCCGCCTCCTCTCTGCAGTCCGGCGTGCCTTCCAAGTTCTCCGGCTCC
GGCTCTGGCACCGACTTCTCCCTGACCATCTCCTCCCTGCAGCCTGAGGACTTCGCCACCTACTA
CTGCCAGCAGTACAACTCCTACCCTATCACCTTCGGCCAGGGCACCCGGCTGGAAATCAAG 501.
PSMA-B LCDR1 artificial aa RASQGITNYLA 502. PSMA-B LCDR2 artificial
aa AASSLQS 503. PSMA-B LCDR3 artificial aa QQYNSYPIT 504. PSMA-B VH
artificial aa
QVQLVESGGGVVQPGRSLRLSCAASGFTFSNYVMHWVRQAPGKGLEWVAIIWYDGSNKYYADSVK
GRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAGGYNWNYEYHYYGMDVWGQGTTVTVSS 505.
PSMA-B VH artificial nt
CAGGTGCAGCTGGTCGAGTCTGGCGGAGGGGTGGTGCAGCCTGGCCGGTCCCTGAGACTGTCTTG
CGCTGCCTCCGGCTTCACCTTCTCCAACTACGTGATGCACTGGGTGCGCCAGGCTCCAGGCAAGG
GACTGGAATGGGTGGCCATCATTTGGTACGACGGCTCCAACAAGTACTACGCCGACTCCGTGAAG
GGCCGGTTCACCATCTCCCGGGACAACTCCAAGAACACCCTGTACCTGCAGATGAACTCCCTGAG
GGCCGAGGACACCGCCGTGTACTACTGCGCTGGCGGCTACAACTGGAACTACGAGTACCACTACT
ACGGCATGGACGTGTGGGGCCAGGGAACCACCGTGACCGTGTCTTCC 506. PSMA-B HCDR1
artificial aa NYVMH 507. PSMA-B HCDR2 artificial aa
IIWYDGSNKYYADSVKG 508. PSMA-B HCDR3 artificial aa GYNWNYEYHYYGMDV
509. PSMA-B LH artificial aa
DIQMTQSPSSLSASVGDRVTITCRASQGITNYLAWFQQKPGKAPKSLIYAASSLQSGVPSKFSGS
GSGTDFSLTISSLQPEDFATYYCQQYNSYPITFGQGTRLEIKGGGGSGGGGSGGGGSQVQLVESG
GGVVQPGRSLRLSCAASGFTFSNYVMHWVRQAPGKGLEWVAIIWYDGSNKYYADSVKGRFTISRD
NSKNTLYLQMNSLRAEDTAVYYCAGGYNWNYEYHYYGMDVWGQGTTVTVSS 510. PSMA-B LH
artificial nt
GACATCCAGATGACCCAGTCACCCTCCTCCCTGTCTGCCTCCGTGGGCGACAGAGTGACCATCAC
CTGCCGGGCCTCACAGGGCATCACCAACTACCTGGCCTGGTTCCAGCAGAAGCCTGGCAAGGCCC
CTAAGTCCCTGATCTACGCCGCCTCCTCTCTGCAGTCCGGCGTGCCTTCCAAGTTCTCCGGCTCC
GGCTCTGGCACCGACTTCTCCCTGACCATCTCCTCCCTGCAGCCTGAGGACTTCGCCACCTACTA
CTGCCAGCAGTACAACTCCTACCCTATCACCTTCGGCCAGGGCACCCGGCTGGAAATCAAGGGCG
GAGGGGGATCTGGCGGCGGAGGAAGTGGAGGGGGCGGATCTCAGGTGCAGCTGGTCGAGTCTGGC
GGAGGGGTGGTGCAGCCTGGCCGGTCCCTGAGACTGTCTTGCGCTGCCTCCGGCTTCACCTTCTC
CAACTACGTGATGCACTGGGTGCGCCAGGCTCCAGGCAAGGGACTGGAATGGGTGGCCATCATTT
GGTACGACGGCTCCAACAAGTACTACGCCGACTCCGTGAAGGGCCGGTTCACCATCTCCCGGGAC
AACTCCAAGAACACCCTGTACCTGCAGATGAACTCCCTGAGGGCCGAGGACACCGCCGTGTACTA
CTGCGCTGGCGGCTACAACTGGAACTACGAGTACCACTACTACGGCATGGACGTGTGGGGCCAGG
GAACCACCGTGACCGTGTCTTCC 511. PSMA-B LH .times. I2C artificial aa
DIQMTQSPSSLSASVGDRVTITCRASQGITNYLAWFQQKPGKAPKSLIYAASSLQSGVPSKFSGS
HL
GSGTDFSLTISSLQPEDFATYYCQQYNSYPITFGQGTRLEIKGGGGSGGGGSGGGGSQVQLVESG
GGVVQPGRSLRLSCAASGFTFSNYVMHWVRQAPGKGLEWVAIIWYDGSNKYYADSVKGRFTISRD
NSKNTLYLQMNSLRAEDTAVYYCAGGYNWNYEYHYYGMDVWGQGTTVTVSSGGGGSEVQLVESGG
GLVQPGGSLKLSCAASGFTFNKYAMNWVRQAPGKGLEWVARIRSKYNNYATYYADSVKDRFTISR
DDSKNTAYLQMNNLKTEDTAVYYCVRHGNFGNSYISYWAYWGQGTLVTVSSGGGGSGGGGSGGGG
SQTVVTQEPSLTVSPGGTVTLTCGSSTGAVTSGNYPNWVQQKPGQAPRGLIGGTKFLAPGTPARF
SGSLLGGKAALTLSGVQPEDEAEYYCVLWYSNRWVFGGGTKLTVL 512. PSMA-B LH
.times. I2C artificial nt
GACATCCAGATGACCCAGTCACCCTCCTCCCTGTCTGCCTCCGTGGGCGACAGAGTGACCATCAC
HL
CTGCCGGGCCTCACAGGGCATCACCAACTACCTGGCCTGGTTCCAGCAGAAGCCTGGCAAGGCCC
CTAAGTCCCTGATCTACGCCGCCTCCTCTCTGCAGTCCGGCGTGCCTTCCAAGTTCTCCGGCTCC
GGCTCTGGCACCGACTTCTCCCTGACCATCTCCTCCCTGCAGCCTGAGGACTTCGCCACCTACTA
CTGCCAGCAGTACAACTCCTACCCTATCACCTTCGGCCAGGGCACCCGGCTGGAAATCAAGGGCG
GAGGGGGATCTGGCGGCGGAGGAAGTGGAGGGGGCGGATCTCAGGTGCAGCTGGTCGAGTCTGGC
GGAGGGGTGGTGCAGCCTGGCCGGTCCCTGAGACTGTCTTGCGCTGCCTCCGGCTTCACCTTCTC
CAACTACGTGATGCACTGGGTGCGCCAGGCTCCAGGCAAGGGACTGGAATGGGTGGCCATCATTT
GGTACGACGGCTCCAACAAGTACTACGCCGACTCCGTGAAGGGCCGGTTCACCATCTCCCGGGAC
AACTCCAAGAACACCCTGTACCTGCAGATGAACTCCCTGAGGGCCGAGGACACCGCCGTGTACTA
CTGCGCTGGCGGCTACAACTGGAACTACGAGTACCACTACTACGGCATGGACGTGTGGGGCCAGG
GAACCACCGTGACCGTGTCTTCCGGAGGTGGTGGATCCGAGGTGCAGCTGGTCGAGTCTGGAGGA
GGATTGGTGCAGCCTGGAGGGTCATTGAAACTCTCATGTGCAGCCTCTGGATTCACCTTCAATAA
GTACGCCATGAACTGGGTCCGCCAGGCTCCAGGAAAGGGTTTGGAATGGGTTGCTCGCATAAGAA
GTAAATATAATAATTATGCAACATATTATGCCGATTCAGTGAAAGACAGGTTCACCATCTCCAGA
GATGATTCAAAAAACACTGCCTATCTACAAATGAACAACTTGAAAACTGAGGACACTGCCGTGTA
CTACTGTGTGAGACATGGGAACTTCGGTAATAGCTACATATCCTACTGGGCTTACTGGGGCCAAG
GGACTCTGGTCACCGTCTCCTCAGGTGGTGGTGGTTCTGGCGGCGGCGGCTCCGGTGGTGGTGGT
TCTCAGACTGTTGTGACTCAGGAACCTTCACTCACCGTATCACCTGGTGGAACAGTCACACTCAC
TTGTGGCTCCTCGACTGGGGCTGTTACATCTGGCAACTACCCAAACTGGGTCCAACAAAAACCAG
GTCAGGCACCCCGTGGTCTAATAGGTGGGACTAAGTTCCTCGCCCCCGGTACTCCTGCCAGATTC
TCAGGCTCCCTGCTTGGAGGCAAGGCTGCCCTCACCCTCTCAGGGGTACAGCCAGAGGATGAGGC
AGAATATTACTGTGTTCTATGGTACAGCAACCGCTGGGTGTTCGGTGGAGGAACCAAACTGACTG
TCCTA 513. PSMA-C VL artificial aa
DIQMTQSPSSLSASVGDRVTITCRASQGISHYLAWFQQKPGKAPKSLIYAASSLQSGVPSKFSGS
GSGTDFTLTISSLQPEDFATYYCQQYNSFPLTFGGGTKVEIK 514. PSMA-C VL
artificial nt
GACATCCAGATGACCCAGTCTCCCTCCTCCCTGTCTGCCTCCGTGGGCGACAGAGTGACCATCAC
CTGCCGGGCCTCCCAGGGCATCTCTCACTACCTGGCCTGGTTCCAGCAGAAGCCTGGCAAGGCCC
CTAAGTCCCTGATCTACGCCGCCTCCTCTCTGCAGTCCGGCGTGCCTTCCAAGTTCTCCGGCTCC
GGCTCTGGCACCGACTTCACCCTGACCATCTCCAGCCTGCAGCCTGAGGACTTCGCCACCTACTA
CTGCCAGCAGTACAACTCCTTCCCTCTGACCTTCGGCGGAGGGACCAAGGTGGAGATCAAG 515.
PSMA-C LCDR1 artificial aa RASQGISHYLA 516. PSMA-C LCDR2 artificial
aa AASSLQS 517. PSMA-C LCDR3 artificial aa QQYNSFPLT 518. PSMA-C VH
artificial aa
QVQLVESGGGVVQPGRSLRLSCAASGFTFSSYGMHWVRQAPGKGLDWVAIIWHDGSNKYYADSVK
GRFTISRDNSKKTLYLQMNSLRAEDTAVYYCARAWAYDYGDYEYYFGMDVWGQGTTVTVSS 519.
PSMA-C VH artificial nt
CAGGTGCAGCTGGTCGAGTCTGGCGGAGGGGTGGTGCAGCCTGGCCGGTCCCTGAGACTGTCTTG
TGCCGCCTCCGGCTTCACCTTCTCCTCTTACGGCATGCACTGGGTGCGCCAGGCTCCAGGCAAGG
GACTGGACTGGGTGGCCATCATCTGGCACGACGGCTCCAACAAGTACTACGCCGACTCCGTGAAG
GGCCGGTTCACCATCTCCCGGGACAACTCCAAGAAAACCCTGTACCTGCAGATGAACTCCCTGAG
GGCCGAGGACACCGCCGTGTACTACTGTGCCAGGGCCTGGGCCTACGACTACGGCGACTACGAGT
ACTACTTCGGCATGGACGTGTGGGGCCAGGGCACCACCGTGACAGTGTCTTCC 520. PSMA-C
HCDR1 artificial aa SYGMH 521. PSMA-C HCDR2 artificial aa
IIWHDGSNKYYADSVKG 522. PSMA-C HCDR3 artificial aa AWAYDYGDYEYYFGMDV
523. PSMA-C LH artificial aa
DIQMTQSPSSLSASVGDRVTITCRASQGISHYLAWFQQKPGKAPKSLIYAASSLQSGVPSKFSGS
GSGTDFTLTISSLQPEDFATYYCQQYNSFPLTFGGGTKVEIKGGGGSGGGGSGGGGSQVQLVESG
GGVVQPGRSLRLSCAASGFTFSSYGMHWVRQAPGKGLDWVAIIWHDGSNKYYADSVKGRFTISRD
NSKKTLYLQMNSLRAEDTAVYYCARAWAYDYGDYEYYFGMDVWGQGTTVTVSS 524. PSMA-C
LH artificial nt
GACATCCAGATGACCCAGTCTCCCTCCTCCCTGTCTGCCTCCGTGGGCGACAGAGTGACCATCAC
CTGCCGGGCCTCCCAGGGCATCTCTCACTACCTGGCCTGGTTCCAGCAGAAGCCTGGCAAGGCCC
CTAAGTCCCTGATCTACGCCGCCTCCTCTCTGCAGTCCGGCGTGCCTTCCAAGTTCTCCGGCTCC
GGCTCTGGCACCGACTTCACCCTGACCATCTCCAGCCTGCAGCCTGAGGACTTCGCCACCTACTA
CTGCCAGCAGTACAACTCCTTCCCTCTGACCTTCGGCGGAGGGACCAAGGTGGAGATCAAGGGCG
GAGGGGGCTCTGGCGGCGGAGGAAGTGGAGGGGGCGGATCTCAGGTGCAGCTGGTCGAGTCTGGC
GGAGGGGTGGTGCAGCCTGGCCGGTCCCTGAGACTGTCTTGTGCCGCCTCCGGCTTCACCTTCTC
CTCTTACGGCATGCACTGGGTGCGCCAGGCTCCAGGCAAGGGACTGGACTGGGTGGCCATCATCT
GGCACGACGGCTCCAACAAGTACTACGCCGACTCCGTGAAGGGCCGGTTCACCATCTCCCGGGAC
AACTCCAAGAAAACCCTGTACCTGCAGATGAACTCCCTGAGGGCCGAGGACACCGCCGTGTACTA
CTGTGCCAGGGCCTGGGCCTACGACTACGGCGACTACGAGTACTACTTCGGCATGGACGTGTGGG
GCCAGGGCACCACCGTGACAGTGTCTTCC 525. PSMA-C LH .times. I2C artificial
aa
DIQMTQSPSSLSASVGDRVTITCRASQGISHYLAWFQQKPGKAPKSLIYAASSLQSGVPSKFSGS
HL
GSGTDFTLTISSLQPEDFATYYCQQYNSFPLTFGGGTKVEIKGGGGSGGGGSGGGGSQVQLVESG
GGVVQPGRSLRLSCAASGFTFSSYGMHWVRQAPGKGLDWVAIIWHDGSNKYYADSVKGRFTISRD
NSKKTLYLQMNSLRAEDTAVYYCARAWAYDYGDYEYYFGMDVWGQGTTVTVSSGGGGSEVQLVES
GGGLVQPGGSLKLSCAASGFTFNKYAMNWVRQAPGKGLEWVARIRSKYNNYATYYADSVKDRFTI
SRDDSKNTAYLQMNNLKTEDTAVYYCVRHGNFGNSYISYWAYWGQGTLVTVSSGGGGSGGGGSGG
GGSQTVVTQEPSLTVSPGGTVTLTCGSSTGAVTSGNYPNWVQQKPGQAPRGLIGGTKFLAPGTPA
RFSGSLLGGKAALTLSGVQPEDEAEYYCVLWYSNRWVFGGGTKLTVL 526. PSMA-C LH
.times. I2C artificial nt
GACATCCAGATGACCCAGTCTCCCTCCTCCCTGTCTGCCTCCGTGGGCGACAGAGTGACCATCAC
HL
CTGCCGGGCCTCCCAGGGCATCTCTCACTACCTGGCCTGGTTCCAGCAGAAGCCTGGCAAGGCCC
CTAAGTCCCTGATCTACGCCGCCTCCTCTCTGCAGTCCGGCGTGCCTTCCAAGTTCTCCGGCTCC
GGCTCTGGCACCGACTTCACCCTGACCATCTCCAGCCTGCAGCCTGAGGACTTCGCCACCTACTA
CTGCCAGCAGTACAACTCCTTCCCTCTGACCTTCGGCGGAGGGACCAAGGTGGAGATCAAGGGCG
GAGGGGGCTCTGGCGGCGGAGGAAGTGGAGGGGGCGGATCTCAGGTGCAGCTGGTCGAGTCTGGC
GGAGGGGTGGTGCAGCCTGGCCGGTCCCTGAGACTGTCTTGTGCCGCCTCCGGCTTCACCTTCTC
CTCTTACGGCATGCACTGGGTGCGCCAGGCTCCAGGCAAGGGACTGGACTGGGTGGCCATCATCT
GGCACGACGGCTCCAACAAGTACTACGCCGACTCCGTGAAGGGCCGGTTCACCATCTCCCGGGAC
AACTCCAAGAAAACCCTGTACCTGCAGATGAACTCCCTGAGGGCCGAGGACACCGCCGTGTACTA
CTGTGCCAGGGCCTGGGCCTACGACTACGGCGACTACGAGTACTACTTCGGCATGGACGTGTGGG
GCCAGGGCACCACCGTGACAGTGTCTTCCGGAGGTGGTGGATCCGAGGTGCAGCTGGTCGAGTCT
GGAGGAGGATTGGTGCAGCCTGGAGGGTCATTGAAACTCTCATGTGCAGCCTCTGGATTCACCTT
CAATAAGTACGCCATGAACTGGGTCCGCCAGGCTCCAGGAAAGGGTTTGGAATGGGTTGCTCGCA
TAAGAAGTAAATATAATAATTATGCAACATATTATGCCGATTCAGTGAAAGACAGGTTCACCATC
TCCAGAGATGATTCAAAAAACACTGCCTATCTACAAATGAACAACTTGAAAACTGAGGACACTGC
CGTGTACTACTGTGTGAGACATGGGAACTTCGGTAATAGCTACATATCCTACTGGGCTTACTGGG
GCCAAGGGACTCTGGTCACCGTCTCCTCAGGTGGTGGTGGTTCTGGCGGCGGCGGCTCCGGTGGT
GGTGGTTCTCAGACTGTTGTGACTCAGGAACCTTCACTCACCGTATCACCTGGTGGAACAGTCAC
ACTCACTTGTGGCTCCTCGACTGGGGCTGTTACATCTGGCAACTACCCAAACTGGGTCCAACAAA
AACCAGGTCAGGCACCCCGTGGTCTAATAGGTGGGACTAAGTTCCTCGCCCCCGGTACTCCTGCC
AGATTCTCAGGCTCCCTGCTTGGAGGCAAGGCTGCCCTCACCCTCTCAGGGGTACAGCCAGAGGA
TGAGGCAGAATATTACTGTGTTCTATGGTACAGCAACCGCTGGGTGTTCGGTGGAGGAACCAAAC
TGACTGTCCTA 527. PSMA-D VL artificial aa
GIVMTQSPATLSVSPGERATLSCRTSQSIGWNLAWYQQKPGQAPRLLIYGASSRTTGIPARFSGS
GSGTEFTLTISSLQSEDSAVYYCQHYDNWPMCSFGQGTELEIK 528. PSMA-D VL
artificial nt
GGCATCGTGATGACCCAGTCCCCCGCCACCCTGTCTGTGTCTCCCGGCGAGAGAGCCACCCTGAG
CTGCCGGACCTCCCAGTCCATCGGCTGGAACCTGGCCTGGTATCAGCAGAAGCCTGGACAGGCCC
CTAGACTGCTGATCTACGGCGCCTCCTCCAGAACCACCGGCATCCCTGCCAGGTTCTCCGGCTCT
GGCTCCGGCACCGAGTTCACCCTGACCATCTCCAGCCTGCAGTCCGAGGACTCCGCCGTGTACTA
CTGCCAGCACTACGACAACTGGCCTATGTGCTCCTTCGGCCAGGGCACCGAGCTGGAAATCAAG
529. PSMA-D LCDR1 artificial aa RTSQSIGWNLA 530. PSMA-D LCDR2
artificial aa GASSRTT 531. PSMA-D LCDR3 artificial aa QHYDNWPMCS
532. PSMA-D VH artificial aa
EVQLVQSGAEVKKPGESLKISCKGSGYSFTSYWIGWVRQMPGKGLEWMGIIYPGDSDTRYSPSFQ
GQVTISADKSISTAYLQWSSLKASDTAMYYCARRMAAAGPFDYWGQGTLVTVSS 533. PSMA-D
VH artificial nt
GAAGTGCAGCTGGTGCAGTCTGGCGCCGAAGTGAAGAAGCCCGGCGAGTCCCTGAAGATCTCCTG
CAAGGGCTCCGGCTACTCCTTCACCTCCTACTGGATCGGCTGGGTGCGCCAGATGCCTGGCAAGG
GCCTGGAATGGATGGGCATCATCTACCCTGGCGACTCCGACACCCGGTACTCTCCCAGCTTCCAG
GGCCAGGTGACCATCTCTGCCGACAAGTCCATCTCCACCGCCTACCTGCAGTGGTCCTCCCTGAA
GGCCTCCGACACCGCCATGTACTATTGCGCCAGGCGGATGGCCGCTGCCGGCCCTTTTGATTACT
GGGGCCAGGGAACCCTGGTGACCGTGTCCTCC 534. PSMA-D HCDR1 artificial aa
SYWIG 535. PSMA-D HCDR2 artificial aa IIYPGDSDTRYSPSFQG 536. PSMA-D
HCDR3 artificial aa RMAAAGPFDY 537. PSMA-D LH artificial aa
GIVMTQSPATLSVSPGERATLSCRTSQSIGWNLAWYQQKPGQAPRLLIYGASSRTTGIPARFSGS
GSGTEFTLTISSLQSEDSAVYYCQHYDNWPMCSFGQGTELEIKGGGGSGGGGSGGGGSEVQLVQS
GAEVKKPGESLKISCKGSGYSFTSYWIGWVRQMPGKGLEWMGIIYPGDSDTRYSPSFQGQVTISA
DKSISTAYLQWSSLKASDTAMYYCARRMAAAGPFDYWGQGTLVTVSS 538. PSMA-D LH
artificial nt
GGCATCGTGATGACCCAGTCCCCCGCCACCCTGTCTGTGTCTCCCGGCGAGAGAGCCACCCTGAG
CTGCCGGACCTCCCAGTCCATCGGCTGGAACCTGGCCTGGTATCAGCAGAAGCCTGGACAGGCCC
CTAGACTGCTGATCTACGGCGCCTCCTCCAGAACCACCGGCATCCCTGCCAGGTTCTCCGGCTCT
GGCTCCGGCACCGAGTTCACCCTGACCATCTCCAGCCTGCAGTCCGAGGACTCCGCCGTGTACTA
CTGCCAGCACTACGACAACTGGCCTATGTGCTCCTTCGGCCAGGGCACCGAGCTGGAAATCAAGG
GCGGAGGGGGATCTGGCGGCGGAGGAAGTGGAGGGGGCGGATCCGAAGTGCAGCTGGTGCAGTCT
GGCGCCGAAGTGAAGAAGCCCGGCGAGTCCCTGAAGATCTCCTGCAAGGGCTCCGGCTACTCCTT
CACCTCCTACTGGATCGGCTGGGTGCGCCAGATGCCTGGCAAGGGCCTGGAATGGATGGGCATCA
TCTACCCTGGCGACTCCGACACCCGGTACTCTCCCAGCTTCCAGGGCCAGGTGACCATCTCTGCC
GACAAGTCCATCTCCACCGCCTACCTGCAGTGGTCCTCCCTGAAGGCCTCCGACACCGCCATGTA
CTATTGCGCCAGGCGGATGGCCGCTGCCGGCCCTTTTGATTACTGGGGCCAGGGAACCCTGGTGA
CCGTGTCCTCC 539. PSMA-D LH .times. I2C artificial aa
GIVMTQSPATLSVSPGERATLSCRTSQSIGWNLAWYQQKPGQAPRLLIYGASSRTTGIPARFSGS
HL
GSGTEFTLTISSLQSEDSAVYYCQHYDNWPMCSFGQGTELEIKGGGGSGGGGSGGGGSEVQLVQS
GAEVKKPGESLKISCKGSGYSFTSYWIGWVRQMPGKGLEWMGIIYPGDSDTRYSPSFQGQVTISA
DKSISTAYLQWSSLKASDTAMYYCARRMAAAGPFDYWGQGTLVTVSSGGGGSEVQLVESGGGLVQ
PGGSLKLSCAASGFTFNKYAMNWVRQAPGKGLEWVARIRSKYNNYATYYADSVKDRFTISRDDSK
NTAYLQMNNLKTEDTAVYYCVRHGNFGNSYISYWAYWGQGTLVTVSSGGGGSGGGGSGGGGSQTV
VTQEPSLTVSPGGTVTLTCGSSTGAVTSGNYPNWVQQKPGQAPRGLIGGTKFLAPGTPARFSGSL
LGGKAALTLSGVQPEDEAEYYCVLWYSNRWVFGGGTKLTVL 540. PSMA-D LH .times.
I2C artificial nt
GGCATCGTGATGACCCAGTCCCCCGCCACCCTGTCTGTGTCTCCCGGCGAGAGAGCCACCCTGAG
HL
CTGCCGGACCTCCCAGTCCATCGGCTGGAACCTGGCCTGGTATCAGCAGAAGCCTGGACAGGCCC
CTAGACTGCTGATCTACGGCGCCTCCTCCAGAACCACCGGCATCCCTGCCAGGTTCTCCGGCTCT
GGCTCCGGCACCGAGTTCACCCTGACCATCTCCAGCCTGCAGTCCGAGGACTCCGCCGTGTACTA
CTGCCAGCACTACGACAACTGGCCTATGTGCTCCTTCGGCCAGGGCACCGAGCTGGAAATCAAGG
GCGGAGGGGGATCTGGCGGCGGAGGAAGTGGAGGGGGCGGATCCGAAGTGCAGCTGGTGCAGTCT
GGCGCCGAAGTGAAGAAGCCCGGCGAGTCCCTGAAGATCTCCTGCAAGGGCTCCGGCTACTCCTT
CACCTCCTACTGGATCGGCTGGGTGCGCCAGATGCCTGGCAAGGGCCTGGAATGGATGGGCATCA
TCTACCCTGGCGACTCCGACACCCGGTACTCTCCCAGCTTCCAGGGCCAGGTGACCATCTCTGCC
GACAAGTCCATCTCCACCGCCTACCTGCAGTGGTCCTCCCTGAAGGCCTCCGACACCGCCATGTA
CTATTGCGCCAGGCGGATGGCCGCTGCCGGCCCTTTTGATTACTGGGGCCAGGGAACCCTGGTGA
CCGTGTCCTCCGGAGGTGGTGGATCCGAGGTGCAGCTGGTCGAGTCTGGAGGAGGATTGGTGCAG
CCTGGAGGGTCATTGAAACTCTCATGTGCAGCCTCTGGATTCACCTTCAATAAGTACGCCATGAA
CTGGGTCCGCCAGGCTCCAGGAAAGGGTTTGGAATGGGTTGCTCGCATAAGAAGTAAATATAATA
ATTATGCAACATATTATGCCGATTCAGTGAAAGACAGGTTCACCATCTCCAGAGATGATTCAAAA
AACACTGCCTATCTACAAATGAACAACTTGAAAACTGAGGACACTGCCGTGTACTACTGTGTGAG
ACATGGGAACTTCGGTAATAGCTACATATCCTACTGGGCTTACTGGGGCCAAGGGACTCTGGTCA
CCGTCTCCTCAGGTGGTGGTGGTTCTGGCGGCGGCGGCTCCGGTGGTGGTGGTTCTCAGACTGTT
GTGACTCAGGAACCTTCACTCACCGTATCACCTGGTGGAACAGTCACACTCACTTGTGGCTCCTC
GACTGGGGCTGTTACATCTGGCAACTACCCAAACTGGGTCCAACAAAAACCAGGTCAGGCACCCC
GTGGTCTAATAGGTGGGACTAAGTTCCTCGCCCCCGGTACTCCTGCCAGATTCTCAGGCTCCCTG
CTTGGAGGCAAGGCTGCCCTCACCCTCTCAGGGGTACAGCCAGAGGATGAGGCAGAATATTACTG
TGTTCTATGGTACAGCAACCGCTGGGTGTTCGGTGGAGGAACCAAACTGACTGTCCTA 541.
PSMA-E VL artificial aa
DIVMTQTPLSLSVTPGQPASISCKSSQSLLHSDGKTFLYWYLQKPGQPPQLLIYEVSNRFSGVPD
RFSGSGSGTDFTLKISRVEAEDVGLYYCMQSIQLPLTFGGGTKVEIK 542. PSMA-E VL
artificial nt
GACATCGTGATGACCCAGACCCCTCTGTCCCTGTCTGTGACCCCTGGCCAGCCTGCCTCCATCTC
CTGCAAGTCCTCCCAGTCCCTGCTGCACTCCGACGGCAAGACCTTCCTGTACTGGTATCTGCAGA
AGCCCGGCCAGCCTCCTCAGCTGCTGATCTACGAGGTGTCCAACCGGTTCTCCGGCGTGCCTGAC
AGGTTCTCTGGCTCCGGCTCCGGCACCGACTTCACCCTGAAGATCTCCCGGGTGGAGGCTGAGGA
CGTGGGCCTGTACTACTGCATGCAGTCCATCCAGCTGCCTCTGACCTTCGGCGGAGGGACCAAGG
TGGAGATCAAG 543. PSMA-E LCDR1 artificial aa KSSQSLLHSDGKTFLY 544.
PSMA-E LCDR2 artificial aa EVSNRFS 545. PSMA-E LCDR3 artificial aa
MQSIQLPLT 546. PSMA-E VH artificial aa
QVQLVESGGGVVQPGRSLRLSCAASGFTFISYGMHWVRQAPGKGLEWVAVISYDGSNKYYADSVK
GRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARVLVGALYYYNYYGMDVWGQGTTVTVSS 547.
PSMA-E VH artificial nt
CAGGTGCAGCTGGTCGAGTCTGGGGGAGGGGTCGTGCAGCCTGGCCGGTCCCTGAGACTGTCTTG
CGCCGCCTCCGGCTTCACCTTCATCTCTTACGGCATGCACTGGGTGCGCCAGGCTCCAGGCAAGG
GACTGGAATGGGTGGCCGTGATCTCCTACGACGGCTCCAACAAGTACTACGCCGACTCCGTGAAG
GGCCGGTTCACCATCTCCCGGGACAACTCCAAGAACACCCTGTACCTGCAGATGAACTCCCTGAG
GGCCGAGGACACCGCCGTGTACTACTGTGCCAGGGTGCTGGTCGGCGCTCTGTACTACTACAACT
ACTACGGCATGGACGTGTGGGGCCAGGGCACCACCGTGACAGTGTCTTCC 548. PSMA-E
HCDR1 artificial aa SYGMH 549. PSMA-E HCDR2 artificial aa
VISYDGSNKYYADSVKG 550. PSMA-E HCDR3 artificial aa VLVGALYYYNYYGMDV
551. PSMA-E LH artificial aa
DIVMTQTPLSLSVTPGQPASISCKSSQSLLHSDGKTFLYWYLQKPGQPPQLLIYEVSNRFSGVPD
RFSGSGSGTDFTLKISRVEAEDVGLYYCMQSIQLPLTFGGGTKVEIKGGGGSGGGGSGGGGSQVQ
LVESGGGVVQPGRSLRLSCAASGFTFISYGMHWVRQAPGKGLEWVAVISYDGSNKYYADSVKGRF
TISRDNSKNTLYLQMNSLRAEDTAVYYCARVLVGALYYYNYYGMDVWGQGTTVTVSS 552.
PSMA-E LH artificial nt
GACATCGTGATGACCCAGACCCCTCTGTCCCTGTCTGTGACCCCTGGCCAGCCTGCCTCCATCTC
CTGCAAGTCCTCCCAGTCCCTGCTGCACTCCGACGGCAAGACCTTCCTGTACTGGTATCTGCAGA
AGCCCGGCCAGCCTCCTCAGCTGCTGATCTACGAGGTGTCCAACCGGTTCTCCGGCGTGCCTGAC
AGGTTCTCTGGCTCCGGCTCCGGCACCGACTTCACCCTGAAGATCTCCCGGGTGGAGGCTGAGGA
CGTGGGCCTGTACTACTGCATGCAGTCCATCCAGCTGCCTCTGACCTTCGGCGGAGGGACCAAGG
TGGAGATCAAGGGCGGAGGGGGCTCTGGCGGCGGAGGAAGTGGAGGGGGCGGATCTCAGGTGCAG
CTGGTCGAGTCTGGGGGAGGGGTCGTGCAGCCTGGCCGGTCCCTGAGACTGTCTTGCGCCGCCTC
CGGCTTCACCTTCATCTCTTACGGCATGCACTGGGTGCGCCAGGCTCCAGGCAAGGGACTGGAAT
GGGTGGCCGTGATCTCCTACGACGGCTCCAACAAGTACTACGCCGACTCCGTGAAGGGCCGGTTC
ACCATCTCCCGGGACAACTCCAAGAACACCCTGTACCTGCAGATGAACTCCCTGAGGGCCGAGGA
CACCGCCGTGTACTACTGTGCCAGGGTGCTGGTCGGCGCTCTGTACTACTACAACTACTACGGCA
TGGACGTGTGGGGCCAGGGCACCACCGTGACAGTGTCTTCC 553. PSMA-E LH .times.
I2C artificial aa
DIVMTQTPLSLSVTPGQPASISCKSSQSLLHSDGKTFLYWYLQKPGQPPQLLIYEVSNRFSGVPD
HL
RFSGSGSGTDFTLKISRVEAEDVGLYYCMQSIQLPLTFGGGTKVEIKGGGGSGGGGSGGGGSQVQ
LVESGGGVVQPGRSLRLSCAASGFTFISYGMHWVRQAPGKGLEWVAVISYDGSNKYYADSVKGRF
TISRDNSKNTLYLQMNSLRAEDTAVYYCARVLVGALYYYNYYGMDVWGQGTTVTVSSGGGGSEVQ
LVESGGGLVQPGGSLKLSCAASGFTFNKYAMNWVRQAPGKGLEWVARIRSKYNNYATYYADSVKD
RFTISRDDSKNTAYLQMNNLKTEDTAVYYCVRHGNFGNSYISYWAYWGQGTLVTVSSGGGGSGGG
GSGGGGSQTVVTQEPSLTVSPGGTVTLTCGSSTGAVTSGNYPNWVQQKPGQAPRGLIGGTKFLAP
GTPARFSGSLLGGKAALTLSGVQPEDEAEYYCVLWYSNRWVFGGGTKLTVL 554. PSMA-E LH
.times. I2C artificial nt
GACATCGTGATGACCCAGACCCCTCTGTCCCTGTCTGTGACCCCTGGCCAGCCTGCCTCCATCTC
HL
CTGCAAGTCCTCCCAGTCCCTGCTGCACTCCGACGGCAAGACCTTCCTGTACTGGTATCTGCAGA
AGCCCGGCCAGCCTCCTCAGCTGCTGATCTACGAGGTGTCCAACCGGTTCTCCGGCGTGCCTGAC
AGGTTCTCTGGCTCCGGCTCCGGCACCGACTTCACCCTGAAGATCTCCCGGGTGGAGGCTGAGGA
CGTGGGCCTGTACTACTGCATGCAGTCCATCCAGCTGCCTCTGACCTTCGGCGGAGGGACCAAGG
TGGAGATCAAGGGCGGAGGGGGCTCTGGCGGCGGAGGAAGTGGAGGGGGCGGATCTCAGGTGCAG
CTGGTCGAGTCTGGGGGAGGGGTCGTGCAGCCTGGCCGGTCCCTGAGACTGTCTTGCGCCGCCTC
CGGCTTCACCTTCATCTCTTACGGCATGCACTGGGTGCGCCAGGCTCCAGGCAAGGGACTGGAAT
GGGTGGCCGTGATCTCCTACGACGGCTCCAACAAGTACTACGCCGACTCCGTGAAGGGCCGGTTC
ACCATCTCCCGGGACAACTCCAAGAACACCCTGTACCTGCAGATGAACTCCCTGAGGGCCGAGGA
CACCGCCGTGTACTACTGTGCCAGGGTGCTGGTCGGCGCTCTGTACTACTACAACTACTACGGCA
TGGACGTGTGGGGCCAGGGCACCACCGTGACAGTGTCTTCCGGAGGTGGTGGATCCGAGGTGCAG
CTGGTCGAGTCTGGAGGAGGATTGGTGCAGCCTGGAGGGTCATTGAAACTCTCATGTGCAGCCTC
TGGATTCACCTTCAATAAGTACGCCATGAACTGGGTCCGCCAGGCTCCAGGAAAGGGTTTGGAAT
GGGTTGCTCGCATAAGAAGTAAATATAATAATTATGCAACATATTATGCCGATTCAGTGAAAGAC
AGGTTCACCATCTCCAGAGATGATTCAAAAAACACTGCCTATCTACAAATGAACAACTTGAAAAC
TGAGGACACTGCCGTGTACTACTGTGTGAGACATGGGAACTTCGGTAATAGCTACATATCCTACT
GGGCTTACTGGGGCCAAGGGACTCTGGTCACCGTCTCCTCAGGTGGTGGTGGTTCTGGCGGCGGC
GGCTCCGGTGGTGGTGGTTCTCAGACTGTTGTGACTCAGGAACCTTCACTCACCGTATCACCTGG
TGGAACAGTCACACTCACTTGTGGCTCCTCGACTGGGGCTGTTACATCTGGCAACTACCCAAACT
GGGTCCAACAAAAACCAGGTCAGGCACCCCGTGGTCTAATAGGTGGGACTAAGTTCCTCGCCCCC
GGTACTCCTGCCAGATTCTCAGGCTCCCTGCTTGGAGGCAAGGCTGCCCTCACCCTCTCAGGGGT
ACAGCCAGAGGATGAGGCAGAATATTACTGTGTTCTATGGTACAGCAACCGCTGGGTGTTCGGTG
GAGGAACCAAACTGACTGTCCTA 555. PSMA-F VL artificial aa
AIQLTQSPSSLSASVGDRVTITCRASQGISSALAWYQQKSGKAPKLLIFDASSLESGVPSRFSGS
GSGTDFTLTISSLQPEDFATYYCQQFNSYPLTFGGGTKVEIK 556. PSMA-F VL
artificial nt
GCCATCCAGCTGACCCAGAGTCCCTCCTCCCTGTCTGCCTCCGTGGGCGACAGAGTGACCATCAC
CTGCCGGGCCTCCCAGGGCATCTCTTCCGCCCTGGCCTGGTATCAGCAGAAGTCCGGCAAGGCCC
CTAAGCTGCTGATCTTCGACGCCTCCTCTCTGGAATCCGGCGTGCCTTCCCGGTTCTCCGGCTCT
GGCTCCGGCACCGACTTCACCCTGACCATCTCCAGCCTGCAGCCTGAGGACTTCGCCACCTACTA
CTGCCAGCAGTTCAACTCCTACCCTCTGACCTTCGGCGGAGGGACCAAGGTGGAGATCAAG 557.
PSMA-F LCDR1 artificial aa RASQGISSALA 558. PSMA-F LCDR2 artificial
aa DASSLES 559. PSMA-F LCDR3 artificial aa QQFNSYPLT 560. PSMA-F VH
artificial aa
QVQLVESGGGVVQPGRSLRLSCAASGFTFSSYAMHWVRQAPGKGLEWVAVISYDGNNKYYADSVK
GRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARAVPWGSRYYYYGMDVWGQGTTVTVSS 561.
PSMA-F VH artificial nt
CAGGTGCAGCTGGTCGAGTCTGGGGGAGGGGTCGTGCAGCCTGGCCGGTCCCTGAGACTGTCTTG
CGCCGCCTCCGGCTTCACCTTCTCCTCTTACGCCATGCACTGGGTGCGCCAGGCTCCAGGCAAGG
GACTGGAATGGGTGGCCGTGATCTCCTACGACGGCAACAACAAGTACTACGCCGACTCCGTGAAG
GGCCGGTTCACCATCTCCCGGGACAACTCCAAGAACACCCTGTACCTGCAGATGAACTCCCTGAG
GGCTGAGGACACCGCCGTGTACTACTGCGCCAGAGCCGTGCCTTGGGGCTCCCGGTACTACTACT
ACGGCATGGACGTGTGGGGCCAGGGCACCACCGTGACAGTGTCTTCC 562. PSMA-F HCDR1
artificial aa SYAMH 563. PSMA-F HCDR2 artificial aa
VISYDGNNKYYADSVKG 564. PSMA-F HCDR3 artificial aa AVPWGSRYYYYGMDV
565. PSMA-F LH artificial aa
AIQLTQSPSSLSASVGDRVTITCRASQGISSALAWYQQKSGKAPKLLIFDASSLESGVPSRFSGS
GSGTDFTLTISSLQPEDFATYYCQQFNSYPLTFGGGTKVEIKGGGGSGGGGSGGGGSQVQLVESG
GGVVQPGRSLRLSCAASGFTFSSYAMHWVRQAPGKGLEWVAVISYDGNNKYYADSVKGRFTISRD
NSKNTLYLQMNSLRAEDTAVYYCARAVPWGSRYYYYGMDVWGQGTTVTVSS 566. PSMA-F LH
artificial nt
GCCATCCAGCTGACCCAGAGTCCCTCCTCCCTGTCTGCCTCCGTGGGCGACAGAGTGACCATCAC
CTGCCGGGCCTCCCAGGGCATCTCTTCCGCCCTGGCCTGGTATCAGCAGAAGTCCGGCAAGGCCC
CTAAGCTGCTGATCTTCGACGCCTCCTCTCTGGAATCCGGCGTGCCTTCCCGGTTCTCCGGCTCT
GGCTCCGGCACCGACTTCACCCTGACCATCTCCAGCCTGCAGCCTGAGGACTTCGCCACCTACTA
CTGCCAGCAGTTCAACTCCTACCCTCTGACCTTCGGCGGAGGGACCAAGGTGGAGATCAAGGGCG
GAGGGGGCTCTGGCGGCGGAGGAAGTGGAGGGGGCGGATCTCAGGTGCAGCTGGTCGAGTCTGGG
GGAGGGGTCGTGCAGCCTGGCCGGTCCCTGAGACTGTCTTGCGCCGCCTCCGGCTTCACCTTCTC
CTCTTACGCCATGCACTGGGTGCGCCAGGCTCCAGGCAAGGGACTGGAATGGGTGGCCGTGATCT
CCTACGACGGCAACAACAAGTACTACGCCGACTCCGTGAAGGGCCGGTTCACCATCTCCCGGGAC
AACTCCAAGAACACCCTGTACCTGCAGATGAACTCCCTGAGGGCTGAGGACACCGCCGTGTACTA
CTGCGCCAGAGCCGTGCCTTGGGGCTCCCGGTACTACTACTACGGCATGGACGTGTGGGGCCAGG
GCACCACCGTGACAGTGTCTTCC 567. PSMA-F LH .times. I2C artificial aa
AIQLTQSPSSLSASVGDRVTITCRASQGISSALAWYQQKSGKAPKLLIFDASSLESGVPSRFSGS
HL
GSGTDFTLTISSLQPEDFATYYCQQFNSYPLTFGGGTKVEIKGGGGSGGGGSGGGGSQVQLVESG
GGVVQPGRSLRLSCAASGFTFSSYAMHWVRQAPGKGLEWVAVISYDGNNKYYADSVKGRFTISRD
NSKNTLYLQMNSLRAEDTAVYYCARAVPWGSRYYYYGMDVWGQGTTVTVSSGGGGSEVQLVESGG
GLVQPGGSLKLSCAASGFTFNKYAMNWVRQAPGKGLEWVARIRSKYNNYATYYADSVKDRFTISR
DDSKNTAYLQMNNLKTEDTAVYYCVRHGNFGNSYISYWAYWGQGTLVTVSSGGGGSGGGGSGGGG
SQTVVTQEPSLTVSPGGTVTLTCGSSTGAVTSGNYPNWVQQKPGQAPRGLIGGTKFLAPGTPARF
SGSLLGGKAALTLSGVQPEDEAEYYCVLWYSNRWVFGGGTKLTVL 568. PSMA-F LH
.times. I2C artificial nt
GCCATCCAGCTGACCCAGAGTCCCTCCTCCCTGTCTGCCTCCGTGGGCGACAGAGTGACCATCAC
HL
CTGCCGGGCCTCCCAGGGCATCTCTTCCGCCCTGGCCTGGTATCAGCAGAAGTCCGGCAAGGCCC
CTAAGCTGCTGATCTTCGACGCCTCCTCTCTGGAATCCGGCGTGCCTTCCCGGTTCTCCGGCTCT
GGCTCCGGCACCGACTTCACCCTGACCATCTCCAGCCTGCAGCCTGAGGACTTCGCCACCTACTA
CTGCCAGCAGTTCAACTCCTACCCTCTGACCTTCGGCGGAGGGACCAAGGTGGAGATCAAGGGCG
GAGGGGGCTCTGGCGGCGGAGGAAGTGGAGGGGGCGGATCTCAGGTGCAGCTGGTCGAGTCTGGG
GGAGGGGTCGTGCAGCCTGGCCGGTCCCTGAGACTGTCTTGCGCCGCCTCCGGCTTCACCTTCTC
CTCTTACGCCATGCACTGGGTGCGCCAGGCTCCAGGCAAGGGACTGGAATGGGTGGCCGTGATCT
CCTACGACGGCAACAACAAGTACTACGCCGACTCCGTGAAGGGCCGGTTCACCATCTCCCGGGAC
AACTCCAAGAACACCCTGTACCTGCAGATGAACTCCCTGAGGGCTGAGGACACCGCCGTGTACTA
CTGCGCCAGAGCCGTGCCTTGGGGCTCCCGGTACTACTACTACGGCATGGACGTGTGGGGCCAGG
GCACCACCGTGACAGTGTCTTCCGGAGGTGGTGGATCCGAGGTGCAGCTGGTCGAGTCTGGAGGA
GGATTGGTGCAGCCTGGAGGGTCATTGAAACTCTCATGTGCAGCCTCTGGATTCACCTTCAATAA
GTACGCCATGAACTGGGTCCGCCAGGCTCCAGGAAAGGGTTTGGAATGGGTTGCTCGCATAAGAA
GTAAATATAATAATTATGCAACATATTATGCCGATTCAGTGAAAGACAGGTTCACCATCTCCAGA
GATGATTCAAAAAACACTGCCTATCTACAAATGAACAACTTGAAAACTGAGGACACTGCCGTGTA
CTACTGTGTGAGACATGGGAACTTCGGTAATAGCTACATATCCTACTGGGCTTACTGGGGCCAAG
GGACTCTGGTCACCGTCTCCTCAGGTGGTGGTGGTTCTGGCGGCGGCGGCTCCGGTGGTGGTGGT
TCTCAGACTGTTGTGACTCAGGAACCTTCACTCACCGTATCACCTGGTGGAACAGTCACACTCAC
TTGTGGCTCCTCGACTGGGGCTGTTACATCTGGCAACTACCCAAACTGGGTCCAACAAAAACCAG
GTCAGGCACCCCGTGGTCTAATAGGTGGGACTAAGTTCCTCGCCCCCGGTACTCCTGCCAGATTC
TCAGGCTCCCTGCTTGGAGGCAAGGCTGCCCTCACCCTCTCAGGGGTACAGCCAGAGGATGAGGC
AGAATATTACTGTGTTCTATGGTACAGCAACCGCTGGGTGTTCGGTGGAGGAACCAAACTGACTG
TCCTA 569. PSMA-J VL artificial aa
EIVLTQSPATLSLSPGERATLSCRASQSVSSYLAWFQQKPGQAPRLLIYDASNRATGIPARFSGS
GSGTDFTLTISSLEPEDFAVYYCQQRSNWLMYTFGQGTKLEIK 570. PSMA-J VL
artificial nt
GAGATCGTGCTGACCCAGTCCCCTGCCACCCTGTCTCTGTCTCCCGGCGAGAGAGCCACCCTGAG
CTGCCGGGCCTCCCAGTCCGTGTCCTCCTACCTGGCCTGGTTCCAGCAGAAGCCTGGACAGGCCC
CTAGGCTGCTGATCTACGACGCCTCCAACAGGGCTACCGGCATCCCTGCCCGGTTCTCCGGCTCT
GGCTCCGGCACCGACTTCACCCTGACCATCTCCAGCCTGGAACCTGAGGACTTCGCCGTGTACTA
CTGCCAGCAGCGGTCCAACTGGCTGATGTATACCTTCGGCCAGGGCACCAAGCTGGAAATCAAG
571. PSMA-J LCDR1 artificial aa RASQSVSSYLA 572. PSMA-J LCDR2
artificial aa DASNRAT
573. PSMA-J LCDR3 artificial aa QQRSNWLMYT 574. PSMA-J VH
artificial aa
EVQLVQSGAEVKKPGESLKISCKGSGYSFTSYWIGWARQMPGKGLEWMGIIYPGDSDTRYSPSFQ
GQVTISADKSISTAYLQWSSLKASDTAMYYCSAANSSHWYFDLWGRGTLVTVSS 575. PSMA-J
VH artificial nt
GAAGTGCAGCTGGTGCAGTCTGGCGCCGAAGTGAAGAAGCCCGGCGAGTCCCTGAAGATCTCCTG
CAAGGGCTCCGGGTACTCCTTCACCTCCTACTGGATCGGCTGGGCCAGGCAGATGCCAGGCAAGG
GCCTGGAATGGATGGGCATCATCTACCCTGGCGACTCCGACACCCGGTACTCTCCCAGCTTCCAG
GGCCAGGTGACCATCTCTGCCGACAAGTCCATCTCCACCGCCTACCTGCAGTGGTCCTCCCTGAA
GGCCTCCGACACCGCCATGTACTATTGCTCCGCCGCCAACTCCTCCCACTGGTACTTCGACCTGT
GGGGCAGAGGCACCCTGGTGACCGTGTCTTCC 576. PSMA-J HCDR1 artificial aa
SYWIG 577. PSMA-J HCDR2 artificial aa IIYPGDSDTRYSPSFQG 578. PSMA-J
HCDR3 artificial aa ANSSHWYFDL 579. PSMA-J LH artificial aa
EIVLTQSPATLSLSPGERATLSCRASQSVSSYLAWFQQKPGQAPRLLIYDASNRATGIPARFSGS
GSGTDFTLTISSLEPEDFAVYYCQQRSNWLMYTFGQGTKLEIKGGGGSGGGGSGGGGSEVQLVQS
GAEVKKPGESLKISCKGSGYSFTSYWIGWARQMPGKGLEWMGIIYPGDSDTRYSPSFQGQVTISA
DKSISTAYLQWSSLKASDTAMYYCSAANSSHWYFDLWGRGTLVTVSS 580. PSMA-J LH
artificial nt
GAGATCGTGCTGACCCAGTCCCCTGCCACCCTGTCTCTGTCTCCCGGCGAGAGAGCCACCCTGAG
CTGCCGGGCCTCCCAGTCCGTGTCCTCCTACCTGGCCTGGTTCCAGCAGAAGCCTGGACAGGCCC
CTAGGCTGCTGATCTACGACGCCTCCAACAGGGCTACCGGCATCCCTGCCCGGTTCTCCGGCTCT
GGCTCCGGCACCGACTTCACCCTGACCATCTCCAGCCTGGAACCTGAGGACTTCGCCGTGTACTA
CTGCCAGCAGCGGTCCAACTGGCTGATGTATACCTTCGGCCAGGGCACCAAGCTGGAAATCAAGG
GCGGAGGGGGATCTGGGGGCGGAGGAAGTGGAGGGGGCGGATCCGAAGTGCAGCTGGTGCAGTCT
GGCGCCGAAGTGAAGAAGCCCGGCGAGTCCCTGAAGATCTCCTGCAAGGGCTCCGGGTACTCCTT
CACCTCCTACTGGATCGGCTGGGCCAGGCAGATGCCAGGCAAGGGCCTGGAATGGATGGGCATCA
TCTACCCTGGCGACTCCGACACCCGGTACTCTCCCAGCTTCCAGGGCCAGGTGACCATCTCTGCC
GACAAGTCCATCTCCACCGCCTACCTGCAGTGGTCCTCCCTGAAGGCCTCCGACACCGCCATGTA
CTATTGCTCCGCCGCCAACTCCTCCCACTGGTACTTCGACCTGTGGGGCAGAGGCACCCTGGTGA
CCGTGTCTTCC 581. PSMA-J LH .times. I2C artificial aa
EIVLTQSPATLSLSPGERATLSCRASQSVSSYLAWFQQKPGQAPRLLIYDASNRATGIPARFSGS
HL
GSGTDFTLTISSLEPEDFAVYYCQQRSNWLMYTFGQGTKLEIKGGGGSGGGGSGGGGSEVQLVQS
GAEVKKPGESLKISCKGSGYSFTSYWIGWARQMPGKGLEWMGIIYPGDSDTRYSPSFQGQVTISA
DKSISTAYLQWSSLKASDTAMYYCSAANSSHWYFDLWGRGTLVTVSSGGGGSEVQLVESGGGLVQ
PGGSLKLSCAASGFTFNKYAMNWVRQAPGKGLEWVARIRSKYNNYATYYADSVKDRFTISRDDSK
NTAYLQMNNLKTEDTAVYYCVRHGNFGNSYISYWAYWGQGTLVTVSSGGGGSGGGGSGGGGSQTV
VTQEPSLTVSPGGTVTLTCGSSTGAVTSGNYPNWVQQKPGQAPRGLIGGTKFLAPGTPARFSGSL
LGGKAALTLSGVQPEDEAEYYCVLWYSNRWVFGGGTKLTVL 582. PSMA-J LH .times.
I2C artificial nt
GAGATCGTGCTGACCCAGTCCCCTGCCACCCTGTCTCTGTCTCCCGGCGAGAGAGCCACCCTGAG
HL
CTGCCGGGCCTCCCAGTCCGTGTCCTCCTACCTGGCCTGGTTCCAGCAGAAGCCTGGACAGGCCC
CTAGGCTGCTGATCTACGACGCCTCCAACAGGGCTACCGGCATCCCTGCCCGGTTCTCCGGCTCT
GGCTCCGGCACCGACTTCACCCTGACCATCTCCAGCCTGGAACCTGAGGACTTCGCCGTGTACTA
CTGCCAGCAGCGGTCCAACTGGCTGATGTATACCTTCGGCCAGGGCACCAAGCTGGAAATCAAGG
GCGGAGGGGGATCTGGGGGCGGAGGAAGTGGAGGGGGCGGATCCGAAGTGCAGCTGGTGCAGTCT
GGCGCCGAAGTGAAGAAGCCCGGCGAGTCCCTGAAGATCTCCTGCAAGGGCTCCGGGTACTCCTT
CACCTCCTACTGGATCGGCTGGGCCAGGCAGATGCCAGGCAAGGGCCTGGAATGGATGGGCATCA
TCTACCCTGGCGACTCCGACACCCGGTACTCTCCCAGCTTCCAGGGCCAGGTGACCATCTCTGCC
GACAAGTCCATCTCCACCGCCTACCTGCAGTGGTCCTCCCTGAAGGCCTCCGACACCGCCATGTA
CTATTGCTCCGCCGCCAACTCCTCCCACTGGTACTTCGACCTGTGGGGCAGAGGCACCCTGGTGA
CCGTGTCTTCCGGAGGTGGTGGATCCGAGGTGCAGCTGGTCGAGTCTGGAGGAGGATTGGTGCAG
CCTGGAGGGTCATTGAAACTCTCATGTGCAGCCTCTGGATTCACCTTCAATAAGTACGCCATGAA
CTGGGTCCGCCAGGCTCCAGGAAAGGGTTTGGAATGGGTTGCTCGCATAAGAAGTAAATATAATA
ATTATGCAACATATTATGCCGATTCAGTGAAAGACAGGTTCACCATCTCCAGAGATGATTCAAAA
AACACTGCCTATCTACAAATGAACAACTTGAAAACTGAGGACACTGCCGTGTACTACTGTGTGAG
ACATGGGAACTTCGGTAATAGCTACATATCCTACTGGGCTTACTGGGGCCAAGGGACTCTGGTCA
CCGTCTCCTCAGGTGGTGGTGGTTCTGGCGGCGGCGGCTCCGGTGGTGGTGGTTCTCAGACTGTT
GTGACTCAGGAACCTTCACTCACCGTATCACCTGGTGGAACAGTCACACTCACTTGTGGCTCCTC
GACTGGGGCTGTTACATCTGGCAACTACCCAAACTGGGTCCAACAAAAACCAGGTCAGGCACCCC
GTGGTCTAATAGGTGGGACTAAGTTCCTCGCCCCCGGTACTCCTGCCAGATTCTCAGGCTCCCTG
CTTGGAGGCAAGGCTGCCCTCACCCTCTCAGGGGTACAGCCAGAGGATGAGGCAGAATATTACTG
TGTTCTATGGTACAGCAACCGCTGGGTGTTCGGTGGAGGAACCAAACTGACTGTCCTA 583.
PSMA-L VL artificial aa
EIVLTQSPATLSLSPGERATLSCRASQSVSSYLAWYQQKPGQAPRLLIYDASNRATGIPARFSGS
GSGTDFTLTISSLEPEDFAVYYCQQRSDWLMYTFGQGTKLEIK 584. PSMA-L VL
artificial nt
GAGATCGTGCTGACCCAGTCCCCTGCCACCCTGTCTCTGTCTCCCGGCGAGAGAGCCACCCTGAG
CTGCCGGGCCTCCCAGTCCGTGTCCTCCTACCTGGCCTGGTATCAGCAGAAGCCTGGACAGGCCC
CTAGGCTGCTGATCTACGACGCCTCCAACAGGGCTACCGGCATCCCTGCCCGGTTCTCCGGCTCT
GGCTCCGGCACCGACTTCACCCTGACCATCTCCAGCCTGGAACCTGAGGACTTCGCCGTGTACTA
CTGCCAGCAGCGGTCCGACTGGCTGATGTATACCTTCGGCCAGGGCACCAAGCTGGAAATCAAG
585. PSMA-L LCDR1 artificial aa RASQSVSSYLA 586. PSMA-L LCDR2
artificial aa DASNRAT 587. PSMA-L LCDR3 artificial aa QQRSDWLMYT
588. PSMA-L VH artificial aa
EVQLVQSGAEVKTPGESLKISCKGSGYTFTSYWIGWVRQMPGKGPEWMGIIYPGDSDTRYSPSFQ
GQVTFSADKSISTAYLQWNSLKTSDTAMYYCATANPSYWYFDLWGRGTLVTVSS 589. PSMA-L
VH artificial nt
GAAGTGCAGCTGGTGCAGTCTGGCGCCGAAGTGAAAACCCCTGGCGAGTCCCTGAAGATCTCCTG
CAAGGGCTCCGGCTACACCTTCACCTCTTACTGGATCGGCTGGGTGCGCCAGATGCCTGGCAAGG
GCCCTGAGTGGATGGGCATCATCTACCCTGGCGACTCCGACACCCGGTACTCTCCCAGCTTCCAG
GGCCAGGTGACCTTCTCCGCCGACAAGTCCATCTCCACCGCCTACCTGCAGTGGAACTCCCTGAA
AACCTCCGACACCGCCATGTACTATTGCGCCACCGCCAACCCTAGCTACTGGTACTTCGACCTGT
GGGGCAGAGGCACCCTGGTGACCGTGTCTTCC 590. PSMA-L HCDR1 artificial aa
SYWIG 591. PSMA-L HCDR2 artificial aa IIYPGDSDTRYSPSFQG 592. PSMA-L
HCDR3 artificial aa ANPSYWYFDL 593. PSMA-L LH artificial aa
EIVLTQSPATLSLSPGERATLSCRASQSVSSYLAWYQQKPGQAPRLLIYDASNRATGIPARFSGS
GSGTDFTLTISSLEPEDFAVYYCQQRSDWLMYTFGQGTKLEIKGGGGSGGGGSGGGGSEVQLVQS
GAEVKTPGESLKISCKGSGYTFTSYWIGWVRQMPGKGPEWMGIIYPGDSDTRYSPSFQGQVTFSA
DKSISTAYLQWNSLKTSDTAMYYCATANPSYWYFDLWGRGTLVTVSS 594. PSMA-L LH
artificial nt
GAGATCGTGCTGACCCAGTCCCCTGCCACCCTGTCTCTGTCTCCCGGCGAGAGAGCCACCCTGAG
CTGCCGGGCCTCCCAGTCCGTGTCCTCCTACCTGGCCTGGTATCAGCAGAAGCCTGGACAGGCCC
CTAGGCTGCTGATCTACGACGCCTCCAACAGGGCTACCGGCATCCCTGCCCGGTTCTCCGGCTCT
GGCTCCGGCACCGACTTCACCCTGACCATCTCCAGCCTGGAACCTGAGGACTTCGCCGTGTACTA
CTGCCAGCAGCGGTCCGACTGGCTGATGTATACCTTCGGCCAGGGCACCAAGCTGGAAATCAAGG
GCGGAGGGGGATCTGGCGGCGGAGGAAGTGGAGGGGGCGGATCCGAAGTGCAGCTGGTGCAGTCT
GGCGCCGAAGTGAAAACCCCTGGCGAGTCCCTGAAGATCTCCTGCAAGGGCTCCGGCTACACCTT
CACCTCTTACTGGATCGGCTGGGTGCGCCAGATGCCTGGCAAGGGCCCTGAGTGGATGGGCATCA
TCTACCCTGGCGACTCCGACACCCGGTACTCTCCCAGCTTCCAGGGCCAGGTGACCTTCTCCGCC
GACAAGTCCATCTCCACCGCCTACCTGCAGTGGAACTCCCTGAAAACCTCCGACACCGCCATGTA
CTATTGCGCCACCGCCAACCCTAGCTACTGGTACTTCGACCTGTGGGGCAGAGGCACCCTGGTGA
CCGTGTCTTCC 595. PSMA-L LH .times. I2C artificial aa
EIVLTQSPATLSLSPGERATLSCRASQSVSSYLAWYQQKPGQAPRLLIYDASNRATGIPARFSGS
HL
GSGTDFTLTISSLEPEDFAVYYCQQRSDWLMYTFGQGTKLEIKGGGGSGGGGSGGGGSEVQLVQS
GAEVKTPGESLKISCKGSGYTFTSYWIGWVRQMPGKGPEWMGIIYPGDSDTRYSPSFQGQVTFSA
DKSISTAYLQWNSLKTSDTAMYYCATANPSYWYFDLWGRGTLVTVSSGGGGSEVQLVESGGGLVQ
PGGSLKLSCAASGFTFNKYAMNWVRQAPGKGLEWVARIRSKYNNYATYYADSVKDRFTISRDDSK
NTAYLQMNNLKTEDTAVYYCVRHGNFGNSYISYWAYWGQGTLVTVSSGGGGSGGGGSGGGGSQTV
VTQEPSLTVSPGGTVTLTCGSSTGAVTSGNYPNWVQQKPGQAPRGLIGGTKFLAPGTPARFSGSL
LGGKAALTLSGVQPEDEAEYYCVLWYSNRWVFGGGTKLTVL 596. PSMA-L LH .times.
I2C artificial nt
GAGATCGTGCTGACCCAGTCCCCTGCCACCCTGTCTCTGTCTCCCGGCGAGAGAGCCACCCTGAG
HL
CTGCCGGGCCTCCCAGTCCGTGTCCTCCTACCTGGCCTGGTATCAGCAGAAGCCTGGACAGGCCC
CTAGGCTGCTGATCTACGACGCCTCCAACAGGGCTACCGGCATCCCTGCCCGGTTCTCCGGCTCT
GGCTCCGGCACCGACTTCACCCTGACCATCTCCAGCCTGGAACCTGAGGACTTCGCCGTGTACTA
CTGCCAGCAGCGGTCCGACTGGCTGATGTATACCTTCGGCCAGGGCACCAAGCTGGAAATCAAGG
GCGGAGGGGGATCTGGCGGCGGAGGAAGTGGAGGGGGCGGATCCGAAGTGCAGCTGGTGCAGTCT
GGCGCCGAAGTGAAAACCCCTGGCGAGTCCCTGAAGATCTCCTGCAAGGGCTCCGGCTACACCTT
CACCTCTTACTGGATCGGCTGGGTGCGCCAGATGCCTGGCAAGGGCCCTGAGTGGATGGGCATCA
TCTACCCTGGCGACTCCGACACCCGGTACTCTCCCAGCTTCCAGGGCCAGGTGACCTTCTCCGCC
GACAAGTCCATCTCCACCGCCTACCTGCAGTGGAACTCCCTGAAAACCTCCGACACCGCCATGTA
CTATTGCGCCACCGCCAACCCTAGCTACTGGTACTTCGACCTGTGGGGCAGAGGCACCCTGGTGA
CCGTGTCTTCCGGAGGTGGTGGATCCGAGGTGCAGCTGGTCGAGTCTGGAGGAGGATTGGTGCAG
CCTGGAGGGTCATTGAAACTCTCATGTGCAGCCTCTGGATTCACCTTCAATAAGTACGCCATGAA
CTGGGTCCGCCAGGCTCCAGGAAAGGGTTTGGAATGGGTTGCTCGCATAAGAAGTAAATATAATA
ATTATGCAACATATTATGCCGATTCAGTGAAAGACAGGTTCACCATCTCCAGAGATGATTCAAAA
AACACTGCCTATCTACAAATGAACAACTTGAAAACTGAGGACACTGCCGTGTACTACTGTGTGAG
ACATGGGAACTTCGGTAATAGCTACATATCCTACTGGGCTTACTGGGGCCAAGGGACTCTGGTCA
CCGTCTCCTCAGGTGGTGGTGGTTCTGGCGGCGGCGGCTCCGGTGGTGGTGGTTCTCAGACTGTT
GTGACTCAGGAACCTTCACTCACCGTATCACCTGGTGGAACAGTCACACTCACTTGTGGCTCCTC
GACTGGGGCTGTTACATCTGGCAACTACCCAAACTGGGTCCAACAAAAACCAGGTCAGGCACCCC
GTGGTCTAATAGGTGGGACTAAGTTCCTCGCCCCCGGTACTCCTGCCAGATTCTCAGGCTCCCTG
CTTGGAGGCAAGGCTGCCCTCACCCTCTCAGGGGTACAGCCAGAGGATGAGGCAGAATATTACTG
TGTTCTATGGTACAGCAACCGCTGGGTGTTCGGTGGAGGAACCAAACTGACTGTCCTA 597. PM
99-A8 VL artificial aa
DIQMTQSPSTLAASAGEKVTITCRASSSVTYMHWYQQKPGKAPKLLIYDTSKVASGVPSRFSGSG
SGTAFTLTISSVQTDDFATYYCQQWSRNSPYTFGQGTKLEIK 598. PM 99-A8 VL
artificial nt
GACATCCAGATGACTCAGTCTCCATCAACCCTGGCTGCATCTGCGGGGGAGAAAGTCACCATCAC
CTGCCGTGCCAGCTCCAGTGTGACTTACATGCACTGGTACCAGCAGAAGCCAGGCAAAGCCCCTA
AATTATTGATTTATGACACATCCAAAGTGGCTTCTGGAGTCCCAAGTCGCTTCAGTGGCAGTGGG
TCTGGGACCGCATTTACTCTCACAATCAGCTCCGTGCAGACTGATGACTTTGCCACTTATTACTG
TCAGCAGTGGAGTAGGAACTCACCCTACACGTTTGGACAGGGGACCAAGCTGGAAATCAAA 599.
PM 99-A8 LCDR1 artificial aa RASSSVTYMH 600. PM 99-A8 LCDR2
artificial aa DTSKVAS 601. PM 99-A8 LCDR3 artificial aa QQWSRNSPYT
602. PM 99-A8 VH artificial aa
EVQLLESGGGLVQPGGSLRLSCAASGFTFSDFFMAWVRQAPGKGLEWVATIVSDGGSTYYRDSVK
GRFTISRDNAKNTLYLQMDSLRAEDTAVYYCAKRGNSGYYVMDAWGQGTTVTVSS 603. PM
99-A8 VH artificial nt
GAGGTGCAGCTGCTCGAGTCTGGTGGAGGCTTAGTGCAGCCTGGAGGGTCCCTAAGACTCTCCTG
TGCAGCCTCAGGATTCACTTTCAGTGACTTTTTCATGGCCTGGGTCCGCCAGGCTCCAGGGAAGG
GGCTGGAGTGGGTCGCAACCATTGTTTCTGATGGTGGTAGCACTTACTATCGCGACTCCGTGAAG
GGCCGTTTCACTATCTCCAGAGATAATGCAAAAAACACCCTGTACCTGCAAATGGACAGTCTGAG
GGCTGAGGACACGGCCGTTTATTACTGTGCAAAACGCGGCAATTCGGGGTACTATGTTATGGATG
CCTGGGGTCAAGGAACTACGGTCACCGTCTCCTCA 604. PM 99-A8 HCDR1 artificial
aa DFFMA 605. PM 99-A8 HCDR2 artificial aa TIVSDGGSTYYRDSVKG 606.
PM 99-A8 HCDR3 artificial aa RGNSGYYVMDA 607. PM 99-A8 HL
artificial aa
EVQLLESGGGLVQPGGSLRLSCAASGFTFSDFFMAWVRQAPGKGLEWVATIVSDGGSTYYRDSVK
GRFTISRDNAKNTLYLQMDSLRAEDTAVYYCAKRGNSGYYVMDAWGQGTTVTVSSGGGGSGGGGS
GGGGSDIQMTQSPSTLAASAGEKVTITCRASSSVTYMHWYQQKPGKAPKLLIYDTSKVASGVPSR
FSGSGSGTAFTLTISSVQTDDFATYYCQQWSRNSPYTFGQGTKLEIK 608. PM 99-A8 HL
artificial nt
GAGGTGCAGCTGCTCGAGTCTGGTGGAGGCTTAGTGCAGCCTGGAGGGTCCCTAAGACTCTCCTG
TGCAGCCTCAGGATTCACTTTCAGTGACTTTTTCATGGCCTGGGTCCGCCAGGCTCCAGGGAAGG
GGCTGGAGTGGGTCGCAACCATTGTTTCTGATGGTGGTAGCACTTACTATCGCGACTCCGTGAAG
GGCCGTTTCACTATCTCCAGAGATAATGCAAAAAACACCCTGTACCTGCAAATGGACAGTCTGAG
GGCTGAGGACACGGCCGTTTATTACTGTGCAAAACGCGGCAATTCGGGGTACTATGTTATGGATG
CCTGGGGTCAAGGAACTACGGTCACCGTCTCCTCAGGTGGTGGTGGTTCTGGCGGCGGCGGCTCC
GGTGGTGGTGGTTCTGACATCCAGATGACTCAGTCTCCATCAACCCTGGCTGCATCTGCGGGGGA
GAAAGTCACCATCACCTGCCGTGCCAGCTCCAGTGTGACTTACATGCACTGGTACCAGCAGAAGC
CAGGCAAAGCCCCTAAATTATTGATTTATGACACATCCAAAGTGGCTTCTGGAGTCCCAAGTCGC
TTCAGTGGCAGTGGGTCTGGGACCGCATTTACTCTCACAATCAGCTCCGTGCAGACTGATGACTT
TGCCACTTATTACTGTCAGCAGTGGAGTAGGAACTCACCCTACACGTTTGGACAGGGGACCAAGC
TGGAAATCAAA 609. PM 99-A8 HL .times. I2C artificial aa
EVQLLESGGGLVQPGGSLRLSCAASGFTFSDFFMAWVRQAPGKGLEWVATIVSDGGSTYYRDSVK
HL
GRFTISRDNAKNTLYLQMDSLRAEDTAVYYCAKRGNSGYYVMDAWGQGTTVTVSSGGGGSGGGGS
GGGGSDIQMTQSPSTLAASAGEKVTITCRASSSVTYMHWYQQKPGKAPKLLIYDTSKVASGVPSR
FSGSGSGTAFTLTISSVQTDDFATYYCQQWSRNSPYTFGQGTKLEIKSGGGGSEVQLVESGGGLV
QPGGSLKLSCAASGFTFNKYAMNWVRQAPGKGLEWVARIRSKYNNYATYYADSVKDRFTISRDDS
KNTAYLQMNNLKTEDTAVYYCVRHGNFGNSYISYWAYWGQGTLVTVSSGGGGSGGGGSGGGGSQT
VVTQEPSLTVSPGGTVTLTCGSSTGAVTSGNYPNWVQQKPGQAPRGLIGGTKFLAPGTPARFSGS
LLGGKAALTLSGVQPEDEAEYYCVLWYSNRWVFGGGTKLTVL 610. PM 99-A8 HL .times.
I2C artificial nt
GAGGTGCAGCTGCTCGAGTCTGGTGGAGGCTTAGTGCAGCCTGGAGGGTCCCTAAGACTCTCCTG
HL
TGCAGCCTCAGGATTCACTTTCAGTGACTTTTTCATGGCCTGGGTCCGCCAGGCTCCAGGGAAGG
GGCTGGAGTGGGTCGCAACCATTGTTTCTGATGGTGGTAGCACTTACTATCGCGACTCCGTGAAG
GGCCGTTTCACTATCTCCAGAGATAATGCAAAAAACACCCTGTACCTGCAAATGGACAGTCTGAG
GGCTGAGGACACGGCCGTTTATTACTGTGCAAAACGCGGCAATTCGGGGTACTATGTTATGGATG
CCTGGGGTCAAGGAACTACGGTCACCGTCTCCTCAGGTGGTGGTGGTTCTGGCGGCGGCGGCTCC
GGTGGTGGTGGTTCTGACATCCAGATGACTCAGTCTCCATCAACCCTGGCTGCATCTGCGGGGGA
GAAAGTCACCATCACCTGCCGTGCCAGCTCCAGTGTGACTTACATGCACTGGTACCAGCAGAAGC
CAGGCAAAGCCCCTAAATTATTGATTTATGACACATCCAAAGTGGCTTCTGGAGTCCCAAGTCGC
TTCAGTGGCAGTGGGTCTGGGACCGCATTTACTCTCACAATCAGCTCCGTGCAGACTGATGACTT
TGCCACTTATTACTGTCAGCAGTGGAGTAGGAACTCACCCTACACGTTTGGACAGGGGACCAAGC
TGGAAATCAAATCCGGAGGTGGTGGATCCGAGGTGCAGCTGGTCGAGTCTGGAGGAGGATTGGTG
CAGCCTGGAGGGTCATTGAAACTCTCATGTGCAGCCTCTGGATTCACCTTCAATAAGTACGCCAT
GAACTGGGTCCGCCAGGCTCCAGGAAAGGGTTTGGAATGGGTTGCTCGCATAAGAAGTAAATATA
ATAATTATGCAACATATTATGCCGATTCAGTGAAAGACAGGTTCACCATCTCCAGAGATGATTCA
AAAAACACTGCCTATCTACAAATGAACAACTTGAAAACTGAGGACACTGCCGTGTACTACTGTGT
GAGACATGGGAACTTCGGTAATAGCTACATATCCTACTGGGCTTACTGGGGCCAAGGGACTCTGG
TCACCGTCTCCTCAGGTGGTGGTGGTTCTGGCGGCGGCGGCTCCGGTGGTGGTGGTTCTCAGACT
GTTGTGACTCAGGAACCTTCACTCACCGTATCACCTGGTGGAACAGTCACACTCACTTGTGGCTC
CTCGACTGGGGCTGTTACATCTGGCAACTACCCAAACTGGGTCCAACAAAAACCAGGTCAGGCAC
CCCGTGGTCTAATAGGTGGGACTAAGTTCCTCGCCCCCGGTACTCCTGCCAGATTCTCAGGCTCC
CTGCTTGGAGGCAAGGCTGCCCTCACCCTCTCAGGGGTACAGCCAGAGGATGAGGCAGAATATTA
CTGTGTTCTATGGTACAGCAACCGCTGGGTGTTCGGTGGAGGAACCAAACTGACTGTCCTA 611.
PM 86-A10 VL artificial aa
DIQMTQSPSTLSASVGDRVTITCRASSSVTYMHWYQQKPGKAPKLLIYDTSKVASGVPSRFSGSG
SGTEFTLTISSPQPDDFATYYCQQWSRNSPYTFGQGTKLEIK 612. PM 86-A10 VL
artificial nt
GACATCCAGATGACTCAGTCTCCATCAACCCTGTCTGCATCTGTGGGGGACAGAGTCACCATCAC
CTGCCGGGCCAGCTCCAGTGTGACTTACATGCACTGGTACCAGCAGAAGCCAGGCAAAGCCCCTA
AATTATTGATTTATGACACATCCAAAGTGGCTTCTGGAGTCCCAAGTCGCTTCAGTGGCAGTGGG
TCTGGGACCGAATTTACTCTCACAATCAGCTCCCCGCAGCCTGATGACTTTGCCACTTATTACTG
TCAGCAGTGGAGTAGGAACTCACCCTACACGTTTGGGCAGGGGACCAAGCTGGAAATCAAA 613.
PM 86-A10 LCDR1 artificial aa RASSSVTYMH 614. PM 86-A10 LCDR2
artificial aa DTSKVAS 615. PM 86-A10 LCDR3 artificial aa QQWSRNSPYT
616. PM 86-A10 VH artificial aa
EVQLLESDGGLVQPGGSLKLSCAASGFTFSDFFMAWVRQAPGKGLEWVATIVSDGGSTYYRDSVK
GRFTISRDNSKNTLYLQMNNLRSEDTAVYYCAKRGNSGYYVMDAWGQGTTVTVSS 617. PM
86-A10 VH artificial nt
GAGGTGCAGCTGCTCGAGTCTGATGGAGGCTTAGTGCAGCCTGGAGGGTCCCTAAAACTCTCCTG
TGCAGCCTCAGGATTCACTTTCAGTGACTTTTTCATGGCCTGGGTCCGCCAGGCTCCAGGGAAGG
GGCTGGAGTGGGTCGCAACCATTGTTTCTGATGGTGGTAGCACTTACTATCGCGACTCCGTGAAG
GGCCGTTTCACTATCTCCAGAGATAATTCAAAAAACACCCTGTACCTGCAAATGAACAATCTGAG
GTCTGAGGACACGGCCGTTTATTACTGTGCAAAACGCGGCAATTCGGGGTACTATGTTATGGATG
CCTGGGGTCAAGGAACTACGGTCACCGTCTCCTCA 618. PM 86-A10 HCDR1 artificial
aa DFFMA 619. PM 86-A10 HCDR2 artificial aa TIVSDGGSTYYRDSVKG 620.
PM 86-A10 HCDR3 artificial aa RGNSGYYVMDA 621. PM 86-A10 HL
artificial aa
EVQLLESDGGLVQPGGSLKLSCAASGFTFSDFFMAWVRQAPGKGLEWVATIVSDGGSTYYRDSVK
GRFTISRDNSKNTLYLQMNNLRSEDTAVYYCAKRGNSGYYVMDAWGQGTTVTVSSGGGGSGGGGS
GGGGSDIQMTQSPSTLSASVGDRVTITCRASSSVTYMHWYQQKPGKAPKLLIYDTSKVASGVPSR
FSGSGSGTEFTLTISSPQPDDFATYYCQQWSRNSPYTFGQGTKLEIK 622. PM 86-A10 HL
artificial nt
GAGGTGCAGCTGCTCGAGTCTGATGGAGGCTTAGTGCAGCCTGGAGGGTCCCTAAAACTCTCCTG
TGCAGCCTCAGGATTCACTTTCAGTGACTTTTTCATGGCCTGGGTCCGCCAGGCTCCAGGGAAGG
GGCTGGAGTGGGTCGCAACCATTGTTTCTGATGGTGGTAGCACTTACTATCGCGACTCCGTGAAG
GGCCGTTTCACTATCTCCAGAGATAATTCAAAAAACACCCTGTACCTGCAAATGAACAATCTGAG
GTCTGAGGACACGGCCGTTTATTACTGTGCAAAACGCGGCAATTCGGGGTACTATGTTATGGATG
CCTGGGGTCAAGGAACTACGGTCACCGTCTCCTCAGGTGGTGGTGGTTCTGGCGGCGGCGGCTCC
GGTGGTGGTGGTTCTGACATCCAGATGACTCAGTCTCCATCAACCCTGTCTGCATCTGTGGGGGA
CAGAGTCACCATCACCTGCCGGGCCAGCTCCAGTGTGACTTACATGCACTGGTACCAGCAGAAGC
CAGGCAAAGCCCCTAAATTATTGATTTATGACACATCCAAAGTGGCTTCTGGAGTCCCAAGTCGC
TTCAGTGGCAGTGGGTCTGGGACCGAATTTACTCTCACAATCAGCTCCCCGCAGCCTGATGACTT
TGCCACTTATTACTGTCAGCAGTGGAGTAGGAACTCACCCTACACGTTTGGGCAGGGGACCAAGC
TGGAAATCAAA 623. PM 86-A10 HL .times. artificial aa
EVQLLESDGGLVQPGGSLKLSCAASGFTFSDFFMAWVRQAPGKGLEWVATIVSDGGSTYYRDSVK
I2C HL
GRFTISRDNSKNTLYLQMNNLRSEDTAVYYCAKRGNSGYYVMDAWGQGTTVTVSSGGGGSGGGG- S
GGGGSDIQMTQSPSTLSASVGDRVTITCRASSSVTYMHWYQQKPGKAPKLLIYDTSKVASGVPSR
FSGSGSGTEFTLTISSPQPDDFATYYCQQWSRNSPYTFGQGTKLEIKSGGGGSEVQLVESGGGLV
QPGGSLKLSCAASGFTFNKYAMNWVRQAPGKGLEWVARIRSKYNNYATYYADSVKDRFTISRDDS
KNTAYLQMNNLKTEDTAVYYCVRHGNFGNSYISYWAYWGQGTLVTVSSGGGGSGGGGSGGGGSQT
VVTQEPSLTVSPGGTVTLTCGSSTGAVTSGNYPNWVQQKPGQAPRGLIGGTKFLAPGTPARFSGS
LLGGKAALTLSGVQPEDEAEYYCVLWYSNRWVFGGGTKLTVL 624. PM 86-A10 HL
.times. artificial nt
GAGGTGCAGCTGCTCGAGTCTGATGGAGGCTTAGTGCAGCCTGGAGGGTCCCTAAAACTCTCCTG
I2C HL
TGCAGCCTCAGGATTCACTTTCAGTGACTTTTTCATGGCCTGGGTCCGCCAGGCTCCAGGGAAG- G
GGCTGGAGTGGGTCGCAACCATTGTTTCTGATGGTGGTAGCACTTACTATCGCGACTCCGTGAAG
GGCCGTTTCACTATCTCCAGAGATAATTCAAAAAACACCCTGTACCTGCAAATGAACAATCTGAG
GTCTGAGGACACGGCCGTTTATTACTGTGCAAAACGCGGCAATTCGGGGTACTATGTTATGGATG
CCTGGGGTCAAGGAACTACGGTCACCGTCTCCTCAGGTGGTGGTGGTTCTGGCGGCGGCGGCTCC
GGTGGTGGTGGTTCTGACATCCAGATGACTCAGTCTCCATCAACCCTGTCTGCATCTGTGGGGGA
CAGAGTCACCATCACCTGCCGGGCCAGCTCCAGTGTGACTTACATGCACTGGTACCAGCAGAAGC
CAGGCAAAGCCCCTAAATTATTGATTTATGACACATCCAAAGTGGCTTCTGGAGTCCCAAGTCGC
TTCAGTGGCAGTGGGTCTGGGACCGAATTTACTCTCACAATCAGCTCCCCGCAGCCTGATGACTT
TGCCACTTATTACTGTCAGCAGTGGAGTAGGAACTCACCCTACACGTTTGGGCAGGGGACCAAGC
TGGAAATCAAATCCGGAGGTGGTGGATCCGAGGTGCAGCTGGTCGAGTCTGGAGGAGGATTGGTG
CAGCCTGGAGGGTCATTGAAACTCTCATGTGCAGCCTCTGGATTCACCTTCAATAAGTACGCCAT
GAACTGGGTCCGCCAGGCTCCAGGAAAGGGTTTGGAATGGGTTGCTCGCATAAGAAGTAAATATA
ATAATTATGCAACATATTATGCCGATTCAGTGAAAGACAGGTTCACCATCTCCAGAGATGATTCA
AAAAACACTGCCTATCTACAAATGAACAACTTGAAAACTGAGGACACTGCCGTGTACTACTGTGT
GAGACATGGGAACTTCGGTAATAGCTACATATCCTACTGGGCTTACTGGGGCCAAGGGACTCTGG
TCACCGTCTCCTCAGGTGGTGGTGGTTCTGGCGGCGGCGGCTCCGGTGGTGGTGGTTCTCAGACT
GTTGTGACTCAGGAACCTTCACTCACCGTATCACCTGGTGGAACAGTCACACTCACTTGTGGCTC
CTCGACTGGGGCTGTTACATCTGGCAACTACCCAAACTGGGTCCAACAAAAACCAGGTCAGGCAC
CCCGTGGTCTAATAGGTGGGACTAAGTTCCTCGCCCCCGGTACTCCTGCCAGATTCTCAGGCTCC
CTGCTTGGAGGCAAGGCTGCCCTCACCCTCTCAGGGGTACAGCCAGAGGATGAGGCAGAATATTA
CTGTGTTCTATGGTACAGCAACCGCTGGGTGTTCGGTGGAGGAACCAAACTGACTGTCCTA 625.
PM 86-B4-2 VL artificial aa
DIQMTQSPSTLAASPGDRVTITCRASSSVTYMHWYQQKPGKSPKLLIYDTSKVASGVPNRFSGSG
SGTEFTLTISSLQPDDIATYYCQQWSRNSPYTFGQGTKLEIK 626. PM 86-B4-2 VL
artificial nt
GACATCCAGATGACTCAGTCTCCATCAACCCTGGCTGCATCTCCGGGGGACAGAGTCACCATCAC
CTGCCGTGCCAGCTCCAGTGTGACTTACATGCACTGGTACCAGCAGAAGCCAGGCAAATCCCCTA
AATTATTGATTTATGACACATCCAAAGTGGCTTCTGGAGTCCCAAATCGCTTCAGTGGCAGTGGG
TCTGGGACCGAATTTACTCTCACAATCAGCTCCCTGCAGCCTGATGACATTGCCACTTATTACTG
TCAGCAGTGGAGTAGGAACTCACCCTACACGTTTGGACAGGGGACCAAGCTGGAAATCAAA 627.
PM 86-B4-2 LCDR1 artificial aa RASSSVTYMH 628. PM 86-B4-2 LCDR2
artificial aa DTSKVAS 629. PM 86-B4-2 LCDR3 artificial aa
QQWSRNSPYT 630. PM 86-B4-2 VH artificial aa
EVQLLESDGGLVQPGGSLRLSCAASGFTFSDFFMAWVRQAPGKGLEWVATIVSDGGSTYYRDSVK
GRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAKRGNSGYYVMDAWGQGTTVTVSS 631. PM
86-B4-2 VH artificial nt
GAGGTGCAGCTGCTCGAGTCTGATGGAGGCTTAGTGCAGCCTGGAGGGTCCCTAAGACTCTCCTG
TGCAGCCTCAGGATTCACTTTCAGTGACTTTTTCATGGCCTGGGTCCGCCAGGCTCCAGGGAAGG
GGCTGGAGTGGGTCGCAACCATTGTTTCTGATGGTGGTAGCACTTACTATCGCGACTCCGTGAAG
GGCCGTTTCACTATCTCCAGAGATAATTCAAAAAACACCCTGTACCTGCAAATGAACAGTCTGAG
GGCTGAGGACACGGCCGTTTATTACTGTGCAAAACGCGGCAATTCGGGGTACTATGTTATGGATG
CCTGGGGTCAAGGAACTACGGTCACCGTCTCCTCA 632. PM 86-B4-2 HCDR1
artificial aa DFFMA 633. PM 86-B4-2 HCDR2 artificial aa
TIVSDGGSTYYRDSVKG 634. PM 86-B4-2 HCDR3 artificial aa RGNSGYYVMDA
635. PM 86-B4-2 HL artificial aa
EVQLLESDGGLVQPGGSLRLSCAASGFTFSDFFMAWVRQAPGKGLEWVATIVSDGGSTYYRDSVK
GRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAKRGNSGYYVMDAWGQGTTVTVSSGGGGSGGGGS
GGGGSDIQMTQSPSTLAASPGDRVTITCRASSSVTYMHWYQQKPGKSPKLLIYDTSKVASGVPNR
FSGSGSGTEFTLTISSLQPDDIATYYCQQWSRNSPYTFGQGTKLEIK 636. PM 86-B4-2 HL
artificial nt
GAGGTGCAGCTGCTCGAGTCTGATGGAGGCTTAGTGCAGCCTGGAGGGTCCCTAAGACTCTCCTG
TGCAGCCTCAGGATTCACTTTCAGTGACTTTTTCATGGCCTGGGTCCGCCAGGCTCCAGGGAAGG
GGCTGGAGTGGGTCGCAACCATTGTTTCTGATGGTGGTAGCACTTACTATCGCGACTCCGTGAAG
GGCCGTTTCACTATCTCCAGAGATAATTCAAAAAACACCCTGTACCTGCAAATGAACAGTCTGAG
GGCTGAGGACACGGCCGTTTATTACTGTGCAAAACGCGGCAATTCGGGGTACTATGTTATGGATG
CCTGGGGTCAAGGAACTACGGTCACCGTCTCCTCAGGTGGTGGTGGTTCTGGCGGCGGCGGCTCC
GGTGGTGGTGGTTCTGACATCCAGATGACTCAGTCTCCATCAACCCTGGCTGCATCTCCGGGGGA
CAGAGTCACCATCACCTGCCGTGCCAGCTCCAGTGTGACTTACATGCACTGGTACCAGCAGAAGC
CAGGCAAATCCCCTAAATTATTGATTTATGACACATCCAAAGTGGCTTCTGGAGTCCCAAATCGC
TTCAGTGGCAGTGGGTCTGGGACCGAATTTACTCTCACAATCAGCTCCCTGCAGCCTGATGACAT
TGCCACTTATTACTGTCAGCAGTGGAGTAGGAACTCACCCTACACGTTTGGACAGGGGACCAAGC
TGGAAATCAAA 637. PM 86-B4-2 HL .times. artificial aa
EVQLLESDGGLVQPGGSLRLSCAASGFTFSDFFMAWVRQAPGKGLEWVATIVSDGGSTYYRDSVK
I2C HL
GRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAKRGNSGYYVMDAWGQGTTVTVSSGGGGSGGGG- S
GGGGSDIQMTQSPSTLAASPGDRVTITCRASSSVTYMHWYQQKPGKSPKLLIYDTSKVASGVPNR
FSGSGSGTEFTLTISSLQPDDIATYYCQQWSRNSPYTFGQGTKLEIKSGGGGSEVQLVESGGGLV
QPGGSLKLSCAASGFTFNKYAMNWVRQAPGKGLEWVARIRSKYNNYATYYADSVKDRFTISRDDS
KNTAYLQMNNLKTEDTAVYYCVRHGNFGNSYISYWAYWGQGTLVTVSSGGGGSGGGGSGGGGSQT
VVTQEPSLTVSPGGTVTLTCGSSTGAVTSGNYPNWVQQKPGQAPRGLIGGTKFLAPGTPARFSGS
LLGGKAALTLSGVQPEDEAEYYCVLWYSNRWVFGGGTKLTVL 638. PM 86-B4-2 HL
.times. artificial nt
GAGGTGCAGCTGCTCGAGTCTGATGGAGGCTTAGTGCAGCCTGGAGGGTCCCTAAGACTCTCCTG
I2C HL
TGCAGCCTCAGGATTCACTTTCAGTGACTTTTTCATGGCCTGGGTCCGCCAGGCTCCAGGGAAG- G
GGCTGGAGTGGGTCGCAACCATTGTTTCTGATGGTGGTAGCACTTACTATCGCGACTCCGTGAAG
GGCCGTTTCACTATCTCCAGAGATAATTCAAAAAACACCCTGTACCTGCAAATGAACAGTCTGAG
GGCTGAGGACACGGCCGTTTATTACTGTGCAAAACGCGGCAATTCGGGGTACTATGTTATGGATG
CCTGGGGTCAAGGAACTACGGTCACCGTCTCCTCAGGTGGTGGTGGTTCTGGCGGCGGCGGCTCC
GGTGGTGGTGGTTCTGACATCCAGATGACTCAGTCTCCATCAACCCTGGCTGCATCTCCGGGGGA
CAGAGTCACCATCACCTGCCGTGCCAGCTCCAGTGTGACTTACATGCACTGGTACCAGCAGAAGC
CAGGCAAATCCCCTAAATTATTGATTTATGACACATCCAAAGTGGCTTCTGGAGTCCCAAATCGC
TTCAGTGGCAGTGGGTCTGGGACCGAATTTACTCTCACAATCAGCTCCCTGCAGCCTGATGACAT
TGCCACTTATTACTGTCAGCAGTGGAGTAGGAACTCACCCTACACGTTTGGACAGGGGACCAAGC
TGGAAATCAAATCCGGAGGTGGTGGATCCGAGGTGCAGCTGGTCGAGTCTGGAGGAGGATTGGTG
CAGCCTGGAGGGTCATTGAAACTCTCATGTGCAGCCTCTGGATTCACCTTCAATAAGTACGCCAT
GAACTGGGTCCGCCAGGCTCCAGGAAAGGGTTTGGAATGGGTTGCTCGCATAAGAAGTAAATATA
ATAATTATGCAACATATTATGCCGATTCAGTGAAAGACAGGTTCACCATCTCCAGAGATGATTCA
AAAAACACTGCCTATCTACAAATGAACAACTTGAAAACTGAGGACACTGCCGTGTACTACTGTGT
GAGACATGGGAACTTCGGTAATAGCTACATATCCTACTGGGCTTACTGGGGCCAAGGGACTCTGG
TCACCGTCTCCTCAGGTGGTGGTGGTTCTGGCGGCGGCGGCTCCGGTGGTGGTGGTTCTCAGACT
GTTGTGACTCAGGAACCTTCACTCACCGTATCACCTGGTGGAACAGTCACACTCACTTGTGGCTC
CTCGACTGGGGCTGTTACATCTGGCAACTACCCAAACTGGGTCCAACAAAAACCAGGTCAGGCAC
CCCGTGGTCTAATAGGTGGGACTAAGTTCCTCGCCCCCGGTACTCCTGCCAGATTCTCAGGCTCC
CTGCTTGGAGGCAAGGCTGCCCTCACCCTCTCAGGGGTACAGCCAGAGGATGAGGCAGAATATTA
CTGTGTTCTATGGTACAGCAACCGCTGGGTGTTCGGTGGAGGAACCAAACTGACTGTCCTA 639.
PM 98-B4 VL artificial aa
DIQMTQSPSTLSASVGEKVTITCRASSSVTYMHWYQQKPGKSPKLLIYDTSKVASGVPSRFSGSG
SGTAYTLTISSLQTDDFATYYCQQWSRNSPYTFGQGTKLEIK 640. PM 98-B4 VL
artificial nt
GACATCCAGATGACTCAGTCTCCATCAACCCTGTCTGCATCTGTGGGGGAGAAAGTCACCATCAC
CTGCCGTGCCAGCTCCAGTGTGACTTACATGCACTGGTACCAGCAGAAGCCAGGCAAATCCCCTA
AATTATTGATTTATGACACATCCAAAGTGGCTTCTGGAGTCCCAAGTCGCTTCAGTGGCAGTGGG
TCTGGGACCGCATATACTCTCACAATCAGCTCCCTGCAGACTGATGACTTTGCCACTTATTACTG
TCAGCAGTGGAGTAGGAACTCACCCTACACGTTTGGACAGGGGACCAAGCTGGAAATCAAA 641.
PM 98-B4 LCDR1 artificial aa RASSSVTYMH 642. PM 98-B4 LCDR2
artificial aa DTSKVAS 643. PM 98-B4 LCDR3 artificial aa QQWSRNSPYT
644. PM 98-B4 VH artificial aa
EVQLLESGGGLVQPGGSLRLSCAASGFTFSDFFMAWIRQAPGKGLEWVSTIVSDGGSTYYRDSVK
GRFTISRDNSQSTLYLQMDSLTAEDTAVYYCAKRGNSGYYVMDAWGQGTTVTVSS 645. PM
98-B4 VH artificial nt
GAGGTGCAGCTGCTCGAGTCTGGTGGAGGCTTAGTGCAGCCTGGAGGGTCCCTAAGACTCTCCTG
TGCAGCCTCAGGATTCACTTTCAGTGACTTTTTCATGGCCTGGATCCGCCAGGCTCCAGGGAAGG
GGCTGGAGTGGGTCTCAACCATTGTTTCTGATGGTGGTAGCACTTACTATCGCGACTCCGTGAAG
GGCCGTTTCACTATCTCCAGAGATAATTCACAAAGCACCCTGTACCTGCAAATGGACAGTCTGAC
GGCTGAGGACACGGCCGTTTATTACTGTGCAAAACGCGGCAATTCGGGGTACTATGTTATGGATG
CCTGGGGTCAAGGAACTACGGTCACCGTCTCCTCA 646. PM 98-B4 HCDR1 artificial
aa DFFMA 647. PM 98-B4 HCDR2 artificial aa TIVSDGGSTYYRDSVKG 648.
PM 98-B4 HCDR3 artificial aa RGNSGYYVMDA 649. PM 98-B4 HL
artificial aa
EVQLLESGGGLVQPGGSLRLSCAASGFTFSDFFMAWIRQAPGKGLEWVSTIVSDGGSTYYRDSVK
GRFTISRDNSQSTLYLQMDSLTAEDTAVYYCAKRGNSGYYVMDAWGQGTTVTVSSGGGGSGGGGS
GGGGSDIQMTQSPSTLSASVGEKVTITCRASSSVTYMHWYQQKPGKSPKLLIYDTSKVASGVPSR
FSGSGSGTAYTLTISSLQTDDFATYYCQQWSRNSPYTFGQGTKLEIK 650. PM 98-B4 HL
artificial nt
GAGGTGCAGCTGCTCGAGTCTGGTGGAGGCTTAGTGCAGCCTGGAGGGTCCCTAAGACTCTCCTG
TGCAGCCTCAGGATTCACTTTCAGTGACTTTTTCATGGCCTGGATCCGCCAGGCTCCAGGGAAGG
GGCTGGAGTGGGTCTCAACCATTGTTTCTGATGGTGGTAGCACTTACTATCGCGACTCCGTGAAG
GGCCGTTTCACTATCTCCAGAGATAATTCACAAAGCACCCTGTACCTGCAAATGGACAGTCTGAC
GGCTGAGGACACGGCCGTTTATTACTGTGCAAAACGCGGCAATTCGGGGTACTATGTTATGGATG
CCTGGGGTCAAGGAACTACGGTCACCGTCTCCTCAGGTGGTGGTGGTTCTGGCGGCGGCGGCTCC
GGTGGTGGTGGTTCTGACATCCAGATGACTCAGTCTCCATCAACCCTGTCTGCATCTGTGGGGGA
GAAAGTCACCATCACCTGCCGTGCCAGCTCCAGTGTGACTTACATGCACTGGTACCAGCAGAAGC
CAGGCAAATCCCCTAAATTATTGATTTATGACACATCCAAAGTGGCTTCTGGAGTCCCAAGTCGC
TTCAGTGGCAGTGGGTCTGGGACCGCATATACTCTCACAATCAGCTCCCTGCAGACTGATGACTT
TGCCACTTATTACTGTCAGCAGTGGAGTAGGAACTCACCCTACACGTTTGGACAGGGGACCAAGC
TGGAAATCAAA 651. PM 98-B4 HL .times. I2C artificial aa
EVQLLESGGGLVQPGGSLRLSCAASGFTFSDFFMAWIRQAPGKGLEWVSTIVSDGGSTYYRDSVK
HL
GRFTISRDNSQSTLYLQMDSLTAEDTAVYYCAKRGNSGYYVMDAWGQGTTVTVSSGGGGSGGGGS
GGGGSDIQMTQSPSTLSASVGEKVTITCRASSSVTYMHWYQQKPGKSPKLLIYDTSKVASGVPSR
FSGSGSGTAYTLTISSLQTDDFATYYCQQWSRNSPYTFGQGTKLEIKSGGGGSEVQLVESGGGLV
QPGGSLKLSCAASGFTFNKYAMNWVRQAPGKGLEWVARIRSKYNNYATYYADSVKDRFTISRDDS
KNTAYLQMNNLKTEDTAVYYCVRHGNFGNSYISYWAYWGQGTLVTVSSGGGGSGGGGSGGGGSQT
VVTQEPSLTVSPGGTVTLTCGSSTGAVTSGNYPNWVQQKPGQAPRGLIGGTKFLAPGTPARFSGS
LLGGKAALTLSGVQPEDEAEYYCVLWYSNRWVFGGGTKLTVL 652. PM 98-B4 HL .times.
I2C artificial nt
GAGGTGCAGCTGCTCGAGTCTGGTGGAGGCTTAGTGCAGCCTGGAGGGTCCCTAAGACTCTCCTG
HL
TGCAGCCTCAGGATTCACTTTCAGTGACTTTTTCATGGCCTGGATCCGCCAGGCTCCAGGGAAGG
GGCTGGAGTGGGTCTCAACCATTGTTTCTGATGGTGGTAGCACTTACTATCGCGACTCCGTGAAG
GGCCGTTTCACTATCTCCAGAGATAATTCACAAAGCACCCTGTACCTGCAAATGGACAGTCTGAC
GGCTGAGGACACGGCCGTTTATTACTGTGCAAAACGCGGCAATTCGGGGTACTATGTTATGGATG
CCTGGGGTCAAGGAACTACGGTCACCGTCTCCTCAGGTGGTGGTGGTTCTGGCGGCGGCGGCTCC
GGTGGTGGTGGTTCTGACATCCAGATGACTCAGTCTCCATCAACCCTGTCTGCATCTGTGGGGGA
GAAAGTCACCATCACCTGCCGTGCCAGCTCCAGTGTGACTTACATGCACTGGTACCAGCAGAAGC
CAGGCAAATCCCCTAAATTATTGATTTATGACACATCCAAAGTGGCTTCTGGAGTCCCAAGTCGC
TTCAGTGGCAGTGGGTCTGGGACCGCATATACTCTCACAATCAGCTCCCTGCAGACTGATGACTT
TGCCACTTATTACTGTCAGCAGTGGAGTAGGAACTCACCCTACACGTTTGGACAGGGGACCAAGC
TGGAAATCAAATCCGGAGGTGGTGGATCCGAGGTGCAGCTGGTCGAGTCTGGAGGAGGATTGGTG
CAGCCTGGAGGGTCATTGAAACTCTCATGTGCAGCCTCTGGATTCACCTTCAATAAGTACGCCAT
GAACTGGGTCCGCCAGGCTCCAGGAAAGGGTTTGGAATGGGTTGCTCGCATAAGAAGTAAATATA
ATAATTATGCAACATATTATGCCGATTCAGTGAAAGACAGGTTCACCATCTCCAGAGATGATTCA
AAAAACACTGCCTATCTACAAATGAACAACTTGAAAACTGAGGACACTGCCGTGTACTACTGTGT
GAGACATGGGAACTTCGGTAATAGCTACATATCCTACTGGGCTTACTGGGGCCAAGGGACTCTGG
TCACCGTCTCCTCAGGTGGTGGTGGTTCTGGCGGCGGCGGCTCCGGTGGTGGTGGTTCTCAGACT
GTTGTGACTCAGGAACCTTCACTCACCGTATCACCTGGTGGAACAGTCACACTCACTTGTGGCTC
CTCGACTGGGGCTGTTACATCTGGCAACTACCCAAACTGGGTCCAACAAAAACCAGGTCAGGCAC
CCCGTGGTCTAATAGGTGGGACTAAGTTCCTCGCCCCCGGTACTCCTGCCAGATTCTCAGGCTCC
CTGCTTGGAGGCAAGGCTGCCCTCACCCTCTCAGGGGTACAGCCAGAGGATGAGGCAGAATATTA
CTGTGTTCTATGGTACAGCAACCGCTGGGTGTTCGGTGGAGGAACCAAACTGACTGTCCTA 653.
PM 86-C3 VL artificial aa
DIQMTQSPSTLAASVGDRVTITCRASSSVTYMHWYQQKPGKAPKLLIYDTSKVASGVPNRFSGSG
SGTEFTLTISSLQTDDSATYYCQQWSRNSPYTFGQGTKLEIK 654. PM 86-C3 VL
artificial nt
GACATCCAGATGACTCAGTCTCCATCAACCCTGGCTGCATCTGTGGGGGACAGAGTCACCATCAC
CTGCCGTGCCAGCTCCAGTGTGACTTACATGCACTGGTACCAGCAGAAGCCAGGCAAAGCCCCTA
AATTATTGATTTATGACACATCCAAAGTGGCTTCTGGAGTCCCAAATCGCTTCAGTGGCAGTGGG
TCTGGGACCGAATTTACTCTCACAATCAGCTCCCTGCAGACTGATGACTCTGCCACTTATTACTG
TCAGCAGTGGAGTAGGAACTCACCCTACACGTTTGGACAGGGGACCAAGCTGGAAATCAAA 655.
PM 86-C3 LCDR1 artificial aa RASSSVTYMH 656. PM 86-C3 LCDR2
artificial aa DTSKVAS 657. PM 86-C3 LCDR3 artificial aa QQWSRNSPYT
658. PM 86-C3 VH artificial aa
EVQLLESGGGLVQPGGSLKLSCAASGFTFSDFFMAWVRQAPGKGLEWVATIVSDGGSTYYRDSVK
GRFTISRDNAKNTLYLQMNSLTAEDTAIYYCAKRGNSGYYVMDAWGQGTTVTVSS 659. PM
86-C3 VH artificial nt
GAGGTGCAGCTGCTCGAGTCTGGTGGAGGCTTAGTGCAGCCTGGAGGGTCCCTAAAACTCTCCTG
TGCAGCCTCAGGATTCACTTTCAGTGACTTTTTCATGGCCTGGGTCCGCCAGGCTCCAGGGAAGG
GGCTGGAGTGGGTCGCAACCATTGTTTCTGATGGTGGTAGCACTTACTATCGTGACTCCGTGAAG
GGCCGTTTCACTATCTCCAGAGATAATGCAAAAAACACCCTGTACCTGCAAATGAACAGTCTGAC
GGCTGAGGACACGGCCATTTATTACTGTGCAAAACGCGGCAATTCGGGGTACTATGTTATGGATG
CCTGGGGTCAAGGAACTACGGTCACCGTCTCCTCA 660. PM 86-C3 HCDR1 artificial
aa DFFMA 661. PM 86-C3 HCDR2 artificial aa TIVSDGGSTYYRDSVKG 662.
PM 86-C3 HCDR3 artificial aa RGNSGYYVMDA 663. PM 86-C3 HL
artificial aa
EVQLLESGGGLVQPGGSLKLSCAASGFTFSDFFMAWVRQAPGKGLEWVATIVSDGGSTYYRDSVK
GRFTISRDNAKNTLYLQMNSLTAEDTAIYYCAKRGNSGYYVMDAWGQGTTVTVSSGGGGSGGGGS
GGGGSDIQMTQSPSTLAASVGDRVTITCRASSSVTYMHWYQQKPGKAPKLLIYDTSKVASGVPNR
FSGSGSGTEFTLTISSLQTDDSATYYCQQWSRNSPYTFGQGTKLEIK 664. PM 86-C3 HL
artificial nt
GAGGTGCAGCTGCTCGAGTCTGGTGGAGGCTTAGTGCAGCCTGGAGGGTCCCTAAAACTCTCCTG
TGCAGCCTCAGGATTCACTTTCAGTGACTTTTTCATGGCCTGGGTCCGCCAGGCTCCAGGGAAGG
GGCTGGAGTGGGTCGCAACCATTGTTTCTGATGGTGGTAGCACTTACTATCGTGACTCCGTGAAG
GGCCGTTTCACTATCTCCAGAGATAATGCAAAAAACACCCTGTACCTGCAAATGAACAGTCTGAC
GGCTGAGGACACGGCCATTTATTACTGTGCAAAACGCGGCAATTCGGGGTACTATGTTATGGATG
CCTGGGGTCAAGGAACTACGGTCACCGTCTCCTCAGGTGGTGGTGGTTCTGGCGGCGGCGGCTCC
GGTGGTGGTGGTTCTGACATCCAGATGACTCAGTCTCCATCAACCCTGGCTGCATCTGTGGGGGA
CAGAGTCACCATCACCTGCCGTGCCAGCTCCAGTGTGACTTACATGCACTGGTACCAGCAGAAGC
CAGGCAAAGCCCCTAAATTATTGATTTATGACACATCCAAAGTGGCTTCTGGAGTCCCAAATCGC
TTCAGTGGCAGTGGGTCTGGGACCGAATTTACTCTCACAATCAGCTCCCTGCAGACTGATGACTC
TGCCACTTATTACTGTCAGCAGTGGAGTAGGAACTCACCCTACACGTTTGGACAGGGGACCAAGC
TGGAAATCAAA 665. PM 86-C3 HL .times. I2C artificial aa
EVQLLESGGGLVQPGGSLKLSCAASGFTFSDFFMAWVRQAPGKGLEWVATIVSDGGSTYYRDSVK
HL
GRFTISRDNAKNTLYLQMNSLTAEDTAIYYCAKRGNSGYYVMDAWGQGTTVTVSSGGGGSGGGGS
GGGGSDIQMTQSPSTLAASVGDRVTITCRASSSVTYMHWYQQKPGKAPKLLIYDTSKVASGVPNR
FSGSGSGTEFTLTISSLQTDDSATYYCQQWSRNSPYTFGQGTKLEIKSGGGGSEVQLVESGGGLV
QPGGSLKLSCAASGFTFNKYAMNWVRQAPGKGLEWVARIRSKYNNYATYYADSVKDRFTISRDDS
KNTAYLQMNNLKTEDTAVYYCVRHGNFGNSYISYWAYWGQGTLVTVSSGGGGSGGGGSGGGGSQT
VVTQEPSLTVSPGGTVTLTCGSSTGAVTSGNYPNWVQQKPGQAPRGLIGGTKFLAPGTPARFSGS
LLGGKAALTLSGVQPEDEAEYYCVLWYSNRWVFGGGTKLTVL 666. PM 86-C3 HL .times.
I2C artificial nt
GAGGTGCAGCTGCTCGAGTCTGGTGGAGGCTTAGTGCAGCCTGGAGGGTCCCTAAAACTCTCCTG
HL
TGCAGCCTCAGGATTCACTTTCAGTGACTTTTTCATGGCCTGGGTCCGCCAGGCTCCAGGGAAGG
GGCTGGAGTGGGTCGCAACCATTGTTTCTGATGGTGGTAGCACTTACTATCGTGACTCCGTGAAG
GGCCGTTTCACTATCTCCAGAGATAATGCAAAAAACACCCTGTACCTGCAAATGAACAGTCTGAC
GGCTGAGGACACGGCCATTTATTACTGTGCAAAACGCGGCAATTCGGGGTACTATGTTATGGATG
CCTGGGGTCAAGGAACTACGGTCACCGTCTCCTCAGGTGGTGGTGGTTCTGGCGGCGGCGGCTCC
GGTGGTGGTGGTTCTGACATCCAGATGACTCAGTCTCCATCAACCCTGGCTGCATCTGTGGGGGA
CAGAGTCACCATCACCTGCCGTGCCAGCTCCAGTGTGACTTACATGCACTGGTACCAGCAGAAGC
CAGGCAAAGCCCCTAAATTATTGATTTATGACACATCCAAAGTGGCTTCTGGAGTCCCAAATCGC
TTCAGTGGCAGTGGGTCTGGGACCGAATTTACTCTCACAATCAGCTCCCTGCAGACTGATGACTC
TGCCACTTATTACTGTCAGCAGTGGAGTAGGAACTCACCCTACACGTTTGGACAGGGGACCAAGC
TGGAAATCAAATCCGGAGGTGGTGGATCCGAGGTGCAGCTGGTCGAGTCTGGAGGAGGATTGGTG
CAGCCTGGAGGGTCATTGAAACTCTCATGTGCAGCCTCTGGATTCACCTTCAATAAGTACGCCAT
GAACTGGGTCCGCCAGGCTCCAGGAAAGGGTTTGGAATGGGTTGCTCGCATAAGAAGTAAATATA
ATAATTATGCAACATATTATGCCGATTCAGTGAAAGACAGGTTCACCATCTCCAGAGATGATTCA
AAAAACACTGCCTATCTACAAATGAACAACTTGAAAACTGAGGACACTGCCGTGTACTACTGTGT
GAGACATGGGAACTTCGGTAATAGCTACATATCCTACTGGGCTTACTGGGGCCAAGGGACTCTGG
TCACCGTCTCCTCAGGTGGTGGTGGTTCTGGCGGCGGCGGCTCCGGTGGTGGTGGTTCTCAGACT
GTTGTGACTCAGGAACCTTCACTCACCGTATCACCTGGTGGAACAGTCACACTCACTTGTGGCTC
CTCGACTGGGGCTGTTACATCTGGCAACTACCCAAACTGGGTCCAACAAAAACCAGGTCAGGCAC
CCCGTGGTCTAATAGGTGGGACTAAGTTCCTCGCCCCCGGTACTCCTGCCAGATTCTCAGGCTCC
CTGCTTGGAGGCAAGGCTGCCCTCACCCTCTCAGGGGTACAGCCAGAGGATGAGGCAGAATATTA
CTGTGTTCTATGGTACAGCAACCGCTGGGTGTTCGGTGGAGGAACCAAACTGACTGTCCTA 667.
PM 86-E12 VL artificial aa
DIQMTQSPSTLSASAGDRVTITCRASSSVTYMHWYQQKPGTAPKLLIYDTSKVASGVPSRFSGSG
SGTEFTLTISSVQPEDIATYYCQQWSRNSPYTFGQGTKLEIK 668. PM 86-E12 VL
artificial nt
GACATCCAGATGACTCAGTCTCCATCAACCCTGTCTGCATCTGCGGGGGACAGAGTCACCATCAC
CTGCCGTGCCAGCTCCAGTGTGACTTACATGCACTGGTACCAGCAGAAGCCAGGCACAGCCCCTA
AATTATTGATTTATGACACATCCAAAGTGGCTTCTGGAGTCCCAAGTCGCTTCAGTGGCAGTGGG
TCTGGGACCGAATTTACTCTCACAATCAGCTCCGTGCAGCCTGAAGACATTGCCACTTATTACTG
TCAGCAGTGGAGTAGGAACTCACCCTACACGTTTGGACAGGGGACCAAGCTGGAAATCAAA 669.
PM 86-E12 LCDR1 artificial aa RASSSVTYMH 670. PM 86-E12 LCDR2
artificial aa DTSKVAS 671. PM 86-E12 LCDR3 artificial aa QQWSRNSPYT
672. PM 86-E12 VH artificial aa
EVQLLESGGGLVQPGGSLRLSCAASGFTFSDFFMAWVRQAPTKGLEWVSTIVSDGGSTYYRDSVK
GRFTISRDNAKNTLYLQMNSLRAEDTAVYYCARRGNSGYYVMDAWGQGTTVTVSS 673. PM
86-E12 VH artificial nt
GAGGTGCAGCTGCTCGAGTCTGGTGGAGGCTTAGTGCAGCCTGGAGGGTCCCTAAGACTCTCCTG
TGCAGCCTCAGGATTCACTTTCAGTGACTTTTTCATGGCCTGGGTCCGCCAGGCTCCAACGAAGG
GGCTGGAGTGGGTCTCAACCATTGTTTCTGATGGTGGTAGCACTTACTATCGCGACTCCGTGAAG
GGCCGTTTCACTATCTCCAGAGATAATGCAAAAAACACCCTGTACCTGCAAATGAACAGTCTGAG
GGCTGAGGACACGGCCGCACGTTACTGTGCAAGACGCGGCAATTCGGGGTACTATGTTATGGATG
CCTGGGGTCAAGGAACTACGGTCACCGTCTCCTCA 674. PM 86-E12 HCDR1 artificial
aa DFFMA 675. PM 86-E12 HCDR2 artificial aa TIVSDGGSTYYRDSVKG 676.
PM 86-E12 HCDR3 artificial aa RGNSGYYVMDA 677. PM 86-E12 HL
artificial aa
EVQLLESGGGLVQPGGSLRLSCAASGFTFSDFFMAWVRQAPTKGLEWVSTIVSDGGSTYYRDSVK
GRFTISRDNAKNTLYLQMNSLRAEDTAVYYCARRGNSGYYVMDAWGQGTTVTVSSGGGGSGGGGS
GGGGSDIQMTQSPSTLSASAGDRVTITCRASSSVTYMHWYQQKPGTAPKLLIYDTSKVASGVPSR
FSGSGSGTEFTLTISSVQPEDIATYYCQQWSRNSPYTFGQGTKLEIK 678. PM 86-E12 HL
artificial nt
GAGGTGCAGCTGCTCGAGTCTGGTGGAGGCTTAGTGCAGCCTGGAGGGTCCCTAAGACTCTCCTG
TGCAGCCTCAGGATTCACTTTCAGTGACTTTTTCATGGCCTGGGTCCGCCAGGCTCCAACGAAGG
GGCTGGAGTGGGTCTCAACCATTGTTTCTGATGGTGGTAGCACTTACTATCGCGACTCCGTGAAG
GGCCGTTTCACTATCTCCAGAGATAATGCAAAAAACACCCTGTACCTGCAAATGAACAGTCTGAG
GGCTGAGGACACGGCCGTTTATTACTGTGCAAGACGCGGCAATTCGGGGTACTATGTTATGGATG
CCTGGGGTCAAGGAACTACGGTCACCGTCTCCTCAGGTGGTGGTGGTTCTGGCGGCGGCGGCTCC
GGTGGTGGTGGTTCTGACATCCAGATGACTCAGTCTCCATCAACCCTGTCTGCATCTGCGGGGGA
CAGAGTCACCATCACCTGCCGTGCCAGCTCCAGTGTGACTTACATGCACTGGTACCAGCAGAAGC
CAGGCACAGCCCCTAAATTATTGATTTATGACACATCCAAAGTGGCTTCTGGAGTCCCAAGTCGC
TTCAGTGGCAGTGGGTCTGGGACCGAATTTACTCTCACAATCAGCTCCGTGCAGCCTGAAGACAT
TGCCACTTATTACTGTCAGCAGTGGAGTAGGAACTCACCCTACACGTTTGGACAGGGGACCAAGC
TGGAAATCAAA 679. PM 86-E12 HL .times. artificial aa
EVQLLESGGGLVQPGGSLRLSCAASGFTFSDFFMAWVRQAPTKGLEWVSTIVSDGGSTYYRDSVK
I2C HL
GRFTISRDNAKNTLYLQMNSLRAEDTAVYYCARRGNSGYYVMDAWGQGTTVTVSSGGGGSGGGG- S
GGGGSDIQMTQSPSTLSASAGDRVTITCRASSSVTYMHWYQQKPGTAPKLLIYDTSKVASGVPSR
FSGSGSGTEFTLTISSVQPEDIATYYCQQWSRNSPYTFGQGTKLEIKSGGGGSEVQLVESGGGLV
QPGGSLKLSCAASGFTFNKYAMNWVRQAPGKGLEWVARIRSKYNNYATYYADSVKDRFTISRDDS
KNTAYLQMNNLKTEDTAVYYCVRHGNFGNSYISYWAYWGQGTLVTVSSGGGGSGGGGSGGGGSQT
VVTQEPSLTVSPGGTVTLTCGSSTGAVTSGNYPNWVQQKPGQAPRGLIGGTKFLAPGTPARFSGS
LLGGKAALTLSGVQPEDEAEYYCVLWYSNRWVFGGGTKLTVL 680. PM 86-E12 HL
.times. artificial nt
GAGGTGCAGCTGCTCGAGTCTGGTGGAGGCTTAGTGCAGCCTGGAGGGTCCCTAAGACTCTCCTG
I2C HL
TGCAGCCTCAGGATTCACTTTCAGTGACTTTTTCATGGCCTGGGTCCGCCAGGCTCCAACGAAG- G
GGCTGGAGTGGGTCTCAACCATTGTTTCTGATGGTGGTAGCACTTACTATCGCGACTCCGTGAAG
GGCCGTTTCACTATCTCCAGAGATAATGCAAAAAACACCCTGTACCTGCAAATGAACAGTCTGAG
GGCTGAGGACACGGCCGTTTATTACTGTGCAAGACGCGGCAATTCGGGGTACTATGTTATGGATG
CCTGGGGTCAAGGAACTACGGTCACCGTCTCCTCAGGTGGTGGTGGTTCTGGCGGCGGCGGCTCC
GGTGGTGGTGGTTCTGACATCCAGATGACTCAGTCTCCATCAACCCTGTCTGCATCTGCGGGGGA
CAGAGTCACCATCACCTGCCGTGCCAGCTCCAGTGTGACTTACATGCACTGGTACCAGCAGAAGC
CAGGCACAGCCCCTAAATTATTGATTTATGACACATCCAAAGTGGCTTCTGGAGTCCCAAGTCGC
TTCAGTGGCAGTGGGTCTGGGACCGAATTTACTCTCACAATCAGCTCCGTGCAGCCTGAAGACAT
TGCCACTTATTACTGTCAGCAGTGGAGTAGGAACTCACCCTACACGTTTGGACAGGGGACCAAGC
TGGAAATCAAATCCGGAGGTGGTGGATCCGAGGTGCAGCTGGTCGAGTCTGGAGGAGGATTGGTG
CAGCCTGGAGGGTCATTGAAACTCTCATGTGCAGCCTCTGGATTCACCTTCAATAAGTACGCCAT
GAACTGGGTCCGCCAGGCTCCAGGAAAGGGTTTGGAATGGGTTGCTCGCATAAGAAGTAAATATA
ATAATTATGCAACATATTATGCCGATTCAGTGAAAGACAGGTTCACCATCTCCAGAGATGATTCA
AAAAACACTGCCTATCTACAAATGAACAACTTGAAAACTGAGGACACTGCCGTGTACTACTGTGT
GAGACATGGGAACTTCGGTAATAGCTACATATCCTACTGGGCTTACTGGGGCCAAGGGACTCTGG
TCACCGTCTCCTCAGGTGGTGGTGGTTCTGGCGGCGGCGGCTCCGGTGGTGGTGGTTCTCAGACT
GTTGTGACTCAGGAACCTTCACTCACCGTATCACCTGGTGGAACAGTCACACTCACTTGTGGCTC
CTCGACTGGGGCTGTTACATCTGGCAACTACCCAAACTGGGTCCAACAAAAACCAGGTCAGGCAC
CCCGTGGTCTAATAGGTGGGACTAAGTTCCTCGCCCCCGGTACTCCTGCCAGATTCTCAGGCTCC
CTGCTTGGAGGCAAGGCTGCCCTCACCCTCTCAGGGGTACAGCCAGAGGATGAGGCAGAATATTA
CTGTGTTCTATGGTACAGCAACCGCTGGGTGTTCGGTGGAGGAACCAAACTGACTGTCCTA 681.
PM F1-A10 VL artificial aa
DIQMTQSPSTLSASVGDRVTITCRASSSVTYMHWYQQKPGKAPKLLIYDTSKVASGVPSRFSGSG
SGTEFTLTISSLQPDDFATYYCQQWSRNSPYTFGQGTKLEIK 682. PM F1-A10 VL
artificial nt
GACATCCAGATGACTCAGTCTCCATCAACCCTGTCTGCATCTGTGGGGGACAGAGTCACCATCAC
CTGCCGGGCCAGCTCCAGTGTGACTTACATGCACTGGTACCAGCAGAAGCCAGGCAAAGCCCCTA
AATTATTGATTTATGACACATCCAAAGTGGCTTCTGGAGTCCCAAGTCGCTTCAGTGGCAGTGGG
TCTGGGACCGAATTTACTCTCACAATCAGCTCCCTGCAGCCTGATGACTTTGCCACTTATTACTG
TCAGCAGTGGAGTAGGAACTCACCCTACACGTTTGGGCAGGGGACCAAGCTGGAAATCAAA 683.
PM F1-A10 LCDR1 artificial aa RASSSVTYMH 684. PM F1-A10 LCDR2
artificial aa DTSKVAS 685. PM F1-A10 LCDR3 artificial aa QQWSRNSPYT
686. PM F1-A10 VH artificial aa
EVQLLESGGGLVQPGGSLRLSCAASGFTFSDFFMAWVRQAPGRGLEWVSTIVSDGGSTYYRDSVK
GRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARRGNSGYYVMDAWGQGTTVTVSS 687. PM
F1-A10 VH artificial nt
GAGGTGCAGCTGCTCGAGTCTGGTGGAGGCTTAGTGCAGCCTGGAGGGTCCCTAAGACTCTCCTG
TGCAGCCTCAGGATTCACTTTCAGTGACTTTTTCATGGCCTGGGTCCGCCAGGCTCCAGGGAGGG
GGCTGGAGTGGGTCTCAACCATTGTTTCTGATGGTGGTAGCACTTACTATCGCGACTCCGTGAAG
GGCCGTTTCACTATCTCCAGAGATAATTCAAAAAACACCCTGTACCTGCAAATGAACAGTCTGAG
GGCTGAGGACACGGCCGTTTATTACTGTGCAAGACGCGGCAATTCGGGGTACTATGTTATGGATG
CCTGGGGTCAAGGAACTACGGTCACCGTCTCCTCA 688. PM F1-A10 HCDR1 artificial
aa DFFMA 689. PM F1-A10 HCDR2 artificial aa TIVSDGGSTYYRDSVKG 690.
PM F1-A10 HCDR3 artificial aa RGNSGYYVMDA 691. PM F1-A10 HL
artificial aa
EVQLLESGGGLVQPGGSLRLSCAASGFTFSDFFMAWVRQAPGRGLEWVSTIVSDGGSTYYRDSVK
GRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARRGNSGYYVMDAWGQGTTVTVSSGGGGSGGGGS
GGGGSDIQMTQSPSTLSASVGDRVTITCRASSSVTYMHWYQQKPGKAPKLLIYDTSKVASGVPSR
FSGSGSGTEFTLTISSLQPDDFATYYCQQWSRNSPYTFGQGTKLEIK 692. PM F1-A10 HL
artificial nt
GAGGTGCAGCTGCTCGAGTCTGGTGGAGGCTTAGTGCAGCCTGGAGGGTCCCTAAGACTCTCCTG
TGCAGCCTCAGGATTCACTTTCAGTGACTTTTTCATGGCCTGGGTCCGCCAGGCTCCAGGGAGGG
GGCTGGAGTGGGTCTCAACCATTGTTTCTGATGGTGGTAGCACTTACTATCGCGACTCCGTGAAG
GGCCGTTTCACTATCTCCAGAGATAATTCAAAAAACACCCTGTACCTGCAAATGAACAGTCTGAG
GGCTGAGGACACGGCCGTTTATTACTGTGCAAGACGCGGCAATTCGGGGTACTATGTTATGGATG
CCTGGGGTCAAGGAACTACGGTCACCGTCTCCTCAGGTGGTGGTGGTTCTGGCGGCGGCGGCTCC
GGTGGTGGTGGTTCTGACATCCAGATGACTCAGTCTCCATCAACCCTGTCTGCATCTGTGGGGGA
CAGAGTCACCATCACCTGCCGGGCCAGCTCCAGTGTGACTTACATGCACTGGTACCAGCAGAAGC
CAGGCAAAGCCCCTAAATTATTGATTTATGACACATCCAAAGTGGCTTCTGGAGTCCCAAGTCGC
TTCAGTGGCAGTGGGTCTGGGACCGAATTTACTCTCACAATCAGCTCCCTGCAGCCTGATGACTT
TGCCACTTATTACTGTCAGCAGTGGAGTAGGAACTCACCCTACACGTTTGGGCAGGGGACCAAGC
TGGAAATCAAA 693. PM F1-A10 HL .times. artificial aa
EVQLLESGGGLVQPGGSLRLSCAASGFTFSDFFMAWVRQAPGRGLEWVSTIVSDGGSTYYRDSVK
I2C HL
GRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARRGNSGYYVMDAWGQGTTVTVSSGGGGSGGGG- S
GGGGSDIQMTQSPSTLSASVGDRVTITCRASSSVTYMHWYQQKPGKAPKLLIYDTSKVASGVPSR
FSGSGSGTEFTLTISSLQPDDFATYYCQQWSRNSPYTFGQGTKLEIKSGGGGSEVQLVESGGGLV
QPGGSLKLSCAASGFTFNKYAMNWVRQAPGKGLEWVARIRSKYNNYATYYADSVKDRFTISRDDS
KNTAYLQMNNLKTEDTAVYYCVRHGNFGNSYISYWAYWGQGTLVTVSSGGGGSGGGGSGGGGSQT
VVTQEPSLTVSPGGTVTLTCGSSTGAVTSGNYPNWVQQKPGQAPRGLIGGTKFLAPGTPARFSGS
LLGGKAALTLSGVQPEDEAEYYCVLWYSNRWVFGGGTKLTVL 694. PM F1-A10 HL
.times. artificial nt
GAGGTGCAGCTGCTCGAGTCTGGTGGAGGCTTAGTGCAGCCTGGAGGGTCCCTAAGACTCTCCTG
I2C HL
TGCAGCCTCAGGATTCACTTTCAGTGACTTTTTCATGGCCTGGGTCCGCCAGGCTCCAGGGAGG- G
GGCTGGAGTGGGTCTCAACCATTGTTTCTGATGGTGGTAGCACTTACTATCGCGACTCCGTGAAG
GGCCGTTTCACTATCTCCAGAGATAATTCAAAAAACACCCTGTACCTGCAAATGAACAGTCTGAG
GGCTGAGGACACGGCCGTTTATTACTGTGCAAGACGCGGCAATTCGGGGTACTATGTTATGGATG
CCTGGGGTCAAGGAACTACGGTCACCGTCTCCTCAGGTGGTGGTGGTTCTGGCGGCGGCGGCTCC
GGTGGTGGTGGTTCTGACATCCAGATGACTCAGTCTCCATCAACCCTGTCTGCATCTGTGGGGGA
CAGAGTCACCATCACCTGCCGGGCCAGCTCCAGTGTGACTTACATGCACTGGTACCAGCAGAAGC
CAGGCAAAGCCCCTAAATTATTGATTTATGACACATCCAAAGTGGCTTCTGGAGTCCCAAGTCGC
TTCAGTGGCAGTGGGTCTGGGACCGAATTTACTCTCACAATCAGCTCCCTGCAGCCTGATGACTT
TGCCACTTATTACTGTCAGCAGTGGAGTAGGAACTCACCCTACACGTTTGGGCAGGGGACCAAGC
TGGAAATCAAATCCGGAGGTGGTGGATCCGAGGTGCAGCTGGTCGAGTCTGGAGGAGGATTGGTG
CAGCCTGGAGGGTCATTGAAACTCTCATGTGCAGCCTCTGGATTCACCTTCAATAAGTACGCCAT
GAACTGGGTCCGCCAGGCTCCAGGAAAGGGTTTGGAATGGGTTGCTCGCATAAGAAGTAAATATA
ATAATTATGCAACATATTATGCCGATTCAGTGAAAGACAGGTTCACCATCTCCAGAGATGATTCA
AAAAACACTGCCTATCTACAAATGAACAACTTGAAAACTGAGGACACTGCCGTGTACTACTGTGT
GAGACATGGGAACTTCGGTAATAGCTACATATCCTACTGGGCTTACTGGGGCCAAGGGACTCTGG
TCACCGTCTCCTCAGGTGGTGGTGGTTCTGGCGGCGGCGGCTCCGGTGGTGGTGGTTCTCAGACT
GTTGTGACTCAGGAACCTTCACTCACCGTATCACCTGGTGGAACAGTCACACTCACTTGTGGCTC
CTCGACTGGGGCTGTTACATCTGGCAACTACCCAAACTGGGTCCAACAAAAACCAGGTCAGGCAC
CCCGTGGTCTAATAGGTGGGACTAAGTTCCTCGCCCCCGGTACTCCTGCCAGATTCTCAGGCTCC
CTGCTTGGAGGCAAGGCTGCCCTCACCCTCTCAGGGGTACAGCCAGAGGATGAGGCAGAATATTA
CTGTGTTCTATGGTACAGCAACCGCTGGGTGTTCGGTGGAGGAACCAAACTGACTGTCCTA 695.
PM 99-F1 VL artificial aa
DIQMTQSPSTLSASVGDRVTITCRASSSVTYMHWYQQKPGKSPKLLIYDTSKVASGVPSRFSGSG
SGTSFTLTISSLQPEDIATYYCQQWSRNSPYTFGQGTKLEIK 696. PM 99-F1 VL
artificial nt
GACATCCAGATGACTCAGTCTCCATCAACCCTGTCTGCATCTGTGGGGGACAGAGTCACCATCAC
CTGCCGTGCCAGCTCCAGTGTGACTTACATGCACTGGTACCAGCAGAAGCCAGGCAAATCCCCTA
AATTATTGATTTATGACACATCCAAAGTGGCTTCTGGAGTCCCAAGTCGCTTCAGTGGCAGTGGG
TCTGGGACCTCATTTACTCTCACAATCAGCTCCCTGCAGCCTGAAGACATTGCCACTTATTACTG
TCAGCAGTGGAGTAGGAACTCACCCTACACGTTTGGACAGGGGACCAAGCTGGAAATCAAA 697.
PM 99-F1 LCDR1 artificial aa RASSSVTYMH 698. PM 99-F1 LCDR2
artificial aa DTSKVAS 699. PM 99-F1 LCDR3 artificial aa QQWSRNSPYT
700. PM 99-F1 VH artificial aa
EVQLLESGGGLVQPGGSLRLSCAASGFTFSDFFMAWVRQAPGRGLEWVSTIVSDGGSTYYRDSVK
GRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARRGNSGYYVMDAWGQGTTVTVSS 701. PM
99-F1 VH artificial nt
GAGGTGCAGCTGCTCGAGTCTGGTGGAGGCTTAGTGCAGCCTGGAGGGTCCCTAAGACTCTCCTG
TGCAGCCTCAGGATTCACTTTCAGTGACTTTTTCATGGCCTGGGTCCGCCAGGCTCCAGGGAGGG
GGCTGGAGTGGGTCTCAACCATTGTTTCTGATGGTGGTAGCACTTACTATCGCGACTCCGTGAAG
GGCCGTTTCACTATCTCCAGAGATAATTCAAAAAACACCCTGTACCTGCAAATGAACAGTCTGAG
GGCTGAGGACACGGCCGTTTATTACTGTGCAAGACGCGGCAATTCGGGGTACTATGTTATGGATG
CCTGGGGTCAAGGAACTACGGTCACCGTCTCCTCA 702. PM 99-F1 HCDR1 artificial
aa DFFMA 703. PM 99-F1 HCDR2 artificial aa TIVSDGGSTYYRDSVKG 704.
PM 99-F1 HCDR3 artificial aa RGNSGYYVMDA 705. PM 99-F1 HL
artificial aa
EVQLLESGGGLVQPGGSLRLSCAASGFTFSDFFMAWVRQAPGRGLEWVSTIVSDGGSTYYRDSVK
GRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARRGNSGYYVMDAWGQGTTVTVSSGGGGSGGGGS
GGGGSDIQMTQSPSTLSASVGDRVTITCRASSSVTYMHWYQQKPGKSPKLLTYDTSKVASGVPSR
FSGSGSGTSFTLTISSLQPEDIATYYCQQWSRNSPYTFGQGTKLEIK 706. PM 99-F1 HL
artificial nt
GAGGTGCAGCTGCTCGAGTCTGGTGGAGGCTTAGTGCAGCCTGGAGGGTCCCTAAGACTCTCCTG
TGCAGCCTCAGGATTCACTTTCAGTGACTTTTTCATGGCCTGGGTCCGCCAGGCTCCAGGGAGGG
GGCTGGAGTGGGTCTCAACCATTGTTTCTGATGGTGGTAGCACTTACTATCGCGACTCCGTGAAG
GGCCGTTTCACTATCTCCAGAGATAATTCAAAAAACACCCTGTACCTGCAAATGAACAGTCTGAG
GGCTGAGGACACGGCCGTTTATTACTGTGCAAGACGCGGCAATTCGGGGTACTATGTTATGGATG
CCTGGGGTCAAGGAACTACGGTCACCGTCTCCTCAGGTGGTGGTGGTTCTGGCGGCGGCGGCTCC
GGTGGTGGCGGTTCTGACATCCAGATGACTCAGTCTCCATCAACCCTGTCTGCATCTGTGGGGGA
CAGAGTCACCATCACCTGCCGTGCCAGCTCCAGTGTGACTTACATGCACTGGTACCAGCAGAAGC
CAGGCAAATCCCCTAAATTATTGATTTATGACACATCCAAAGTGGCTTCTGGAGTCCCAAGTCGC
TTCAGTGGCAGTGGGTCTGGGACCTCATTTACTCTCACAATCAGCTCCCTGCAGCCTGAAGACAT
TGCCACTTATTACTGTCAGCAGTGGAGTAGGAACTCACCCTACACGTTTGGACAGGGGACCAAGC
TGGAAATCAAA 707. PM 99-F1 HL .times. I2C artificial aa
EVQLLESGGGLVQPGGSLRLSCAASGFTFSDFFMAWVRQAPGRGLEWVSTIVSDGGSTYYRDSVK
HL
GRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARRGNSGYYVMDAWGQGTTVTVSSGGGGSGGGGS
GGGGSDIQMTQSPSTLSASVGDRVTITCRASSSVTYMHWYQQKPGKSPKLLIYDTSKVASGVPSR
FSGSGSGTSFTLTISSLQPEDIATYYCQQWSRNSPYTFGQGTKLEIKSGGGGSEVQLVESGGGLV
QPGGSLKLSCAASGFTFNKYAMNWVRQAPGKGLEWVARIRSKYNNYATYYADSVKDRFTISRDDS
KNTAYLQMNNLKTEDTAVYYCVRHGNFGNSYISYWAYWGQGTLVTVSSGGGGSGGGGSGGGGSQT
VVTQEPSLTVSPGGTVTLTCGSSTGAVTSGNYPNWVQQKPGQAPRGLIGGTKFLAPGTPARFSGS
LLGGKAALTLSGVQPEDEAEYYCVLWYSNRWVFGGGTKLTVL 708. PM 99-F1 HL .times.
I2C artificial nt
GAGGTGCAGCTGCTCGAGTCTGGTGGAGGCTTAGTGCAGCCTGGAGGGTCCCTAAGACTCTCCTG
HL
TGCAGCCTCAGGATTCACTTTCAGTGACTTTTTCATGGCCTGGGTCCGCCAGGCTCCAGGGAGGG
GGCTGGAGTGGGTCTCAACCATTGTTTCTGATGGTGGTAGCACTTACTATCGCGACTCCGTGAAG
GGCCGTTTCACTATCTCCAGAGATAATTCAAAAAACACCCTGTACCTGCAAATGAACAGTCTGAG
GGCTGAGGACACGGCCGTTTATTACTGTGCAAGACGCGGCAATTCGGGGTACTATGTTATGGATG
CCTGGGGTCAAGGAACTACGGTCACCGTCTCCTCAGGTGGTGGTGGTTCTGGCGGCGGCGGCTCC
GGTGGTGGCGGTTCTGACATCCAGATGACTCAGTCTCCATCAACCCTGTCTGCATCTGTGGGGGA
CAGAGTCACCATCACCTGCCGTGCCAGCTCCAGTGTGACTTACATGCACTGGTACCAGCAGAAGC
CAGGCAAATCCCCTAAATTATTGATTTATGACACATCCAAAGTGGCTTCTGGAGTCCCAAGTCGC
TTCAGTGGCAGTGGGTCTGGGACCTCATTTACTCTCACAATCAGCTCCCTGCAGCCTGAAGACAT
TGCCACTTATTACTGTCAGCAGTGGAGTAGGAACTCACCCTACACGTTTGGACAGGGGACCAAGC
TGGAAATCAAATCCGGAGGTGGTGGATCCGAGGTGCAGCTGGTCGAGTCTGGAGGAGGATTGGTG
CAGCCTGGAGGGTCATTGAAACTCTCATGTGCAGCCTCTGGATTCACCTTCAATAAGTACGCCAT
GAACTGGGTCCGCCAGGCTCCAGGAAAGGGTTTGGAATGGGTTGCTCGCATAAGAAGTAAATATA
ATAATTATGCAACATATTATGCCGATTCAGTGAAAGACAGGTTCACCATCTCCAGAGATGATTCA
AAAAACACTGCCTATCTACAAATGAACAACTTGAAAACTGAGGACACTGCCGTGTACTACTGTGT
GAGACATGGGAACTTCGGTAATAGCTACATATCCTACTGGGCTTACTGGGGCCAAGGGACTCTGG
TCACCGTCTCCTCAGGTGGTGGTGGTTCTGGCGGCGGCGGCTCCGGTGGTGGTGGTTCTCAGACT
GTTGTGACTCAGGAACCTTCACTCACCGTATCACCTGGTGGAACAGTCACACTCACTTGTGGCTC
CTCGACTGGGGCTGTTACATCTGGCAACTACCCAAACTGGGTCCAACAAAAACCAGGTCAGGCAC
CCCGTGGTCTAATAGGTGGGACTAAGTTCCTCGCCCCCGGTACTCCTGCCAGATTCTCAGGCTCC
CTGCTTGGAGGCAAGGCTGCCCTCACCCTCTCAGGGGTACAGCCAGAGGATGAGGCAGAATATTA
CTGTGTTCTATGGTACAGCAACCGCTGGGTGTTCGGTGGAGGAACCAAACTGACTGTCCTA 709.
PM 99-F5 VL artificial aa
DIQMTQSPSTLSASVGDRVTITCRASSSVTYMHWYQQKPGKSPKLLIYDTSKVASGVPSRFSGSG
SGTEFTLTISSLQPDDFATYYCQQWSRNSPYTFGQGTKLEIK 710. PM 99-F5 VL
artificial nt
GACATCCAGATGACTCAGTCTCCATCAACCCTGTCTGCATCTGTGGGGGACAGAGTCACCATCAC
CTGCCGTGCCAGCTCCAGTGTGACTTACATGCACTGGTACCAGCAGAAGCCAGGCAAATCCCCTA
AATTATTGATTTATGACACATCCAAAGTGGCTTCTGGAGTCCCAAGTCGCTTCAGTGGCAGTGGG
TCTGGGACCGAATTTACTCTCACAATCAGCTCCCTGCAGCCTGATGACTTTGCCACTTATTACTG
TCAGCAGTGGAGTAGGAACTCACCCTACACGTTTGGACAGGGGACCAAGCTGGAAATCAAA 711.
PM 99-F5 LCDR1 artificial aa RASSSVTYMH 712. PM 99-F5 LCDR2
artificial aa DTSKVAS 713. PM 99-F5 LCDR3 artificial aa QQWSRNSPYT
714. PM 99-F5 VH artificial aa
EVQLLESGGGLVQPGGSLKLSCAASGFTFSDFFMAWVRQAPGKGLEWVSTIVSDGGSTYYRDSVK
GRFTISRDNSKNTLYLQMNSLTAEDTAIYYCAKRGNSGYYVMDAWGQGTTVTVSS 715. PM
99-F5 VH artificial nt
GAGGTGCAGCTGCTCGAGTCTGGTGGAGGCTTAGTGCAGCCTGGAGGGTCCCTAAAACTCTCCTG
TGCAGCCTCAGGATTCACTTTCAGTGACTTTTTCATGGCCTGGGTCCGCCAGGCTCCAGGGAAGG
GGCTGGAGTGGGTCTCAACCATTGTTTCTGATGGTGGTAGCACTTACTATCGCGACTCCGTGAAG
GGCCGTTTCACTATCTCCAGAGATAATTCAAAAAACACCCTGTACCTGCAAATGAACAGTCTGAC
GGCTGAGGACACGGCCATTTATTACTGTGCAAAACGCGGCAATTCGGGGTACTATGTTATGGATG
CCTGGGGTCAAGGAACTACGGTCACCGTCTCCTCA 716. PM 99-F5 HCDR1 artificial
aa DFFMA 717. PM 99-F5 HCDR2 artificial aa TIVSDGGSTYYRDSVKG 718.
PM 99-F5 HCDR3 artificial aa RGNSGYYVMDA 719. PM 99-F5 HL
artificial aa
EVQLLESGGGLVQPGGSLKLSCAASGFTFSDFFMAWVRQAPGKGLEWVSTIVSDGGSTYYRDSVK
GRFTISRDNSKNTLYLQMNSLTAEDTAIYYCAKRGNSGYYVMDAWGQGTTVTVSSGGGGSGGGGS
GGGGSDIQMTQSPSTLSASVGDRVTITCRASSSVTYMHWYQQKPGKSPKLLIYDTSKVASGVPSR
FSGSGSGTEFTLTISSLQPDDFATYYCQQWSRNSPYTFGQGTKLEIK 720. PM 99-F5 HL
artificial nt
GAGGTGCAGCTGCTCGAGTCTGGTGGAGGCTTAGTGCAGCCTGGAGGGTCCCTAAAACTCTCCTG
TGCAGCCTCAGGATTCACTTTCAGTGACTTTTTCATGGCCTGGGTCCGCCAGGCTCCAGGGAAGG
GGCTGGAGTGGGTCTCAACCATTGTTTCTGATGGTGGTAGCACTTACTATCGCGACTCCGTGAAG
GGCCGTTTCACTATCTCCAGAGATAATTCAAAAAACACCCTGTACCTGCAAATGAACAGTCTGAC
GGCTGAGGACACGGCCATTTATTACTGTGCAAAACGCGGCAATTCGGGGTACTATGTTATGGATG
CCTGGGGTCAAGGAACTACGGTCACCGTCTCCTCAGGTGGTGGTGGTTCTGGCGGCGGCGGCTCC
GGTGGTGGTGGTTCTGACATCCAGATGACTCAGTCTCCATCAACCCTGTCTGCATCTGTGGGGGA
CAGAGTCACCATCACCTGCCGTGCCAGCTCCAGTGTGACTTACATGCACTGGTACCAGCAGAAGC
CAGGCAAATCCCCTAAATTATTGATTTATGACACATCCAAAGTGGCTTCTGGAGTCCCAAGTCGC
TTCAGTGGCAGTGGGTCTGGGACCGAATTTACTCTCACAATCAGCTCCCTGCAGCCTGATGACTT
TGCCACTTATTACTGTCAGCAGTGGAGTAGGAACTCACCCTACACGTTTGGACAGGGGACCAAGC
TGGAAATCAAA 721. PM 99-F5 HL .times. I2C artificial aa
EVQLLESGGGLVQPGGSLKLSCAASGFTFSDFFMAWVRQAPGKGLEWVSTIVSDGGSTYYRDSVK
HL
GRFTISRDNSKNTLYLQMNSLTAEDTAIYYCAKRGNSGYYVMDAWGQGTTVTVSSGGGGSGGGGS
GGGGSDIQMTQSPSTLSASVGDRVTITCRASSSVTYMHWYQQKPGKSPKLLIYDTSKVASGVPSR
FSGSGSGTEFTLTISSLQPDDFATYYCQQWSRNSPYTFGQGTKLEIKSGGGGSEVQLVESGGGLV
QPGGSLKLSCAASGFTFNKYAMNWVRQAPGKGLEWVARIRSKYNNYATYYADSVKDRFTISRDDS
KNTAYLQMNNLKTEDTAVYYCVRHGNFGNSYISYWAYWGQGTLVTVSSGGGGSGGGGSGGGGSQT
VVTQEPSLTVSPGGTVTLTCGSSTGAVTSGNYPNWVQQKPGQAPRGLIGGTKFLAPGTPARFSGS
LLGGKAALTLSGVQPEDEAEYYCVLWYSNRWVFGGGTKLTVL 722. PM 86-F6 VL
artificial aa
DIQMTQSPSTLSASVGEKVTITCRASSSVTYMHWYQQKPGKAPKLLIYDTSKVASGVPSRFSGSG
SGTEFTLTISSLEPEDFATYYCQQWSRNSPYTFGQGTKLEIK 723. PM 86-F6 VL
artificial nt
GACATCCAGATGACTCAGTCTCCATCAACCCTGTCTGCATCTGTGGGGGAGAAAGTCACCATCAC
CTGCCGTGCCAGCTCCAGTGTGACTTACATGCACTGGTACCAGCAGAAGCCAGGCAAAGCCCCTA
AATTATTGATTTATGACACATCCAAAGTGGCTTCTGGAGTCCCAAGTCGCTTCAGTGGCAGTGGG
TCTGGGACCGAATTTACTCTCACAATCAGCTCCCTGGAGCCTGAAGACTTTGCCACTTATTACTG
TCAGCAGTGGAGTAGGAACTCACCCTACACGTTTGGACAGGGGACCAAGCTGGAAATCAAA 724.
PM 86-F6 LCDR1 artificial aa RASSSVTYMH 725. PM 86-F6 LCDR2
artificial aa DTSKVAS 726. PM 86-F6 LCDR3 artificial aa QQWSRNSPYT
727. PM 86-F6 VH artificial aa
EVQLLESGGGLVQPGGSLKLSCAASGFTFSDFFMAWVRQAPGKGLEWVATIVSDGGSTYYRDSVK
GRFTISRDNSKNTLYLQMDSLTSEDTAIYYCARRGNSGYYVMDAWGQGTTVTVSS 728. PM
86-F6 VH artificial nt
GAGGTGCAGCTGCTCGAGTCTGGTGGAGGCTTAGTGCAGCCTGGAGGGTCCCTAAAACTCTCCTG
TGCAGCCTCAGGATTCACTTTCAGTGACTTTTTCATGGCCTGGGTCCGCCAGGCTCCAGGGAAGG
GGCTGGAGTGGGTCGCAACCATTGTTTCTGATGGTGGTAGCACTTACTATCGCGACTCCGTGAAG
GGCCGTTTCACTATTTCCAGAGATAATTCAAAAAACACCCTGTACCTGCAAATGGACAGTCTGAC
GTCTGAGGACACGGCCATTTATTACTGTGCAAGACGCGGCAATTCGGGGTACTATGTTATGGATG
CCTGGGGTCAAGGAACTACGGTCACCGTCTCCTCA 729. PM 86-F6 HCDR1 artificial
aa DFFMA 730. PM 86-F6 HCDR2 artificial aa TIVSDGGSTYYRDSVKG 731.
PM 86-F6 HCDR3 artificial aa RGNSGYYVMDA 732. PM 86-F6 HL
artificial aa
EVQLLESGGGLVQPGGSLKLSCAASGFTFSDFFMAWVRQAPGKGLEWVATIVSDGGSTYYRDSVK
GRFTISRDNSKNTLYLQMDSLTSEDTAIYYCARRGNSGYYVMDAWGQGTTVTVSSGGGGSGGGGS
GGGGSDIQMTQSPSTLSASVGEKVTITCRASSSVTYMHWYQQKPGKAPKLLIYDTSKVASGVPSR
FSGSGSGTEFTLTISSLEPEDFATYYCQQWSRNSPYTFGQGTKLEIK 733. PM 86-F6 HL
artificial nt
GAGGTGCAGCTGCTCGAGTCTGGTGGAGGCTTAGTGCAGCCTGGAGGGTCCCTAAAACTCTCCTG
TGCAGCCTCAGGATTCACTTTCAGTGACTTTTTCATGGCCTGGGTCCGCCAGGCTCCAGGGAAGG
GGCTGGAGTGGGTCGCAACCATTGTTTCTGATGGTGGTAGCACTTACTATCGCGACTCCGTGAAG
GGCCGTTTCACTATTTCCAGAGATAATTCAAAAAACACCCTGTACCTGCAAATGGACAGTCTGAC
GTCTGAGGACACGGCCATTTATTACTGTGCAAGACGCGGCAATTCGGGGTACTATGTTATGGATG
CCTGGGGTCAAGGAACTACGGTCACCGTCTCCTCAGGTGGTGGTGGTTCTGGCGGCGGCGGCTCC
GGTGGTGGTGGTTCTGACATCCAGATGACTCAGTCTCCATCAACCCTGTCTGCATCTGTGGGGGA
GAAAGTCACCATCACCTGCCGTGCCAGCTCCAGTGTGACTTACATGCACTGGTACCAGCAGAAGC
CAGGCAAAGCCCCTAAATTATTGATTTATGACACATCCAAAGTGGCTTCTGGAGTCCCAAGTCGC
TTCAGTGGCAGTGGGTCTGGGACCGAATTTACTCTCACAATCAGCTCCCTGGAGCCTGAAGACTT
TGCCACTTATTACTGTCAGCAGTGGAGTAGGAACTCACCCTACACGTTTGGACAGGGGACCAAGC
TGGAAATCAAA 734. PM 86-F6 HL .times. I2C artificial aa
EVQLLESGGGLVQPGGSLKLSCAASGFTFSDFFMAWVRQAPGKGLEWVATIVSDGGSTYYRDSVK
HL
GRFTISRDNSKNTLYLQMDSLTSEDTAIYYCARRGNSGYYVMDAWGQGTTVTVSSGGGGSGGGGS
GGGGSDIQMTQSPSTLSASVGEKVTITCRASSSVTYMHWYQQKPGKAPKLLIYDTSKVASGVPSR
FSGSGSGTEFTLTISSLEPEDFATYYCQQWSRNSPYTFGQGTKLEIKSGGGGSEVQLVESGGGLV
QPGGSLKLSCAASGFTFNKYAMNWVRQAPGKGLEWVARIRSKYNNYATYYADSVKDRFTISRDDS
KNTAYLQMNNLKTEDTAVYYCVRHGNFGNSYISYWAYWGQGTLVTVSSGGGGSGGGGSGGGGSQT
VVTQEPSLTVSPGGTVTLTCGSSTGAVTSGNYPNWVQQKPGQAPRGLIGGTKFLAPGTPARFSGS
LLGGKAALTLSGVQPEDEAEYYCVLWYSNRWVFGGGTKLTVL 735. PM 86-F6 HL .times.
I2C artificial nt
GAGGTGCAGCTGCTCGAGTCTGGTGGAGGCTTAGTGCAGCCTGGAGGGTCCCTAAAACTCTCCTG
HL
TGCAGCCTCAGGATTCACTTTCAGTGACTTTTTCATGGCCTGGGTCCGCCAGGCTCCAGGGAAGG
GGCTGGAGTGGGTCGCAACCATTGTTTCTGATGGTGGTAGCACTTACTATCGCGACTCCGTGAAG
GGCCGTTTCACTATTTCCAGAGATAATTCAAAAAACACCCTGTACCTGCAAATGGACAGTCTGAC
GTCTGAGGACACGGCCATTTATTACTGTGCAAGACGCGGCAATTCGGGGTACTATGTTATGGATG
CCTGGGGTCAAGGAACTACGGTCACCGTCTCCTCAGGTGGTGGTGGTTCTGGCGGCGGCGGCTCC
GGTGGTGGTGGTTCTGACATCCAGATGACTCAGTCTCCATCAACCCTGTCTGCATCTGTGGGGGA
GAAAGTCACCATCACCTGCCGTGCCAGCTCCAGTGTGACTTACATGCACTGGTACCAGCAGAAGC
CAGGCAAAGCCCCTAAATTATTGATTTATGACACATCCAAAGTGGCTTCTGGAGTCCCAAGTCGC
TTCAGTGGCAGTGGGTCTGGGACCGAATTTACTCTCACAATCAGCTCCCTGGAGCCTGAAGACTT
TGCCACTTATTACTGTCAGCAGTGGAGTAGGAACTCACCCTACACGTTTGGACAGGGGACCAAGC
TGGAAATCAAATCCGGAGGTGGTGGATCCGAGGTGCAGCTGGTCGAGTCTGGAGGAGGATTGGTG
CAGCCTGGAGGGTCATTGAAACTCTCATGTGCAGCCTCTGGATTCACCTTCAATAAGTACGCCAT
GAACTGGGTCCGCCAGGCTCCAGGAAAGGGTTTGGAATGGGTTGCTCGCATAAGAAGTAAATATA
ATAATTATGCAACATATTATGCCGATTCAGTGAAAGACAGGTTCACCATCTCCAGAGATGATTCA
AAAAACACTGCCTATCTACAAATGAACAACTTGAAAACTGAGGACACTGCCGTGTACTACTGTGT
GAGACATGGGAACTTCGGTAATAGCTACATATCCTACTGGGCTTACTGGGGCCAAGGGACTCTGG
TCACCGTCTCCTCAGGTGGTGGTGGTTCTGGCGGCGGCGGCTCCGGTGGTGGTGGTTCTCAGACT
GTTGTGACTCAGGAACCTTCACTCACCGTATCACCTGGTGGAACAGTCACACTCACTTGTGGCTC
CTCGACTGGGGCTGTTACATCTGGCAACTACCCAAACTGGGTCCAACAAAAACCAGGTCAGGCAC
CCCGTGGTCTAATAGGTGGGACTAAGTTCCTCGCCCCCGGTACTCCTGCCAGATTCTCAGGCTCC
CTGCTTGGAGGCAAGGCTGCCCTCACCCTCTCAGGGGTACAGCCAGAGGATGAGGCAGAATATTA
CTGTGTTCTATGGTACAGCAACCGCTGGGTGTTCGGTGGAGGAACCAAACTGACTGTCCTA 736.
PM99-F5 HL-I2C HL artificial nt
GAGGTGCAGCTGCTCGAGTCTGGTGGAGGCTTAGTGCAGCCTGGAGGGTCCCTAAAACTCTCCTG
TGCAGCCTCAGGATTCACTTTCAGTGACTTTTTCATGGCCTGGGTCCGCCAGGCTCCAGGGAAGG
GGCTGGAGTGGGTCTCAACCATTGTTTCTGATGGTGGTAGCACTTACTATCGCGACTCCGTGAAG
GGCCGTTTCACTATCTCCAGAGATAATTCAAAAAACACCCTGTACCTGCAAATGAACAGTCTGAC
GGCTGAGGACACGGCCATTTATTACTGTGCAAAACGCGGCAATTCGGGGTACTATGTTATGGATG
CCTGGGGTCAAGGAACTACGGTCACCGTCTCCTCAGGTGGTGGTGGTTCTGGCGGCGGCGGCTCC
GGTGGTGGTGGTTCTGACATCCAGATGACTCAGTCTCCATCAACCCTGTCTGCATCTGTGGGGGA
CAGAGTCACCATCACCTGCCGTGCCAGCTCCAGTGTGACTTACATGCACTGGTACCAGCAGAAGC
CAGGCAAATCCCCTAAATTATTGATTTATGACACATCCAAAGTGGCTTCTGGAGTCCCAAGTCGC
TTCAGTGGCAGTGGGTCTGGGACCGAATTTACTCTCACAATCAGCTCCCTGCAGCCTGATGACTT
TGCCACTTATTACTGTCAGCAGTGGAGTAGGAACTCACCCTACACGTTTGGACAGGGGACCAAGC
TGGAAATCAAATCCGGAGGTGGTGGATCCGAGGTGCAGCTGGTCGAGTCTGGAGGAGGATTGGTG
CAGCCTGGAGGGTCATTGAAACTCTCATGTGCAGCCTCTGGATTCACCTTCAATAAGTACGCCAT
GAACTGGGTCCGCCAGGCTCCAGGAAAGGGTTTGGAATGGGTTGCTCGCATAAGAAGTAAATATA
ATAATTATGCAACATATTATGCCGATTCAGTGAAAGACAGGTTCACCATCTCCAGAGATGATTCA
AAAAACACTGCCTATCTACAAATGAACAACTTGAAAACTGAGGACACTGCCGTGTACTACTGTGT
GAGACATGGGAACTTCGGTAATAGCTACATATCCTACTGGGCTTACTGGGGCCAAGGGACTCTGG
TCACCGTCTCCTCAGGTGGTGGTGGTTCTGGCGGCGGCGGCTCCGGTGGTGGTGGTTCTCAGACT
GTTGTGACTCAGGAACCTTCACTCACCGTATCACCTGGTGGAACAGTCACACTCACTTGTGGCTC
CTCGACTGGGGCTGTTACATCTGGCAACTACCCAAACTGGGTCCAACAAAAACCAGGTCAGGCAC
CCCGTGGTCTAATAGGTGGGACTAAGTTCCTCGCCCCCGGTACTCCTGCCAGATTCTCAGGCTCC
CTGCTTGGAGGCAAGGCTGCCCTCACCCTCTCAGGGGTACAGCCAGAGGATGAGGCAGAATATTA
CTGTGTTCTATGGTACAGCAACCGCTGGGTGTTCGGTGGAGGAACCAAACTGACTGTCCTA 737.
5'PM3-VH-A-XhoI artificial nt CCG GAT CTC GAG TCT GGC GGC GGA CTG
GTG AAG CCT GGC GRG TCC CTG ARG CTG TCC TGT 738. 3'PM3-VH-B
artificial nt CCA GTA CAT GTA GTA GTC GGA GAA GGT GAA GCC GGA GGC
GRY ACA GGA CAG CYT CAG GGA 739. 5'PM3-VH-C artificial nt TAC TAC
ATG TAC TGG RTC CGC CAG RCC CCT GRG AAG SGG CTG GAA TGG GTG KCC ATC
ATC TCC GAC GGC (SEQ ID NO. 739) 740. 3'PM3-VH-D artificial nt GGC
GTT GTC CCG GGA GAT GGT GAA CCG GCC CTT GAT GAT GTC GGA GTA GTA GGT
GTA GTA GCC GCC GTC GGA GAT GAT 741. 5'PM3-VH-E artificial nt TCC
CGG GAC AAC GCC AAG AAC ARC CTG TAC CTG CAG ATG ARC TCC CTG ARG KCC
GAG GAC ACC GCC RTG TAC TAC TGC RCC CGG GGC 742. 3'PM3-VH-F-BstEII
artificial nt CGA TAC GGT GAC CAG GGT GCC CTG GCC CCA GTA ATC CAT
GGC GCC GTG TCT CAG CAG AGG GAA GCC CCG GGY GCA GTA GTA 743.
5'PM4-VH-A-XhoI artificial nt CTT GAT CTC GAG TCT GGC GCC GAA STG
RWG RAG CCT GGC GCC TCC GTG AAG STG TCC TGC AAG GCC TCC GGC TAC
744. 3'PM4-VH-B artificial nt CCA TTC CAG GCC CTG CYC AGG CSY CTG
CCG CAS CCA GTT GAT GTC GAA GTA GGT GAA GGT GTA GCC GGA GGC CTT
745. 5'PM4-VH-C artificial nt CAG GGC CTG GAA TGG ATS GGC GGC ATC
TCC CCT GGC GAC GGC AAC ACC AAC TAC AAC GAG AAC TTC AAG 746.
3'PM4-VH-D artificial nt AT GTA GGC GGT GGA GMT GGA CKT GTC TMT GGT
CAK TGT GRC CYT GCC CTT GAA GTT CTC GTT GTA 747. 5'PM4-VH-E
artificial nt C TCC ACC GCC TAC ATS SAG CTG TCC CGG CTG ASA TCT GAS
GAC ACC GCC GTG TAC TWC TGC GCC AGG GAC GGC 748. 3'PM4-VH-F-BstEII
artificial nt AGA CAC GGT CAC CGT GGT GCC CTG GCC CCA AGA GTC CAT
GGC GTA GTA AGG GAA GTT GCC GTC CCT GGC GCA 749. 5'PM8-VH-A-XhoI
artificial nt CTT GAT CTC GAG TCC GGC SCT GAG STG RWG AAG CCT GGC
GCC TCC GTG AAG RTG TCC TGC AAG GCC TCC GGC TAC 750. 3'PM8-VH-B
artificial nt CCA TTC CAG CMS CTG GCC GGG TKY CTG TYT CAC CCA GTG
CAT CAC GTA GCC GGT GAA GGT GTA GCC GGA GGC CTT GCA 751. 5'PM8-VH-C
artificial nt CCC GGC CAG SKG CTG GAA TGG ATS GGC TAC ATC AAC CCT
TAC AAC GAC GTG ACC CGG TAC AAC GGC AAG TTC AAG 752. 3'PM8-VH-D
artificial nt TTC CAT GTA GGC GGT GGA GGM GKA CKT GTC KCT GGT AAK
GGT GRC TYT GCC CTT GAA CTT GCC GTT GTA 753. 5'PM8-VH-E artificial
nt TCC ACC GCC TAC ATG GAA CTG TCC RGC CTG ASG TCT GAG GAC ACC GCC
GTG TAC TAC TGC GCC AGG GGC 754. 3'PM8-VH-F-BstEII artificial nt
CGA TAC GGT GAC CAG AGT GCC TCT GCC CCA GGA GTC GAA GTA GTA CCA GTT
CTC GCC CCT GGC GCA GTA GTA 755. 5'PM3-VL-A-SacI artificial nt CTT
GAT GAG CTC CAG ATG ACC CAG TCC CCC ARS TYC MTG TCC RCC TCC GTG GGC
GAC AGA GTG ACC 756. 3'PM3-VL-B artificial nt GCC GGG CTT CTG CTG
AWA CCA GGC CAC GTT GGT GTC CAC GTT CTG GGA GGC CTT GCA GGT GAY GGT
CAC TCT GTC GCC 757. 5'PM3-VL-C artificial nt CAG CAG AAG CCC GGC
MAG KCC CCT AAG KCC CTG ATC TAC TCC GCC TCC TAC CGG TAC TCT 758.
3'PM3-VL-D artificial nt CAG GGT GAA GTC GGT GCC GGA CYC GGA GCC
GGA GAA CCG GKM AGG CAC GYC AGA GTA CCG GTA GGA 759. 5'PM3-VL-E
artificial nt ACC GAC TTC ACC CTG ACC ATC TCC ARC STG CAG YCT GAG
GAC YTC GCC RMG TAC TWC TGC CAG CAG TAC GAC 760. 3'PM3-VL-F-
artificial nt CGA GTA ACT AGT CGT ACG CTT GAT TTC CAG CTT GGT CCC
TCC GCC GAA BsiWI/SpeI GGT GTA AGG GTA GGA GTC GTA CTG CTG GCA 761.
5'PM4-VL-A-SacI artificial nt CTT GAT GAG CTC GTG ATG ACC CAG TCC
CCC CTG TCC CTG CCT GTG AYC CTG GGC SAM CMG GCC TCC ATC TCC TGC CGG
762. 3'PM4-VL-B artificial nt AAA CCA GTG CAG GTA GGT ATT GCC GTT
GGA GTG CAC CAG GGA CTG GGA GGA CCG GCA GGA GAT GGA GGC 763.
5'PM4-VL-C artificial nt ACC TAC CTG CAC TGG TTT CWG CAG ARG CCT
GGC CAG TCC CCT ARG CKG CTG ATC TAC ACC GTG TCC AAC CGG 764.
3'PM4-VL-D artificial nt CAG GGT GAA GTC GGT GCC GGA GCC GGA GCC
AGA GAA CCT GTC AGG CAC GCC GGA GAA CCG GTT GGA CAC GGT 765.
5'PM4-VL-E artificial nt GGC ACC GAC TTC ACC CTG AAG ATC TCC CGG
GTG GAG GCC GAA GAT STG GGC GTG TAC TWT TGC TCC CAG TCC ACC 766.
3'PM4-VL-F- artificial nt ACT CAG ACT AGT CGT ACG CTT GAT TTC CAG
CTT GGT CCC TCC GCC GAA BsiWI/SpeI GGT AGG CAC GTG GGT GGA CTG GGA
GCA 767. 5'PM8-VL-A-SacI artificial nt CTT GAT GAG CTC GTG ATG ACC
CAG TCT CCA SYC TCC CTG SCT GTG ACT CTG GGC CAG CSG GCC TCC ATC TCT
TGC CGG 768. 3'PM8-VL-B artificial nt CCA GTG CAT GAA GGT GTT GTC
GTA GGA GTC GAT GGA CTC GGA GGC CCG GCA AGA GAT GGA GGC 769.
5'PM8-VL-C artificial nt ACC TTC ATG CAC TGG TWT CAG CAG ARG CCT
GGC CAG YCT CCT MRC CKG CTG ATC TWC CGG GCC TCT ATC CTG GAA 770.
3'PM8-VL-D artificial nt CAG GGT GAA GTC GGT GCC GGA GCC AGA GCC
GGA GAA CCG GKC AGG GAY GCC GGA TTC CAG GAT AGA GGC CCG 771.
5'PM8-VL-E artificial nt ACC GAC TTC ACC CTG AMA ATC TMC CST GTG
GAG GCC GAS GAC GTG GSC RYC TAC TAC TGC CAC CAG 772. 3'PM8-VL-F-
artificial nt ACT CAG ACT AGT CGT ACG CTT GAT TTC CAG CTT GGT CCC
TCC GCC GAA BsiWI/SpeI GGT GTA AGG GTC CTC GAT GGA CTG GTG GCA GTA
GTA 773. PM84D7-H artificial aa
QVQLVQSGAEVKKPGASVKLSCKASGYTFTYFDINWVRQAPEQGLEWMGGISPGDGN
TNYNENFKGRVTMTIDTSSSTAYIELSRLTSDDTAVYYCARDGNFPYYAMDSWGQGTT VTVSS
774. PM84D7-HCDR1 artificial aa YFDIN 775. PM84D7-HCDR2 artificial
aa GISPGDGNTNYNENFKG 776. PM84D7-HCDR3 artificial aa DGNFPYYAMDS
777. PM84D7-H artificial nt
CAGGTGCAGCTGGTCCAGTCTGGCGCCGAAGTGAAGAAGCCTGGCGCCTCCGTGA
AGCTGTCCTGCAAGGCCTCCGGCTACACCTTCACCTACTTCGACATCAACTGGGTG
CGGCAGGCGCCTGAGCAGGGCCTGGAATGGATGGGCGGCATCTCCCCTGGCGAC
GGCAACACCAACTACAACGAGAACTTCAAGGGCAGGGTCACAATGACCATAGACAC
GTCCAGCTCCACCGCCTACATCGAGCTGTCCCGGCTGACATCTGACGACACCGCC
GTGTACTACTGCGCCAGGGACGGCAACTTCCCTTACTACGCCATGGACTCTTGGGG
CCAGGGCACCACGGTGACCGTCTCCTCA 778. PM84D7-L artificial aa
DIVMTQSPLSLPVTLGQQASISCRSSQSLVHSNGNTYLHWFQQRPGQSPKLLIYTVSNR
FSGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCSQSTHVPTFGGGTKLEIK 779.
PM84D7-LCDR1 artificial aa RSSQSLVHSNGNTYLH 780. PM84D7-LCDR2
artificial aa TVSNRFS 781. PM84D7-LCDR3 artificial aa SQSTHVPT 782.
PM84D7-L artificial nt
GACATCGTGATGACCCAGTCCCCCCTGTCCCTGCCTGTGACCCTGGGCCAACAGG
CCTCCATCTCCTGCCGGTCCTCCCAGTCCCTGGTGCACTCCAACGGCAATACCTAC
CTGCACTGGTTTCAGCAGAGGCCTGGCCAGTCCCCTAAGCTGCTGATCTACACCGT
GTCCAACCGGTTCTCCGGCGTGCCTGACAGGTTCTCTGGCTCCGGCTCCGGCACC
GACTTCACCCTGAAGATCTCCCGGGTGGAGGCCGAAGATGTGGGCGTGTACTATT
GCTCCCAGTCCACCCACGTGCCTACCTTCGGCGGAGGGACCAAGCTGGAAATCAAG
783. PM84D7-HL artificial aa
QVQLVQSGAEVKKPGASVKLSCKASGYTFTYFDINWVRQAPEQGLEWMGGISPGDGN
TNYNENFKGRVTMTIDTSSSTAYIELSRLTSDDTAVYYCARDGNFPYYAMDSWGQGTT
VTVSSGGGGSGGGGSGGGGSDIVMTQSPLSLPVTLGQQASISCRSSQSLVHSNGNTY
LHWFQQRPGQSPKLLIYTVSNRFSGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCSQ
STHVPTFGGGTKLEIK 784. PM84D7-HL artificial nt
CAGGTGCAGCTGGTCCAGTCTGGCGCCGAAGTGAAGAAGCCTGGCGCCTCCGTGA
AGCTGTCCTGCAAGGCCTCCGGCTACACCTTCACCTACTTCGACATCAACTGGGTG
CGGCAGGCGCCTGAGCAGGGCCTGGAATGGATGGGCGGCATCTCCCCTGGCGAC
GGCAACACCAACTACAACGAGAACTTCAAGGGCAGGGTCACAATGACCATAGACAC
GTCCAGCTCCACCGCCTACATCGAGCTGTCCCGGCTGACATCTGACGACACCGCC
GTGTACTACTGCGCCAGGGACGGCAACTTCCCTTACTACGCCATGGACTCTTGGGG
CCAGGGCACCACGGTGACCGTCTCCTCAGGTGGTGGTGGTTCTGGCGGCGGCGG
CTCCGGTGGTGGTGGTTCTGACATCGTGATGACCCAGTCCCCCCTGTCCCTGCCTG
TGACCCTGGGCCAACAGGCCTCCATCTCCTGCCGGTCCTCCCAGTCCCTGGTGCA
CTCCAACGGCAATACCTACCTGCACTGGTTTCAGCAGAGGCCTGGCCAGTCCCCTA
AGCTGCTGATCTACACCGTGTCCAACCGGTTCTCCGGCGTGCCTGACAGGTTCTCT
GGCTCCGGCTCCGGCACCGACTTCACCCTGAAGATCTCCCGGGTGGAGGCCGAAG
ATGTGGGCGTGTACTATTGCTCCCAGTCCACCCACGTGCCTACCTTCGGCGGAGG
GACCAAGCTGGAAATCAAG 785. PM84D7 HL .times. I2C artificial aa
QVQLVQSGAEVKKPGASVKLSCKASGYTFTYFDINWVRQAPEQGLEWMGGISPGDGN HL
TNYNENFKGRVTMTIDTSSSTAYIELSRLTSDDTAVYYCARDGNFPYYAMDSWGQGTT
VTVSSGGGGSGGGGSGGGGSDIVMTQSPLSLPVTLGQQASISCRSSQSLVHSNGNTY
LHWFQQRPGQSPKLLIYTVSNRFSGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCSQ
STHVPTFGGGTKLEIKSGGGGSEVQLVESGGGLVQPGGSLKLSCAASGFTFNKYAMN
WVRQAPGKGLEWVARIRSKYNNYATYYADSVKDRFTISRDDSKNTAYLQMNNLKTEDT
AVYYCVRHGNFGNSYISYWAYWGQGTLVTVSSGGGGSGGGGSGGGGSQTVVTQEP
SLTVSPGGTVTLTCGSSTGAVTSGNYPNWVQQKPGQAPRGLIGGTKFLAPGTPARFSG
SLLGGKAALTLSGVQPEDEAEYYCVLWYSNRWVFGGGTKLTVL 786. PM84D7 HL .times.
I2C artificial nt
CAGGTGCAGCTGGTCCAGTCTGGCGCCGAAGTGAAGAAGCCTGGCGCCTCCGTGA HL
AGCTGTCCTGCAAGGCCTCCGGCTACACCTTCACCTACTTCGACATCAACTGGGTG
CGGCAGGCGCCTGAGCAGGGCCTGGAATGGATGGGCGGCATCTCCCCTGGCGAC
GGCAACACCAACTACAACGAGAACTTCAAGGGCAGGGTCACAATGACCATAGACAC
GTCCAGCTCCACCGCCTACATCGAGCTGTCCCGGCTGACATCTGACGACACCGCC
GTGTACTACTGCGCCAGGGACGGCAACTTCCCTTACTACGCCATGGACTCTTGGGG
CCAGGGCACCACGGTGACCGTCTCCTCAGGTGGTGGTGGTTCTGGCGGCGGCGG
CTCCGGTGGTGGTGGTTCTGACATCGTGATGACCCAGTCCCCCCTGTCCCTGCCTG
TGACCCTGGGCCAACAGGCCTCCATCTCCTGCCGGTCCTCCCAGTCCCTGGTGCA
CTCCAACGGCAATACCTACCTGCACTGGTTTCAGCAGAGGCCTGGCCAGTCCCCTA
AGCTGCTGATCTACACCGTGTCCAACCGGTTCTCCGGCGTGCCTGACAGGTTCTCT
GGCTCCGGCTCCGGCACCGACTTCACCCTGAAGATCTCCCGGGTGGAGGCCGAAG
ATGTGGGCGTGTACTATTGCTCCCAGTCCACCCACGTGCCTACCTTCGGCGGAGG
GACCAAGCTGGAAATCAAGTCCGGAGGTGGTGGATCCGAGGTGCAGCTGGTCGAG
TCTGGAGGAGGATTGGTGCAGCCTGGAGGGTCATTGAAACTCTCATGTGCAGCCTC
TGGATTCACCTTCAATAAGTACGCCATGAACTGGGTCCGCCAGGCTCCAGGAAAGG
GTTTGGAATGGGTTGCTCGCATAAGAAGTAAATATAATAATTATGCAACATATTATGC
CGATTCAGTGAAAGACAGGTTCACCATCTCCAGAGATGATTCAAAAAACACTGCCTA
TCTACAAATGAACAACTTGAAAACTGAGGACACTGCCGTGTACTACTGTGTGAGACA
TGGGAACTTCGGTAATAGCTACATATCCTACTGGGCTTACTGGGGCCAAGGGACTC
TGGTCACCGTCTCCTCAGGTGGTGGTGGTTCTGGCGGCGGCGGCTCCGGTGGTG
GTGGTTCTCAGACTGTTGTGACTCAGGAACCTTCACTCACCGTATCACCTGGTGGA
ACAGTCACACTCACTTGTGGCTCCTCGACTGGGGCTGTTACATCTGGCAACTACCC
AAACTGGGTCCAACAAAAACCAGGTCAGGCACCCCGTGGTCTAATAGGTGGGACTA
AGTTCCTCGCCCCCGGTACTCCTGCCAGATTCTCAGGCTCCCTGCTTGGAGGCAAG
GCTGCCCTCACCCTCTCAGGGGTACAGCCAGAGGATGAGGCAGAATATTACTGTGT
TCTATGGTACAGCAACCGCTGGGTGTTCGGTGGAGGAACCAAACTGACTGTCCTA 787.
PM76A9-H artificial aa
QVQLVESGGGLVKPGGSLRLSCAASGFTFSDYYMYWVRQAPGKGLEWVAIISDGGYY
TYYSDIIKGRFTISRDNAKNNLYLQMNSLRAEDTAVYYCARGFPLLRHGAMDYWGQGTL VTVSS
788. PM76A9-HCDR1 artificial aa DYYMY 789. PM76A9-HCDR2 artificial
aa IISDGGYYTYYSDIIKG 790. PM76A9-HCDR3 artificial aa GFPLLRHGAMDY
791. PM76A9-H artificial nt
CAGGTGCAGCTGGTCGAGTCTGGCGGCGGACTGGTGAAGCCTGGCGGGTCCCTG
AGGCTGTCCTGTGCCGCCTCCGGCTTCACCTTCTCCGACTACTACATGTACTGGGT
CCGCCAGGCCCCTGGGAAGGGGCTGGAATGGGTGGCCATCATCTCCGACGGCGG
CTACTACACCTACTACTCCGACATCATCAAGGGCCGGTTCACCATCTCCCGGGACA
ACGCCAAGAACAATCTGTACCTGCAGATGAACTCCCTGAGGGCCGAGGACACCGC
CGTGTACTACTGCGCCCGGGGCTTCCCTCTGCTGAGACACGGCGCCATGGATTAC
TGGGGCCAGGGCACCCTGGTCACCGTCTCCTCA 792. PM76A9-L artificial aa
DIQMTQSPSSLSASVGDRVTITCKASQNVDTNVAWYQQKPGQAPKSLIYSASYRYSDV
PSRFSGSASGTDFTLTISSVQSEDFATYYCQQYDSYPYTFGGGTKLEIK 793. PM76A9-LCDR1
artificial aa KASQNVDTNVA 794. PM76A9-LCDR2 artificial aa SASYRYS
795. PM76A9-LCDR3 artificial aa QQYDSYPYT 796. PM76A9-L artificial
nt GACATCCAGATGACCCAGTCCCCCAGCTCCCTGTCCGCCTCCGTGGGCGACAGAG
TGACCATCACCTGCAAGGCCTCCCAGAACGTGGACACCAACGTGGCCTGGTATCA
GCAGAAGCCCGGCCAGGCCCCTAAGTCCCTGATCTACTCCGCCTCCTACCGGTAC
TCTGACGTGCCTTCCCGGTTCTCCGGCTCCGCGTCCGGCACCGACTTCACCCTGA
CCATCTCCAGCGTGCAGTCTGAGGACTTCGCCACGTACTACTGCCAGCAGTACGAC
TCCTACCCTTACACCTTCGGCGGAGGGACCAAGCTGGAAATCAAG 797. PM76A9-HL
artificial aa
QVQLVESGGGLVKPGGSLRLSCAASGFTFSDYYMYWVRQAPGKGLEWVAIISDGGYY
TYYSDIIKGRFTISRDNAKNNLYLQMNSLRAEDTAVYYCARGFPLLRHGAMDYWGQGTL
VTVSSGGGGSGGGGSGGGGSDIQMTQSPSSLSASVGDRVTITCKASQNVDTNVAWY
QQKPGQAPKSLIYSASYRYSDVPSRFSGSASGTDFTLTISSVQSEDFATYYCQQYDSYP
YTFGGGTKLEIK 798. PM76A9-HL artificial nt
CAGGTGCAGCTGGTCGAGTCTGGCGGCGGACTGGTGAAGCCTGGCGGGTCCCTG
AGGCTGTCCTGTGCCGCCTCCGGCTTCACCTTCTCCGACTACTACATGTACTGGGT
CCGCCAGGCCCCTGGGAAGGGGCTGGAATGGGTGGCCATCATCTCCGACGGCGG
CTACTACACCTACTACTCCGACATCATCAAGGGCCGGTTCACCATCTCCCGGGACA
ACGCCAAGAACAATCTGTACCTGCAGATGAACTCCCTGAGGGCCGAGGACACCGC
CGTGTACTACTGCGCCCGGGGCTTCCCTCTGCTGAGACACGGCGCCATGGATTAC
TGGGGCCAGGGCACCCTGGTCACCGTCTCCTCAGGTGGTGGTGGTTCTGGCGGC
GGCGGCTCCGGTGGTGGTGGTTCTGACATCCAGATGACCCAGTCCCCCAGCTCCC
TGTCCGCCTCCGTGGGCGACAGAGTGACCATCACCTGCAAGGCCTCCCAGAACGT
GGACACCAACGTGGCCTGGTATCAGCAGAAGCCCGGCCAGGCCCCTAAGTCCCTG
ATCTACTCCGCCTCCTACCGGTACTCTGACGTGCCTTCCCGGTTCTCCGGCTCCGC
GTCCGGCACCGACTTCACCCTGACCATCTCCAGCGTGCAGTCTGAGGACTTCGCCA
CGTACTACTGCCAGCAGTACGACTCCTACCCTTACACCTTCGGCGGAGGGACCAAG
CTGGAAATCAAG 799. PM76A9 HL .times. I2C artificial aa
QVQLVESGGGLVKPGGSLRLSCAASGFTFSDYYMYWVRQAPGKGLEWVAIISDGGYY HL
TYYSDIIKGRFTISRDNAKNNLYLQMNSLRAEDTAVYYCARGFPLLRHGAMDYWGQGTL
VTVSSGGGGSGGGGSGGGGSDIQMTQSPSSLSASVGDRVTITCKASQNVDTNVAWY
QQKPGQAPKSLIYSASYRYSDVPSRFSGSASGTDFTLTISSVQSEDFATYYCQQYDSYP
YTFGGGTKLEIKSGGGGSEVQLVESGGGLVQPGGSLKLSCAASGFTFNKYAMNWVRQ
APGKGLEWVARIRSKYNNYATYYADSVKDRFTISRDDSKNTAYLQMNNLKTEDTAVYY
CVRHGNFGNSYISYWAYWGQGTLVTVSSGGGGSGGGGSGGGGSQTVVTQEPSLTVS
PGGTVTLTCGSSTGAVTSGNYPNWVQQKPGQAPRGLIGGTKFLAPGTPARFSGSLLG
GKAALTLSGVQPEDEAEYYCVLWYSNRWVFGGGTKLTVL 800. PM76A9 HL .times. I2C
artificial nt
CAGGTGCAGCTGGTCGAGTCTGGCGGCGGACTGGTGAAGCCTGGCGGGTCCCTG HL
AGGCTGTCCTGTGCCGCCTCCGGCTTCACCTTCTCCGACTACTACATGTACTGGGT
CCGCCAGGCCCCTGGGAAGGGGCTGGAATGGGTGGCCATCATCTCCGACGGCGG
CTACTACACCTACTACTCCGACATCATCAAGGGCCGGTTCACCATCTCCCGGGACA
ACGCCAAGAACAATCTGTACCTGCAGATGAACTCCCTGAGGGCCGAGGACACCGC
CGTGTACTACTGCGCCCGGGGCTTCCCTCTGCTGAGACACGGCGCCATGGATTAC
TGGGGCCAGGGCACCCTGGTCACCGTCTCCTCAGGTGGTGGTGGTTCTGGCGGC
GGCGGCTCCGGTGGTGGTGGTTCTGACATCCAGATGACCCAGTCCCCCAGCTCCC
TGTCCGCCTCCGTGGGCGACAGAGTGACCATCACCTGCAAGGCCTCCCAGAACGT
GGACACCAACGTGGCCTGGTATCAGCAGAAGCCCGGCCAGGCCCCTAAGTCCCTG
ATCTACTCCGCCTCCTACCGGTACTCTGACGTGCCTTCCCGGTTCTCCGGCTCCGC
GTCCGGCACCGACTTCACCCTGACCATCTCCAGCGTGCAGTCTGAGGACTTCGCCA
CGTACTACTGCCAGCAGTACGACTCCTACCCTTACACCTTCGGCGGAGGGACCAAG
CTGGAAATCAAGTCCGGAGGTGGTGGATCCGAGGTGCAGCTGGTCGAGTCTGGAG
GAGGATTGGTGCAGCCTGGAGGGTCATTGAAACTCTCATGTGCAGCCTCTGGATTC
ACCTTCAATAAGTACGCCATGAACTGGGTCCGCCAGGCTCCAGGAAAGGGTTTGGA
ATGGGTTGCTCGCATAAGAAGTAAATATAATAATTATGCAACATATTATGCCGATTCA
GTGAAAGACAGGTTCACCATCTCCAGAGATGATTCAAAAAACACTGCCTATCTACAA
ATGAACAACTTGAAAACTGAGGACACTGCCGTGTACTACTGTGTGAGACATGGGAA
CTTCGGTAATAGCTACATATCCTACTGGGCTTACTGGGGCCAAGGGACTCTGGTCA
CCGTCTCCTCAGGTGGTGGTGGTTCTGGCGGCGGCGGCTCCGGTGGTGGTGGTTC
TCAGACTGTTGTGACTCAGGAACCTTCACTCACCGTATCACCTGGTGGAACAGTCA
CACTCACTTGTGGCTCCTCGACTGGGGCTGTTACATCTGGCAACTACCCAAACTGG
GTCCAACAAAAACCAGGTCAGGCACCCCGTGGTCTAATAGGTGGGACTAAGTTCCT
CGCCCCCGGTACTCCTGCCAGATTCTCAGGCTCCCTGCTTGGAGGCAAGGCTGCC
CTCACCCTCTCAGGGGTACAGCCAGAGGATGAGGCAGAATATTACTGTGTTCTATG
GTACAGCAACCGCTGGGTGTTCGGTGGAGGAACCAAACTGACTGTCCTA 801. PM76A9-H
artificial nt
CAGGTGCAGCTGGTCGAGTCTGGCGGCGGACTGGTCAAGCCTGGCGGCTCCCTG codon
optimized AGACTGTCTTGCGCTGCCTCCGGCTTCACCTTCTCCGACTACTACATGTACTGGGT
CCGCCAGGCTCCTGGCAAGGGACTGGAATGGGTGGCCATCATCTCCGACGGCGG
CTACTACACCTACTACTCCGACATCATCAAGGGCCGGTTCACCATCTCCAGGGACA
ACGCTAAGAACAACCTGTACCTGCAGATGAACTCCCTGAGGGCCGAGGACACCGC
CGTGTACTACTGCGCCAGGGGCTTCCCACTGCTGAGACACGGCGCCATGGATTAC
TGGGGCCAGGGCACCCTGGTCACAGTGTCCTCT 802. PM76A9-L artificial nt
GACATCCAGATGACCCAGTCCCCATCCTCCCTGTCTGCCTCCGTGGGCGACAGAGT codon
optimized GACCATCACATGCAAGGCCTCCCAGAACGTGGACACCAACGTGGCATGGTATCAG
CAGAAGCCAGGCCAGGCCCCTAAGTCCCTGATCTACTCTGCCTCCTACCGGTACTC
CGACGTGCCCTCCAGGTTCTCTGGCTCCGCCTCTGGCACCGACTTCACCCTGACCA
TCTCTTCCGTGCAGTCCGAGGACTTCGCTACCTACTACTGCCAGCAGTACGACTCC
TACCCTTACACCTTCGGCGGAGGCACCAAGCTGGAAATCAAG 803. PM76A9-HL
artificial nt
CAGGTGCAGCTGGTCGAGTCTGGCGGCGGACTGGTCAAGCCTGGCGGCTCCCTG codon
optimized AGACTGTCTTGCGCTGCCTCCGGCTTCACCTTCTCCGACTACTACATGTACTGGGT
CCGCCAGGCTCCTGGCAAGGGACTGGAATGGGTGGCCATCATCTCCGACGGCGG
CTACTACACCTACTACTCCGACATCATCAAGGGCCGGTTCACCATCTCCAGGGACA
ACGCTAAGAACAACCTGTACCTGCAGATGAACTCCCTGAGGGCCGAGGACACCGC
CGTGTACTACTGCGCCAGGGGCTTCCCACTGCTGAGACACGGCGCCATGGATTAC
TGGGGCCAGGGCACCCTGGTCACAGTGTCCTCTGGCGGAGGCGGAAGTGGAGGC
GGAGGAAGCGGAGGCGGCGGATCCGACATCCAGATGACCCAGTCCCCATCCTCCC
TGTCTGCCTCCGTGGGCGACAGAGTGACCATCACATGCAAGGCCTCCCAGAACGT
GGACACCAACGTGGCATGGTATCAGCAGAAGCCAGGCCAGGCCCCTAAGTCCCTG
ATCTACTCTGCCTCCTACCGGTACTCCGACGTGCCCTCCAGGTTCTCTGGCTCCGC
CTCTGGCACCGACTTCACCCTGACCATCTCTTCCGTGCAGTCCGAGGACTTCGCTA
CCTACTACTGCCAGCAGTACGACTCCTACCCTTACACCTTCGGCGGAGGCACCAAG
CTGGAAATCAAG 804. PM76A9 HL .times. I2C artificial nt
CAGGTGCAGCTGGTCGAGTCTGGCGGCGGACTGGTCAAGCCTGGCGGCTCCCTG HL_codon
optimized AGACTGTCTTGCGCTGCCTCCGGCTTCACCTTCTCCGACTACTACATGTACTGGGT
CCGCCAGGCTCCTGGCAAGGGACTGGAATGGGTGGCCATCATCTCCGACGGCGG
CTACTACACCTACTACTCCGACATCATCAAGGGCCGGTTCACCATCTCCAGGGACA
ACGCTAAGAACAACCTGTACCTGCAGATGAACTCCCTGAGGGCCGAGGACACCGC
CGTGTACTACTGCGCCAGGGGCTTCCCACTGCTGAGACACGGCGCCATGGATTAC
TGGGGCCAGGGCACCCTGGTCACAGTGTCCTCTGGCGGAGGCGGAAGTGGAGGC
GGAGGAAGCGGAGGCGGCGGATCCGACATCCAGATGACCCAGTCCCCATCCTCCC
TGTCTGCCTCCGTGGGCGACAGAGTGACCATCACATGCAAGGCCTCCCAGAACGT
GGACACCAACGTGGCATGGTATCAGCAGAAGCCAGGCCAGGCCCCTAAGTCCCTG
ATCTACTCTGCCTCCTACCGGTACTCCGACGTGCCCTCCAGGTTCTCTGGCTCCGC
CTCTGGCACCGACTTCACCCTGACCATCTCTTCCGTGCAGTCCGAGGACTTCGCTA
CCTACTACTGCCAGCAGTACGACTCCTACCCTTACACCTTCGGCGGAGGCACCAAG
CTGGAAATCAAGTCCGGCGGAGGGGGCTCTGAAGTGCAGCTGGTGGAAAGCGGA
GGGGGACTGGTGCAGCCCGGGGGAAGTCTGAAGCTGTCCTGTGCCGCCAGCGGC
TTTACCTTCAACAAGTACGCCATGAATTGGGTCCGACAGGCCCCAGGGAAAGGCCT
GGAATGGGTGGCACGGATCCGGTCCAAGTACAACAACTACGCCACCTACTACGCT
GACTCCGTGAAGGACAGATTCACCATCAGCCGGGACGACTCTAAGAACACCGCCTA
TCTGCAGATGAACAACCTGAAAACCGAGGATACAGCTGTGTACTATTGTGTGCGGC
ACGGCAACTTCGGCAACTCCTACATCTCCTACTGGGCCTATTGGGGACAGGGAACA
CTGGTCACCGTGTCTAGCGGAGGTGGCGGAAGTGGGGGAGGCGGATCTGGCGGT
GGCGGATCCCAGACCGTGGTCACCCAGGAACCTTCCCTGACCGTCTCCCCAGGCG
GCACCGTGACCCTGACCTGTGGCTCCTCTACCGGCGCTGTGACCTCCGGCAACTA
CCCTAACTGGGTGCAGCAGAAACCCGGACAGGCTCCTAGAGGCCTGATCGGCGGC
ACCAAGTTTCTGGCCCCTGGCACCCCTGCCAGATTCTCCGGCTCCCTGCTGGGAG
GCAAGGCCGCTCTGACCCTGTCTGGCGTGCAGCCTGAGGACGAGGCCGAGTACTA
CTGTGTGCTGTGGTACTCCAACAGATGGGTGTTCGGAGGCGGCACAAAGCTGACC GTGCTG 805.
PM76B10-H artificial aa
QVQLVESGGGLVKPGESLRLSCAASGFTFSDYYMYWVRQAPGKGLEWVAIISDGGYY
TYYSDIIKGRFTISRDNAKNSLYLQMNSLKAEDTAVYYCARGFPLLRHGAMDYWGQGTL VTVSS
806. PM76B10-HCDR1 artificial aa DYYMY 807. PM76B10-HCDR2
artificial aa IISDGGYYTYYSDIIKG 808. PM76B10-HCDR3 artificial aa
GFPLLRHGAMDY 809. PM76B10-H artificial nt
CAGGTGCAGCTGGTCGAGTCTGGCGGCGGACTGGTGAAGCCTGGCGAGTCCCTG
AGGCTGTCCTGTGCCGCCTCCGGCTTCACCTTCTCCGACTACTACATGTACTGGGT
CCGCCAGGCCCCTGGGAAGGGGCTGGAATGGGTGGCCATCATCTCCGACGGCGG
CTACTACACCTACTACTCCGACATCATCAAGGGCCGGTTCACCATCTCCCGGGACA
ACGCCAAGAACAGCCTGTACCTGCAGATGAACTCCCTGAAGGCCGAGGACACCGC
CGTGTACTACTGCGCCCGGGGCTTCCCTCTGCTGAGACACGGCGCCATGGATTAC
TGGGGCCAGGGCACCCTGGTCACCGTCTCCTCA
810. PM76B10-L artificial aa
DIQMTQSPSSLSASVGDRVTITCKASQNVDTNVAWYQQKPGQAPKSLIYSASYRYSDV
PSRFSGSASGTDFTLTISSVQSEDFATYYCQQYDSYPYTFGGGTKLEIK 811.
PM76B10-LCDR1 artificial aa KASQNVDTNVA 812. PM76B10-LCDR2
artificial aa SASYRYS 813. PM76B10-LCDR3 artificial aa QQYDSYPYT
814. PM76B10-L artificial nt
GACATCCAGATGACCCAGTCCCCCAGCTCCCTGTCCGCCTCCGTGGGCGACAGAG
TGACCATCACCTGCAAGGCCTCCCAGAACGTGGACACCAACGTGGCCTGGTATCA
GCAGAAGCCCGGCCAGGCCCCTAAGTCCCTGATCTACTCCGCCTCCTACCGGTAC
TCTGACGTGCCTTCCCGGTTCTCCGGCTCCGCGTCCGGCACCGACTTCACCCTGA
CCATCTCCAGCGTGCAGTCTGAGGACTTCGCCACGTACTACTGCCAGCAGTACGAC
TCCTACCCTTACACCTTCGGCGGAGGGACCAAGCTGGAAATCAAG 815. PM76B10-HL
artificial aa
QVQLVESGGGLVKPGESLRLSCAASGFTFSDYYMYWVRQAPGKGLEWVAIISDGGYY
TYYSDIIKGRFTISRDNAKNSLYLQMNSLKAEDTAVYYCARGFPLLRHGAMDYWGQGTL
VTVSSGGGGSGGGGSGGGGSDIQMTQSPSSLSASVGDRVTITCKASQNVDTNVAWY
QQKPGQAPKSLIYSASYRYSDVPSRFSGSASGTDFTLTISSVQSEDFATYYCQQYDSYP
YTFGGGTKLEIK 816. PM76B10-HL artificial nt
CAGGTGCAGCTGGTCGAGTCTGGCGGCGGACTGGTGAAGCCTGGCGAGTCCCTG
AGGCTGTCCTGTGCCGCCTCCGGCTTCACCTTCTCCGACTACTACATGTACTGGGT
CCGCCAGGCCCCTGGGAAGGGGCTGGAATGGGTGGCCATCATCTCCGACGGCGG
CTACTACACCTACTACTCCGACATCATCAAGGGCCGGTTCACCATCTCCCGGGACA
ACGCCAAGAACAGCCTGTACCTGCAGATGAACTCCCTGAAGGCCGAGGACACCGC
CGTGTACTACTGCGCCCGGGGCTTCCCTCTGCTGAGACACGGCGCCATGGATTAC
TGGGGCCAGGGCACCCTGGTCACCGTCTCCTCAGGTGGTGGTGGTTCTGGCGGC
GGCGGCTCCGGTGGTGGTGGTTCTGACATCCAGATGACCCAGTCCCCCAGCTCCC
TGTCCGCCTCCGTGGGCGACAGAGTGACCATCACCTGCAAGGCCTCCCAGAACGT
GGACACCAACGTGGCCTGGTATCAGCAGAAGCCCGGCCAGGCCCCTAAGTCCCTG
ATCTACTCCGCCTCCTACCGGTACTCTGACGTGCCTTCCCGGTTCTCCGGCTCCGC
GTCCGGCACCGACTTCACCCTGACCATCTCCAGCGTGCAGTCTGAGGACTTCGCCA
CGTACTACTGCCAGCAGTACGACTCCTACCCTTACACCTTCGGCGGAGGGACCAAG
CTGGAAATCAAG 817. PM76B10 HL .times. I2C artificial aa
QVQLVESGGGLVKPGESLRLSCAASGFTFSDYYMYWVRQAPGKGLEWVAIISDGGYY HL
TYYSDIIKGRFTISRDNAKNSLYLQMNSLKAEDTAVYYCARGFPLLRHGAMDYWGQGTL
VTVSSGGGGSGGGGSGGGGSDIQMTQSPSSLSASVGDRVTITCKASQNVDTNVAWY
QQKPGQAPKSLIYSASYRYSDVPSRFSGSASGTDFTLTISSVQSEDFATYYCQQYDSYP
YTFGGGTKLEIKSGGGGSEVQLVESGGGLVQPGGSLKLSCAASGFTFNKYAMNWVRQ
APGKGLEWVARIRSKYNNYATYYADSVKDRFTISRDDSKNTAYLQMNNLKTEDTAVYY
CVRHGNFGNSYISYWAYWGQGTLVTVSSGGGGSGGGGSGGGGSQTVVTQEPSLTVS
PGGTVTLTCGSSTGAVTSGNYPNWVQQKPGQAPRGLIGGTKFLAPGTPARFSGSLLG
GKAALTLSGVQPEDEAEYYCVLWYSNRWVFGGGTKLTVL 818. PM76B10 HL .times. I2C
artificial nt
CAGGTGCAGCTGGTCGAGTCTGGCGGCGGACTGGTGAAGCCTGGCGAGTCCCTG HL
AGGCTGTCCTGTGCCGCCTCCGGCTTCACCTTCTCCGACTACTACATGTACTGGGT
CCGCCAGGCCCCTGGGAAGGGGCTGGAATGGGTGGCCATCATCTCCGACGGCGG
CTACTACACCTACTACTCCGACATCATCAAGGGCCGGTTCACCATCTCCCGGGACA
ACGCCAAGAACAGCCTGTACCTGCAGATGAACTCCCTGAAGGCCGAGGACACCGC
CGTGTACTACTGCGCCCGGGGCTTCCCTCTGCTGAGACACGGCGCCATGGATTAC
TGGGGCCAGGGCACCCTGGTCACCGTCTCCTCAGGTGGTGGTGGTTCTGGCGGC
GGCGGCTCCGGTGGTGGTGGTTCTGACATCCAGATGACCCAGTCCCCCAGCTCCC
TGTCCGCCTCCGTGGGCGACAGAGTGACCATCACCTGCAAGGCCTCCCAGAACGT
GGACACCAACGTGGCCTGGTATCAGCAGAAGCCCGGCCAGGCCCCTAAGTCCCTG
ATCTACTCCGCCTCCTACCGGTACTCTGACGTGCCTTCCCGGTTCTCCGGCTCCGC
GTCCGGCACCGACTTCACCCTGACCATCTCCAGCGTGCAGTCTGAGGACTTCGCCA
CGTACTACTGCCAGCAGTACGACTCCTACCCTTACACCTTCGGCGGAGGGACCAAG
CTGGAAATCAAGTCCGGAGGTGGTGGATCCGAGGTGCAGCTGGTCGAGTCTGGAG
GAGGATTGGTGCAGCCTGGAGGGTCATTGAAACTCTCATGTGCAGCCTCTGGATTC
ACCTTCAATAAGTACGCCATGAACTGGGTCCGCCAGGCTCCAGGAAAGGGTTTGGA
ATGGGTTGCTCGCATAAGAAGTAAATATAATAATTATGCAACATATTATGCCGATTCA
GTGAAAGACAGGTTCACCATCTCCAGAGATGATTCAAAAAACACTGCCTATCTACAA
ATGAACAACTTGAAAACTGAGGACACTGCCGTGTACTACTGTGTGAGACATGGGAA
CTTCGGTAATAGCTACATATCCTACTGGGCTTACTGGGGCCAAGGGACTCTGGTCA
CCGTCTCCTCAGGTGGTGGTGGTTCTGGCGGCGGCGGCTCCGGTGGTGGTGGTTC
TCAGACTGTTGTGACTCAGGAACCTTCACTCACCGTATCACCTGGTGGAACAGTCA
CACTCACTTGTGGCTCCTCGACTGGGGCTGTTACATCTGGCAACTACCCAAACTGG
GTCCAACAAAAACCAGGTCAGGCACCCCGTGGTCTAATAGGTGGGACTAAGTTCCT
CGCCCCCGGTACTCCTGCCAGATTCTCAGGCTCCCTGCTTGGAGGCAAGGCTGCC
CTCACCCTCTCAGGGGTACAGCCAGAGGATGAGGCAGAATATTACTGTGTTCTATG
GTACAGCAACCGCTGGGTGTTCGGTGGAGGAACCAAACTGACTGTCCTA 819. PM76B10-H
artificial nt
CAGGTGCAGCTGGTCGAGTCTGGCGGCGGACTGGTCAAGCCTGGCGAGTCCCTGA codon
optimized GACTGTCTTGCGCTGCCTCCGGCTTCACCTTCTCCGACTACTACATGTACTGGGTC
CGCCAGGCTCCTGGCAAGGGACTGGAATGGGTGGCCATCATCTCCGACGGCGGCT
ACTACACCTACTACTCCGACATCATCAAGGGCCGGTTCACCATCTCCAGGGACAAC
GCCAAGAACTCCCTGTACCTGCAGATGAACTCCCTGAAGGCCGAGGACACCGCCG
TGTACTACTGCGCCAGGGGCTTCCCACTGCTGAGACACGGCGCCATGGATTACTG
GGGCCAGGGCACCCTGGTCACAGTGTCCTCT 820. PM76B10-L artificial nt
GACATCCAGATGACCCAGTCCCCATCCTCCCTGTCTGCCTCCGTGGGCGACAGAGT codon
optimized GACCATCACATGCAAGGCCTCCCAGAACGTGGACACCAACGTGGCATGGTATCAG
CAGAAGCCAGGCCAGGCCCCTAAGTCCCTGATCTACTCTGCCTCCTACCGGTACTC
CGACGTGCCCTCCAGGTTCTCTGGCTCCGCCTCTGGCACCGACTTCACCCTGACCA
TCTCTTCCGTGCAGTCCGAGGACTTCGCTACCTACTACTGCCAGCAGTACGACTCC
TACCCTTACACCTTCGGCGGAGGCACCAAGCTGGAAATCAAG 821. PM76B10-HL
artificial nt
CAGGTGCAGCTGGTCGAGTCTGGCGGCGGACTGGTCAAGCCTGGCGAGTCCCTGA codon
optimized GACTGTCTTGCGCTGCCTCCGGCTTCACCTTCTCCGACTACTACATGTACTGGGTC
CGCCAGGCTCCTGGCAAGGGACTGGAATGGGTGGCCATCATCTCCGACGGCGGCT
ACTACACCTACTACTCCGACATCATCAAGGGCCGGTTCACCATCTCCAGGGACAAC
GCCAAGAACTCCCTGTACCTGCAGATGAACTCCCTGAAGGCCGAGGACACCGCCG
TGTACTACTGCGCCAGGGGCTTCCCACTGCTGAGACACGGCGCCATGGATTACTG
GGGCCAGGGCACCCTGGTCACAGTGTCCTCTGGCGGAGGCGGAAGTGGAGGCGG
AGGAAGCGGAGGCGGCGGATCCGACATCCAGATGACCCAGTCCCCATCCTCCCTG
TCTGCCTCCGTGGGCGACAGAGTGACCATCACATGCAAGGCCTCCCAGAACGTGG
ACACCAACGTGGCATGGTATCAGCAGAAGCCAGGCCAGGCCCCTAAGTCCCTGAT
CTACTCTGCCTCCTACCGGTACTCCGACGTGCCCTCCAGGTTCTCTGGCTCCGCCT
CTGGCACCGACTTCACCCTGACCATCTCTTCCGTGCAGTCCGAGGACTTCGCTACC
TACTACTGCCAGCAGTACGACTCCTACCCTTACACCTTCGGCGGAGGCACCAAGCT GGAAATCAAG
822. PM76B10 HL .times. I2C artificial nt
CAGGTGCAGCTGGTCGAGTCTGGCGGCGGACTGGTCAAGCCTGGCGAGTCCCTGA HL codon
optimized GACTGTCTTGCGCTGCCTCCGGCTTCACCTTCTCCGACTACTACATGTACTGGGTC
CGCCAGGCTCCTGGCAAGGGACTGGAATGGGTGGCCATCATCTCCGACGGCGGCT
ACTACACCTACTACTCCGACATCATCAAGGGCCGGTTCACCATCTCCAGGGACAAC
GCCAAGAACTCCCTGTACCTGCAGATGAACTCCCTGAAGGCCGAGGACACCGCCG
TGTACTACTGCGCCAGGGGCTTCCCACTGCTGAGACACGGCGCCATGGATTACTG
GGGCCAGGGCACCCTGGTCACAGTGTCCTCTGGCGGAGGCGGAAGTGGAGGCGG
AGGAAGCGGAGGCGGCGGATCCGACATCCAGATGACCCAGTCCCCATCCTCCCTG
TCTGCCTCCGTGGGCGACAGAGTGACCATCACATGCAAGGCCTCCCAGAACGTGG
ACACCAACGTGGCATGGTATCAGCAGAAGCCAGGCCAGGCCCCTAAGTCCCTGAT
CTACTCTGCCTCCTACCGGTACTCCGACGTGCCCTCCAGGTTCTCTGGCTCCGCCT
CTGGCACCGACTTCACCCTGACCATCTCTTCCGTGCAGTCCGAGGACTTCGCTACC
TACTACTGCCAGCAGTACGACTCCTACCCTTACACCTTCGGCGGAGGCACCAAGCT
GGAAATCAAGTCCGGCGGAGGGGGCTCTGAAGTGCAGCTGGTGGAAAGCGGAGG
GGGACTGGTGCAGCCCGGGGGAAGTCTGAAGCTGTCCTGTGCCGCCAGCGGCTTT
ACCTTCAACAAGTACGCCATGAATTGGGTCCGACAGGCCCCAGGGAAAGGCCTGG
AATGGGTGGCACGGATCCGGTCCAAGTACAACAACTACGCCACCTACTACGCTGAC
TCCGTGAAGGACAGATTCACCATCAGCCGGGACGACTCTAAGAACACCGCCTATCT
GCAGATGAACAACCTGAAAACCGAGGATACAGCTGTGTACTATTGTGTGCGGCACG
GCAACTTCGGCAACTCCTACATCTCCTACTGGGCCTATTGGGGACAGGGAACACTG
GTCACCGTGTCTAGCGGAGGTGGCGGAAGTGGGGGAGGCGGATCTGGCGGTGGC
GGATCCCAGACCGTGGTCACCCAGGAACCTTCCCTGACCGTCTCCCCAGGCGGCA
CCGTGACCCTGACCTGTGGCTCCTCTACCGGCGCTGTGACCTCCGGCAACTACCC
TAACTGGGTGCAGCAGAAACCCGGACAGGCTCCTAGAGGCCTGATCGGCGGCACC
AAGTTTCTGGCCCCTGGCACCCCTGCCAGATTCTCCGGCTCCCTGCTGGGAGGCA
AGGCCGCTCTGACCCTGTCTGGCGTGCAGCCTGAGGACGAGGCCGAGTACTACTG
TGTGCTGTGGTACTCCAACAGATGGGTGTTCGGAGGCGGCACAAAGCTGACCGTG CTG 823.
PM34C7-H artificial aa
EVQLLEQSGAELVKPGASVKLSCTASGFNIKDTYMDWVKQRPEQGLEWIARIDPANGD
SKYDPKFQGKATITADTSSNTAYLQLSSLTSEDTAVYYCARGGMIWYFDVWGQGTTVT VSS 824.
PM34C7-HCDR1 artificial aa DTYMD 825. PM34C7-HCDR2 artificial aa
RIDPANGDSKYDPKFQG 826. PM34C7-HCDR3 artificial aa GGMIWYFDV 827.
PM34C7-H artificial nt
GAGGTGCAGCTGCTCGAGCAGTCTGGGGCAGAGCTTGTGAAGCCAGGGGCCTCA
GTCAAGTTGTCCTGCACAGCTTCTGGCTTCAACATTAAAGACACCTATATGGACTGG
GTGAAGCAGAGGCCTGAACAGGGCCTGGAATGGATTGCAAGGATTGATCCTGCGA
ATGGTGATAGTAAATATGACCCGAAATTCCAGGGCAAGGCCACTATAACAGCAGAC
ACATCCTCCAACACAGCCTACCTGCAGCTCAGCAGCCTGACATCTGAGGACACTGC
CGTCTATTATTGTGCTAGAGGCGGGATGATATGGTACTTCGATGTCTGGGGCCAAG
GGACCACGGTCACCGTCTCCTCA 828. PM34C7-L artificial aa
ELVLTQSPTTMAASPGEKITITCSASSSISSNYLHWYQQKPGFSPKLLIYRTSNLASGVP
ARFSGSGSGTSYSLTIGTMEAEDVATYYCQQGSSLPYTFGGGTKLEIK 829. PM34C7-LCDR1
artificial aa SASSSISSNYLH 830. PM34C7-LCDR2 artificial aa RTSNLAS
831. PM34C7-LCDR3 artificial aa QQGSSLPYT 832. PM34C7-L artificial
nt GAGCTCGTGCTCACCCAGTCTCCAACCACCATGGCTGCATCTCCCGGGGAGAAGA
TCACTATCACCTGCAGTGCCAGCTCAAGTATAAGTTCCAATTACTTGCATTGGTATC
AGCAGAAGCCAGGATTCTCCCCTAAACTCTTGATTTATAGGACATCCAATCTGGCTT
CTGGAGTCCCAGCTCGCTTCAGTGGCAGTGGGTCTGGGACCTCTTACTCTCTCACA
ATTGGCACCATGGAGGCTGAAGATGTTGCCACTTACTACTGCCAGCAGGGTAGTAG
TTTACCGTACACGTTCGGAGGGGGGACCAAGCTTGAGATCAAA 833. PM34C7-HL
artificial aa
EVQLLEQSGAELVKPGASVKLSCTASGFNIKDTYMDWVKQRPEQGLEWIARIDPANGD
SKYDPKFQGKATITADTSSNTAYLQLSSLTSEDTAVYYCARGGMIWYFDVWGQGTTVT
VSSGGGGSGGGGSGGGGSELVLTQSPTTMAASPGEKITITCSASSSISSNYLHWYQQK
PGFSPKLLIYRTSNLASGVPARFSGSGSGTSYSLTIGTMEAEDVATYYCQQGSSLPYTF
GGGTKLEIK 834. PM34C7-HL artificial nt
GAGGTGCAGCTGCTCGAGCAGTCTGGGGCAGAGCTTGTGAAGCCAGGGGCCTCA
GTCAAGTTGTCCTGCACAGCTTCTGGCTTCAACATTAAAGACACCTATATGGACTGG
GTGAAGCAGAGGCCTGAACAGGGCCTGGAATGGATTGCAAGGATTGATCCTGCGA
ATGGTGATAGTAAATATGACCCGAAATTCCAGGGCAAGGCCACTATAACAGCAGAC
ACATCCTCCAACACAGCCTACCTGCAGCTCAGCAGCCTGACATCTGAGGACACTGC
CGTCTATTATTGTGCTAGAGGCGGGATGATATGGTACTTCGATGTCTGGGGCCAAG
GGACCACGGTCACCGTCTCCTCAGGTGGTGGTGGTTCTGGCGGCGGCGGCTCCG
GTGGTGGTGGTTCTGAGCTCGTGCTCACCCAGTCTCCAACCACCATGGCTGCATCT
CCCGGGGAGAAGATCACTATCACCTGCAGTGCCAGCTCAAGTATAAGTTCCAATTA
CTTGCATTGGTATCAGCAGAAGCCAGGATTCTCCCCTAAACTCTTGATTTATAGGAC
ATCCAATCTGGCTTCTGGAGTCCCAGCTCGCTTCAGTGGCAGTGGGTCTGGGACCT
CTTACTCTCTCACAATTGGCACCATGGAGGCTGAAGATGTTGCCACTTACTACTGCC
AGCAGGGTAGTAGTTTACCGTACACGTTCGGAGGGGGGACCAAGCTTGAGATCAAA 835.
PM34C7 HL .times. I2C artificial aa
EVQLLEQSGAELVKPGASVKLSCTASGFNIKDTYMDWVKQRPEQGLEWIARIDPANGD HL
SKYDPKFQGKATITADTSSNTAYLQLSSLTSEDTAVYYCARGGMIWYFDVWGQGTTVT
VSSGGGGSGGGGSGGGGSELVLTQSPTTMAASPGEKITITCSASSSISSNYLHWYQQK
PGFSPKLLIYRTSNLASGVPARFSGSGSGTSYSLTIGTMEAEDVATYYCQQGSSLPYTF
GGGTKLEIKSGGGGSEVQLVESGGGLVQPGGSLKLSCAASGFTFNKYAMNWVRQAPG
KGLEWVARIRSKYNNYATYYADSVKDRFTISRDDSKNTAYLQMNNLKTEDTAVYYCVR
HGNFGNSYISYWAYWGQGTLVTVSSGGGGSGGGGSGGGGSQTVVTQEPSLTVSPG
GTVTLTCGSSTGAVTSGNYPNWVQQKPGQAPRGLIGGTKFLAPGTPARFSGSLLGGK
AALTLSGVQPEDEAEYYCVLWYSNRWVFGGGTKLTVL 836. PM34C7 HL .times. I2C
artificial nt
GAGGTGCAGCTGCTCGAGCAGTCTGGGGCAGAGCTTGTGAAGCCAGGGGCCTCA HL
GTCAAGTTGTCCTGCACAGCTTCTGGCTTCAACATTAAAGACACCTATATGGACTGG
GTGAAGCAGAGGCCTGAACAGGGCCTGGAATGGATTGCAAGGATTGATCCTGCGA
ATGGTGATAGTAAATATGACCCGAAATTCCAGGGCAAGGCCACTATAACAGCAGAC
ACATCCTCCAACACAGCCTACCTGCAGCTCAGCAGCCTGACATCTGAGGACACTGC
CGTCTATTATTGTGCTAGAGGCGGGATGATATGGTACTTCGATGTCTGGGGCCAAG
GGACCACGGTCACCGTCTCCTCAGGTGGTGGTGGTTCTGGCGGCGGCGGCTCCG
GTGGTGGTGGTTCTGAGCTCGTGCTCACCCAGTCTCCAACCACCATGGCTGCATCT
CCCGGGGAGAAGATCACTATCACCTGCAGTGCCAGCTCAAGTATAAGTTCCAATTA
CTTGCATTGGTATCAGCAGAAGCCAGGATTCTCCCCTAAACTCTTGATTTATAGGAC
ATCCAATCTGGCTTCTGGAGTCCCAGCTCGCTTCAGTGGCAGTGGGTCTGGGACCT
CTTACTCTCTCACAATTGGCACCATGGAGGCTGAAGATGTTGCCACTTACTACTGCC
AGCAGGGTAGTAGTTTACCGTACACGTTCGGAGGGGGGACCAAGCTTGAGATCAAA
TCCGGAGGTGGTGGATCCGAGGTGCAGCTGGTCGAGTCTGGAGGAGGATTGGTGC
AGCCTGGAGGGTCATTGAAACTCTCATGTGCAGCCTCTGGATTCACCTTCAATAAGT
ACGCCATGAACTGGGTCCGCCAGGCTCCAGGAAAGGGTTTGGAATGGGTTGCTCG
CATAAGAAGTAAATATAATAATTATGCAACATATTATGCCGATTCAGTGAAAGACAGG
TTCACCATCTCCAGAGATGATTCAAAAAACACTGCCTATCTACAAATGAACAACTTGA
AAACTGAGGACACTGCCGTGTACTACTGTGTGAGACATGGGAACTTCGGTAATAGC
TACATATCCTACTGGGCTTACTGGGGCCAAGGGACTCTGGTCACCGTCTCCTCAGG
TGGTGGTGGTTCTGGCGGCGGCGGCTCCGGTGGTGGTGGTTCTCAGACTGTTGTG
ACTCAGGAACCTTCACTCACCGTATCACCTGGTGGAACAGTCACACTCACTTGTGG
CTCCTCGACTGGGGCTGTTACATCTGGCAACTACCCAAACTGGGTCCAACAAAAAC
CAGGTCAGGCACCCCGTGGTCTAATAGGTGGGACTAAGTTCCTCGCCCCCGGTAC
TCCTGCCAGATTCTCAGGCTCCCTGCTTGGAGGCAAGGCTGCCCTCACCCTCTCAG
GGGTACAGCCAGAGGATGAGGCAGAATATTACTGTGTTCTATGGTACAGCAACCGC
TGGGTGTTCGGTGGAGGAACCAAACTGACTGTCCTA 837. PM49B9-H artificial aa
QVQLVQSGAEVKKPGASVKLSCKASGYTFTYFDINWVRQTPEQGLEWMGGISPGDGN
TNYNENFKGRVTMTRDTSNSTAYMELSRLRSDDTAVYYCARDGNFPYYAMDSWGQG
TTVTVSS 838. PM49B9-HCDR1 artificial aa YFDIN 839. PM49B9-HCDR2
artificial aa GISPGDGNTNYNENFKG 840. PM49B9-HCDR3 artificial aa
DGNFPYYAMDS 841. PM49B9-H artificial nt
CAGGTGCAGCTGGTCCAGTCTGGCGCCGAAGTGAAGAAGCCTGGCGCCTCCGTGA
AGCTGTCCTGCAAGGCCTCCGGCTACACCTTCACCTACTTCGACATCAACTGGGTG
CGGCAGACGCCTGAGCAGGGCCTGGAATGGATGGGCGGCATCTCCCCTGGCGAC
GGCAACACCAACTACAACGAGAACTTCAAGGGCAGGGTCACAATGACCAGAGACAC
GTCCAACTCCACCGCCTACATGGAGCTGTCCCGGCTGAGATCTGACGACACCGCC
GTGTACTACTGCGCCAGGGACGGCAACTTCCCTTACTACGCCATGGACTCTTGGGG
CCAGGGCACCACGGTCACCGTCTCCTCA 842. PM49B9-L artificial aa
DIVMTQTPLSLPVTLGDPASISCRSSQSLVYSNGNTYLNWYQQRPGQSPRLLIYKVSNR
FSGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCSQSTHVPYTFGQGTKLEIK 843.
PM49B9-LCDR1 artificial aa RSSQSLVYSNGNTYLN 844. PM49B9-LCDR2
artificial aa KVSNRFS 845. PM49B9-LCDR3 artificial aa SQSTHVPYT
846. PM49B9-L artificial nt
GACGTCGTGATGACTCAGACTCCACTCTCCCTGCCCGTCACCCTTGGAGACCCGG
CCTCCATCTCCTGCAGGTCTAGTCAAAGCCTCGTATACAGTAACGGAAACACCTACT
TGAATTGGTATCAACAGAGGCCAGGCCAATCTCCAAGACTCCTAATTTATAAGGTTT
CTAACCGGTTCTCTGGGGTCCCAGACAGATTCAGCGGCAGTGGGTCAGGCACTGA
TTTCACACTGAAAATCAGCAGGGTGGAGGCTGAGGATGTTGGGGTTTACTACTGCT
CTCAAAGTACACATGTTCCGTACACGTTTGGCCAGGGGACCAAGCTGGAGATCAAA 847.
PM49B9-HL artificial aa
QVQLVQSGAEVKKPGASVKLSCKASGYTFTYFDINWVRQTPEQGLEWMGGISPGDGN
TNYNENFKGRVTMTRDTSNSTAYMELSRLRSDDTAVYYCARDGNFPYYAMDSWGQG
TTVTVSSGGGGSGGGGSGGGGSDIVMTQTPLSLPVTLGDPASISCRSSQSLVYSNGNT
YLNWYQQRPGQSPRLLIYKVSNRFSGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCS
QSTHVPYTFGQGTKLEIK 848. PM49B9-HL artificial nt
CAGGTGCAGCTGGTCCAGTCTGGCGCCGAAGTGAAGAAGCCTGGCGCCTCCGTGA
AGCTGTCCTGCAAGGCCTCCGGCTACACCTTCACCTACTTCGACATCAACTGGGTG
CGGCAGACGCCTGAGCAGGGCCTGGAATGGATGGGCGGCATCTCCCCTGGCGAC
GGCAACACCAACTACAACGAGAACTTCAAGGGCAGGGTCACAATGACCAGAGACAC
GTCCAACTCCACCGCCTACATGGAGCTGTCCCGGCTGAGATCTGACGACACCGCC
GTGTACTACTGCGCCAGGGACGGCAACTTCCCTTACTACGCCATGGACTCTTGGGG
CCAGGGCACCACGGTCACCGTCTCCTCAGGTGGTGGTGGTTCTGGCGGCGGCGG
CTCCGGTGGTGGTGGTTCTGACGTCGTGATGACTCAGACTCCACTCTCCCTGCCCG
TCACCCTTGGAGACCCGGCCTCCATCTCCTGCAGGTCTAGTCAAAGCCTCGTATAC
AGTAACGGAAACACCTACTTGAATTGGTATCAACAGAGGCCAGGCCAATCTCCAAG
ACTCCTAATTTATAAGGTTTCTAACCGGTTCTCTGGGGTCCCAGACAGATTCAGCGG
CAGTGGGTCAGGCACTGATTTCACACTGAAAATCAGCAGGGTGGAGGCTGAGGAT
GTTGGGGTTTACTACTGCTCTCAAAGTACACATGTTCCGTACACGTTTGGCCAGGG
GACCAAGCTGGAGATCAAA 849. PM49B9 HL .times. I2C artificial aa
QVQLVQSGAEVKKPGASVKLSCKASGYTFTYFDINWVRQTPEQGLEWMGGISPGDGN HL
TNYNENFKGRVTMTRDTSNSTAYMELSRLRSDDTAVYYCARDGNFPYYAMDSWGQG
TTVTVSSGGGGSGGGGSGGGGSDIVMTQTPLSLPVTLGDPASISCRSSQSLVYSNGNT
YLNWYQQRPGQSPRLLIYKVSNRFSGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCS
QSTHVPYTFGQGTKLEIKSGGGGSEVQLVESGGGLVQPGGSLKLSCAASGFTFNKYA
MNWVRQAPGKGLEWVARIRSKYNNYATYYADSVKDRFTISRDDSKNTAYLQMNNLKT
EDTAVYYCVRHGNFGNSYISYWAYWGQGTLVTVSSGGGGSGGGGSGGGGSQTVVT
QEPSLTVSPGGTVTLTCGSSTGAVTSGNYPNWVQQKPGQAPRGLIGGTKFLAPGTPA
RFSGSLLGGKAALTLSGVQPEDEAEYYCVLWYSNRWVFGGGTKLTVL 850. PM49B9 HL
.times. I2C artificial nt
CAGGTGCAGCTGGTCCAGTCTGGCGCCGAAGTGAAGAAGCCTGGCGCCTCCGTGA HL
AGCTGTCCTGCAAGGCCTCCGGCTACACCTTCACCTACTTCGACATCAACTGGGTG
CGGCAGACGCCTGAGCAGGGCCTGGAATGGATGGGCGGCATCTCCCCTGGCGAC
GGCAACACCAACTACAACGAGAACTTCAAGGGCAGGGTCACAATGACCAGAGACAC
GTCCAACTCCACCGCCTACATGGAGCTGTCCCGGCTGAGATCTGACGACACCGCC
GTGTACTACTGCGCCAGGGACGGCAACTTCCCTTACTACGCCATGGACTCTTGGGG
CCAGGGCACCACGGTCACCGTCTCCTCAGGTGGTGGTGGTTCTGGCGGCGGCGG
CTCCGGTGGTGGTGGTTCTGACGTCGTGATGACTCAGACTCCACTCTCCCTGCCCG
TCACCCTTGGAGACCCGGCCTCCATCTCCTGCAGGTCTAGTCAAAGCCTCGTATAC
AGTAACGGAAACACCTACTTGAATTGGTATCAACAGAGGCCAGGCCAATCTCCAAG
ACTCCTAATTTATAAGGTTTCTAACCGGTTCTCTGGGGTCCCAGACAGATTCAGCGG
CAGTGGGTCAGGCACTGATTTCACACTGAAAATCAGCAGGGTGGAGGCTGAGGAT
GTTGGGGTTTACTACTGCTCTCAAAGTACACATGTTCCGTACACGTTTGGCCAGGG
GACCAAGCTGGAGATCAAATCCGGAGGTGGTGGATCCGAGGTGCAGCTGGTCGAG
TCTGGAGGAGGATTGGTGCAGCCTGGAGGGTCATTGAAACTCTCATGTGCAGCCTC
TGGATTCACCTTCAATAAGTACGCCATGAACTGGGTCCGCCAGGCTCCAGGAAAGG
GTTTGGAATGGGTTGCTCGCATAAGAAGTAAATATAATAATTATGCAACATATTATGC
CGATTCAGTGAAAGACAGGTTCACCATCTCCAGAGATGATTCAAAAAACACTGCCTA
TCTACAAATGAACAACTTGAAAACTGAGGACACTGCCGTGTACTACTGTGTGAGACA
TGGGAACTTCGGTAATAGCTACATATCCTACTGGGCTTACTGGGGCCAAGGGACTC
TGGTCACCGTCTCCTCAGGTGGTGGTGGTTCTGGCGGCGGCGGCTCCGGTGGTG
GTGGTTCTCAGACTGTTGTGACTCAGGAACCTTCACTCACCGTATCACCTGGTGGA
ACAGTCACACTCACTTGTGGCTCCTCGACTGGGGCTGTTACATCTGGCAACTACCC
AAACTGGGTCCAACAAAAACCAGGTCAGGCACCCCGTGGTCTAATAGGTGGGACTA
AGTTCCTCGCCCCCGGTACTCCTGCCAGATTCTCAGGCTCCCTGCTTGGAGGCAAG
GCTGCCCTCACCCTCTCAGGGGTACAGCCAGAGGATGAGGCAGAATATTACTGTGT
TCTATGGTACAGCAACCGCTGGGTGTTCGGTGGAGGAACCAAACTGACTGTCCTA 851.
PM29G1-H artificial aa
QVQLVQSGAEVKKPGASVKLSCKASGYTFTYFDINWVRQTPEQGLEWMGGISPGDGN
TNYNENFKGRVTMTRDTSNSTAYMELSRLRSDDTAVYYCARDGNFPYYAMDSWGQG TTVTVSS
852. PM29G1-HCDR1 artificial aa YFDIN 853. PM29G1-HCDR2 artificial
aa GISPGDGNTNYNENFKG 854. PM29G1-HCDR3 artificial aa DGNFPYYAMDS
855. PM29G1-H artificial nt
CAGGTGCAGCTGGTCCAGTCTGGCGCCGAAGTGAAGAAGCCTGGCGCCTCCGTGA
AGCTGTCCTGCAAGGCCTCCGGCTACACCTTCACCTACTTCGACATCAACTGGGTG
CGGCAGACGCCTGAGCAGGGCCTGGAATGGATGGGCGGCATCTCCCCTGGCGAC
GGCAACACCAACTACAACGAGAACTTCAAGGGCAGGGTCACAATGACCAGAGACAC
GTCCAACTCCACCGCCTACATGGAGCTGTCCCGGCTGAGATCTGACGACACCGCC
GTGTACTACTGCGCCAGGGACGGCAACTTCCCTTACTACGCCATGGACTCTTGGGG
CCAGGGCACCACGGTCACCGTCTCCTCA 856. PM29G1-L artificial aa
DVVMTQSPLSLPVTLGEPASISCRSSQSLVYSNGNTYLHWYQQKPGQSPRLLIYKVSN
RFSGVPDRFSGSGSGTDFTLKISRVEAEDVGVYFCSQSTHVPYTFGQGTKLEIK 857.
PM29G1-LCDR1 artificial aa RSSQSLVYSNGNTYLH 858. PM29G1-LCDR2
artificial aa KVSNRFS 859. PM29G1-LCDR3 artificial aa SQSTHVPYT
860. PM29G1-L artificial nt
GACGTCGTGATGACTCAGTCTCCACTCTCCCTGCCCGTCACCCTTGGAGAGCCGG
CCTCCATCTCCTGCAGGTCTAGTCAAAGCCTCGTATACAGTAACGGAAACACCTACT
TGCATTGGTATCAACAGAAGCCAGGCCAATCTCCAAGACTCCTAATTTATAAGGTTT
CTAACCGGTTCTCTGGGGTCCCAGACAGATTCAGCGGCAGTGGGTCAGGCACTGA
TTTCACACTGAAAATCAGCAGGGTGGAGGCTGAGGATGTTGGGGTTTATTTCTGCT
CTCAAAGTACACATGTTCCGTACACGTTTGGCCAGGGGACCAAGCTGGAGATCAAA 861.
PM29G1-HL artificial aa
QVQLVQSGAEVKKPGASVKLSCKASGYTFTYFDINWVRQTPEQGLEWMGGISPGDGN
TNYNENFKGRVTMTRDTSNSTAYMELSRLRSDDTAVYYCARDGNFPYYAMDSWGQG
TTVTVSSGGGGSGGGGSGGGGSDVVMTQSPLSLPVTLGEPASISCRSSQSLVYSNGN
TYLHWYQQKPGQSPRLLIYKVSNRFSGVPDRFSGSGSGTDFTLKISRVEAEDVGVYFC
SQSTHVPYTFGQGTKLEIK 862. PM29G1-HL artificial nt
CAGGTGCAGCTGGTCCAGTCTGGCGCCGAAGTGAAGAAGCCTGGCGCCTCCGTGA
AGCTGTCCTGCAAGGCCTCCGGCTACACCTTCACCTACTTCGACATCAACTGGGTG
CGGCAGACGCCTGAGCAGGGCCTGGAATGGATGGGCGGCATCTCCCCTGGCGAC
GGCAACACCAACTACAACGAGAACTTCAAGGGCAGGGTCACAATGACCAGAGACAC
GTCCAACTCCACCGCCTACATGGAGCTGTCCCGGCTGAGATCTGACGACACCGCC
GTGTACTACTGCGCCAGGGACGGCAACTTCCCTTACTACGCCATGGACTCTTGGGG
CCAGGGCACCACGGTCACCGTCTCCTCAGGTGGTGGTGGTTCTGGCGGCGGCGG
CTCCGGTGGTGGTGGTTCTGACGTCGTGATGACTCAGTCTCCACTCTCCCTGCCCG
TCACCCTTGGAGAGCCGGCCTCCATCTCCTGCAGGTCTAGTCAAAGCCTCGTATAC
AGTAACGGAAACACCTACTTGCATTGGTATCAACAGAAGCCAGGCCAATCTCCAAG
ACTCCTAATTTATAAGGTTTCTAACCGGTTCTCTGGGGTCCCAGACAGATTCAGCGG
CAGTGGGTCAGGCACTGATTTCACACTGAAAATCAGCAGGGTGGAGGCTGAGGAT
GTTGGGGTTTATTTCTGCTCTCAAAGTACACATGTTCCGTACACGTTTGGCCAGGGG
ACCAAGCTGGAGATCAAA 863. PM29G1 HL .times. I2C artificial aa
QVQLVQSGAEVKKPGASVKLSCKASGYTFTYFDINWVRQTPEQGLEWMGGISPGDGN HL
TNYNENFKGRVTMTRDTSNSTAYMELSRLRSDDTAVYYCARDGNFPYYAMDSWGQG
TTVTVSSGGGGSGGGGSGGGGSDVVMTQSPLSLPVTLGEPASISCRSSQSLVYSNGN
TYLHWYQQKPGQSPRLLIYKVSNRFSGVPDRFSGSGSGTDFTLKISRVEAEDVGVYFC
SQSTHVPYTFGQGTKLEIKSGGGGSEVQLVESGGGLVQPGGSLKLSCAASGFTFNKYA
MNWVRQAPGKGLEWVARIRSKYNNYATYYADSVKDRFTISRDDSKNTAYLQMNNLKT
EDTAVYYCVRHGNFGNSYISYWAYWGQGTLVTVSSGGGGSGGGGSGGGGSQTVVT
QEPSLTVSPGGTVTLTCGSSTGAVTSGNYPNWVQQKPGQAPRGLIGGTKFLAPGTPA
RFSGSLLGGKAALTLSGVQPEDEAEYYCVLWYSNRWVFGGGTKLTVL 864. PM29G1 HL
.times. I2C artificial nt
CAGGTGCAGCTGGTCCAGTCTGGCGCCGAAGTGAAGAAGCCTGGCGCCTCCGTGA HL
AGCTGTCCTGCAAGGCCTCCGGCTACACCTTCACCTACTTCGACATCAACTGGGTG
CGGCAGACGCCTGAGCAGGGCCTGGAATGGATGGGCGGCATCTCCCCTGGCGAC
GGCAACACCAACTACAACGAGAACTTCAAGGGCAGGGTCACAATGACCAGAGACAC
GTCCAACTCCACCGCCTACATGGAGCTGTCCCGGCTGAGATCTGACGACACCGCC
GTGTACTACTGCGCCAGGGACGGCAACTTCCCTTACTACGCCATGGACTCTTGGGG
CCAGGGCACCACGGTCACCGTCTCCTCAGGTGGTGGTGGTTCTGGCGGCGGCGG
CTCCGGTGGTGGTGGTTCTGACGTCGTGATGACTCAGTCTCCACTCTCCCTGCCCG
TCACCCTTGGAGAGCCGGCCTCCATCTCCTGCAGGTCTAGTCAAAGCCTCGTATAC
AGTAACGGAAACACCTACTTGCATTGGTATCAACAGAAGCCAGGCCAATCTCCAAG
ACTCCTAATTTATAAGGTTTCTAACCGGTTCTCTGGGGTCCCAGACAGATTCAGCGG
CAGTGGGTCAGGCACTGATTTCACACTGAAAATCAGCAGGGTGGAGGCTGAGGAT
GTTGGGGTTTATTTCTGCTCTCAAAGTACACATGTTCCGTACACGTTTGGCCAGGGG
ACCAAGCTGGAGATCAAATCCGGAGGTGGTGGATCCGAGGTGCAGCTGGTCGAGT
CTGGAGGAGGATTGGTGCAGCCTGGAGGGTCATTGAAACTCTCATGTGCAGCCTCT
GGATTCACCTTCAATAAGTACGCCATGAACTGGGTCCGCCAGGCTCCAGGAAAGGG
TTTGGAATGGGTTGCTCGCATAAGAAGTAAATATAATAATTATGCAACATATTATGCC
GATTCAGTGAAAGACAGGTTCACCATCTCCAGAGATGATTCAAAAAACACTGCCTAT
CTACAAATGAACAACTTGAAAACTGAGGACACTGCCGTGTACTACTGTGTGAGACAT
GGGAACTTCGGTAATAGCTACATATCCTACTGGGCTTACTGGGGCCAAGGGACTCT
GGTCACCGTCTCCTCAGGTGGTGGTGGTTCTGGCGGCGGCGGCTCCGGTGGTGG
TGGTTCTCAGACTGTTGTGACTCAGGAACCTTCACTCACCGTATCACCTGGTGGAA
CAGTCACACTCACTTGTGGCTCCTCGACTGGGGCTGTTACATCTGGCAACTACCCA
AACTGGGTCCAACAAAAACCAGGTCAGGCACCCCGTGGTCTAATAGGTGGGACTAA
GTTCCTCGCCCCCGGTACTCCTGCCAGATTCTCAGGCTCCCTGCTTGGAGGCAAG
GCTGCCCTCACCCTCTCAGGGGTACAGCCAGAGGATGAGGCAGAATATTACTGTGT
TCTATGGTACAGCAACCGCTGGGTGTTCGGTGGAGGAACCAAACTGACTGTCCTA 865.
PM29G1-H artificial nt
CAGGTGCAGCTGGTCCAGTCAGGCGCCGAAGTGAAGAAACCTGGCGCCTCTGTGA codon
optimized AGCTGTCCTGCAAGGCCTCCGGCTACACCTTCACCTACTTCGACATCAACTGGGTC
CGCCAGACCCCTGAGCAGGGCCTGGAATGGATGGGCGGCATCTCCCCTGGCGAC
GGCAACACCAACTACAACGAGAACTTCAAGGGCAGAGTGACCATGACCAGGGACA
CCTCTAACTCCACCGCCTACATGGAACTGTCCCGGCTGAGATCCGACGACACCGCC
GTGTACTACTGCGCCAGGGACGGCAACTTCCCTTACTACGCCATGGACTCTTGGGG
CCAGGGCACCACCGTGACAGTGTCCTCT 866. PM29G1-L artificial nt
GATGTGGTCATGACCCAGTCCCCACTGTCCCTGCCTGTGACCCTGGGCGAGCCTG codon
optimized CCTCCATCTCCTGCCGGTCCTCCCAGTCCCTGGTGTACTCCAACGGCAATACCTAC
CTGCACTGGTATCAGCAGAAGCCTGGCCAGTCCCCTAGGCTGCTGATCTACAAGGT
GTCCAACCGGTTCTCCGGCGTGCCCGACAGATTCTCCGGCTCCGGCTCTGGCACC
GACTTCACACTGAAGATCTCCAGGGTGGAGGCTGAGGACGTGGGCGTGTACTTCT
GCTCCCAGTCCACCCACGTGCCCTACACCTTCGGACAGGGCACCAAGCTGGAAAT CAAG 867.
PM29G1-H artificial nt
CAGGTGCAGCTGGTCCAGTCAGGCGCCGAAGTGAAGAAACCTGGCGCCTCTGTGA codon
optimized AGCTGTCCTGCAAGGCCTCCGGCTACACCTTCACCTACTTCGACATCAACTGGGTC
CGCCAGACCCCTGAGCAGGGCCTGGAATGGATGGGCGGCATCTCCCCTGGCGAC
GGCAACACCAACTACAACGAGAACTTCAAGGGCAGAGTGACCATGACCAGGGACA
CCTCTAACTCCACCGCCTACATGGAACTGTCCCGGCTGAGATCCGACGACACCGCC
GTGTACTACTGCGCCAGGGACGGCAACTTCCCTTACTACGCCATGGACTCTTGGGG
CCAGGGCACCACCGTGACAGTGTCCTCTGGCGGCGGAGGAAGTGGAGGTGGAGG
ATCTGGCGGAGGCGGCTCCGATGTGGTCATGACCCAGTCCCCACTGTCCCTGCCT
GTGACCCTGGGCGAGCCTGCCTCCATCTCCTGCCGGTCCTCCCAGTCCCTGGTGT
ACTCCAACGGCAATACCTACCTGCACTGGTATCAGCAGAAGCCTGGCCAGTCCCCT
AGGCTGCTGATCTACAAGGTGTCCAACCGGTTCTCCGGCGTGCCCGACAGATTCTC
CGGCTCCGGCTCTGGCACCGACTTCACACTGAAGATCTCCAGGGTGGAGGCTGAG
GACGTGGGCGTGTACTTCTGCTCCCAGTCCACCCACGTGCCCTACACCTTCGGACA
GGGCACCAAGCTGGAAATCAAG 868. PM29G1 HL .times. I2C artificial nt
CAGGTGCAGCTGGTCCAGTCAGGCGCCGAAGTGAAGAAACCTGGCGCCTCTGTGA HL codon
optimized AGCTGTCCTGCAAGGCCTCCGGCTACACCTTCACCTACTTCGACATCAACTGGGTC
CGCCAGACCCCTGAGCAGGGCCTGGAATGGATGGGCGGCATCTCCCCTGGCGAC
GGCAACACCAACTACAACGAGAACTTCAAGGGCAGAGTGACCATGACCAGGGACA
CCTCTAACTCCACCGCCTACATGGAACTGTCCCGGCTGAGATCCGACGACACCGCC
GTGTACTACTGCGCCAGGGACGGCAACTTCCCTTACTACGCCATGGACTCTTGGGG
CCAGGGCACCACCGTGACAGTGTCCTCTGGCGGCGGAGGAAGTGGAGGTGGAGG
ATCTGGCGGAGGCGGCTCCGATGTGGTCATGACCCAGTCCCCACTGTCCCTGCCT
GTGACCCTGGGCGAGCCTGCCTCCATCTCCTGCCGGTCCTCCCAGTCCCTGGTGT
ACTCCAACGGCAATACCTACCTGCACTGGTATCAGCAGAAGCCTGGCCAGTCCCCT
AGGCTGCTGATCTACAAGGTGTCCAACCGGTTCTCCGGCGTGCCCGACAGATTCTC
CGGCTCCGGCTCTGGCACCGACTTCACACTGAAGATCTCCAGGGTGGAGGCTGAG
GACGTGGGCGTGTACTTCTGCTCCCAGTCCACCCACGTGCCCTACACCTTCGGACA
GGGCACCAAGCTGGAAATCAAGTCTGGCGGAGGGGGCTCTGAAGTGCAGCTGGTG
GAAAGCGGAGGGGGACTGGTGCAGCCCGGGGGAAGTCTGAAGCTGTCCTGTGCC
GCCAGCGGCTTTACCTTCAACAAGTACGCCATGAATTGGGTCCGACAGGCCCCAG
GGAAAGGCCTGGAATGGGTGGCACGGATCCGGTCCAAGTACAACAACTACGCCAC
CTACTACGCTGACTCCGTGAAGGACAGATTCACCATCAGCCGGGACGACTCTAAGA
ACACCGCCTATCTGCAGATGAACAACCTGAAAACCGAGGATACAGCTGTGTACTATT
GTGTGCGGCACGGCAACTTCGGCAACTCCTACATCTCCTACTGGGCCTATTGGGGA
CAGGGAACACTGGTCACCGTGTCTAGCGGAGGTGGCGGAAGTGGGGGAGGCGGA
TCTGGCGGTGGCGGATCCCAGACCGTGGTCACCCAGGAACCTTCCCTGACCGTCT
CCCCAGGCGGCACCGTGACCCTGACCTGTGGCTCCTCTACCGGCGCTGTGACCTC
CGGCAACTACCCTAACTGGGTGCAGCAGAAACCCGGACAGGCTCCTAGAGGCCTG
ATCGGCGGCACCAAGTTTCTGGCCCCTGGCACCCCTGCCAGATTCTCCGGCTCCC
TGCTGGGAGGCAAGGCCGCTCTGACCCTGTCTGGCGTGCAGCCTGAGGACGAGG
CCGAGTACTACTGTGTGCTGTGGTACTCCAACAGATGGGTGTTCGGAGGCGGCAC
AAAGCTGACCGTGCTG 869. PM08B6-H artificial aa
QVQLVQSGAEVMKPGASVKVSCKASGYTITDTYMDWVRQAPGQGLEWIARIDPANGD
SKYDPKFQGRVTMTADTSTNTVYMELSSLRSEDTAVYYCARGGMIWYFDVWGQGTTV TVSS 870.
PM08B6-HCDR1 artificial aa DTYMD 871. PM08B6-HCDR2 artificial aa
RIDPANGDSKYDPKFQG 872. PM08B6-HCDR3 artificial aa GGMIWYFDV 873.
PM08B6-H artificial nt
CAGGTGCAGCTGGTCCAGTCTGGGGCAGAGGTTATGAAACCAGGGGCCTCAGTCA
AGGTGTCCTGCAAAGCTTCTGGCTACACCATTACAGACACCTATATGGACTGGGTG
AGGCAGGCGCCTGGACAGGGCCTGGAATGGATTGCAAGGATTGATCCTGCGAATG
GTGATAGTAAATATGACCCGAAATTCCAGGGCAGGGTCACTATGACAGCAGACACA
TCCACCAACACAGTCTACATGGAGCTCAGCAGCCTGAGATCTGAGGACACTGCCGT
CTATTATTGTGCTAGAGGCGGGATGATATGGTACTTCGATGTCTGGGGCCAAGGGA
CCACGGTCACCGTCTCCTCA 874. PM08B6-L artificial aa
EIVLTQSPATLAVSPGEKVTLSCSASSSISSNYLHWYQQKPGLPPRLLIYRTSNLASGVP
DRFSGSGSGTDFTLTISRLEPEDFATYYCQQGSSLPYTFGQGTKLEIK 875. PM08B6-LCDR1
artificial aa SASSSISSNYLH 876. PM08B6-LCDR2 artificial aa RTSNLAS
877. PM08B6-LCDR3 artificial aa QQGSSLPYT 878. PM08B6-L artificial
nt GAGATCGTGCTCACCCAGTCTCCAGCCACCCTGGCTGTATCTCCCGGGGAGAAGG
TCACTCTCTCCTGCAGTGCCAGCTCAAGTATAAGTTCCAATTACTTGCATTGGTATC
AGCAGAAGCCAGGATTGCCCCCTAGACTCTTGATTTATAGGACATCCAATCTGGCTT
CTGGAGTCCCAGATCGCTTCAGTGGCAGTGGGTCTGGGACCGATTTCACTCTCACA
ATTAGCAGGCTGGAGCCTGAAGATTTTGCCACTTACTACTGCCAGCAGGGTAGTAG
TTTACCGTACACGTTCGGACAAGGGACCAAGCTTGAGATCAAA 879. PM08B6-HL
artificial aa
QVQLVQSGAEVMKPGASVKVSCKASGYTITDTYMDWVRQAPGQGLEWIARIDPANGD
SKYDPKFQGRVTMTADTSTNTVYMELSSLRSEDTAVYYCARGGMIWYFDVWGQGTTV
TVSSGGGGSGGGGSGGGGSEIVLTQSPATLAVSPGEKVTLSCSASSSISSNYLHWYQ
QKPGLPPRLLIYRTSNLASGVPDRFSGSGSGTDFTLTISRLEPEDFATYYCQQGSSLPY
TFGQGTKLEIK 880. PM08B6-HL artificial nt
CAGGTGCAGCTGGTCCAGTCTGGGGCAGAGGTTATGAAACCAGGGGCCTCAGTCA
AGGTGTCCTGCAAAGCTTCTGGCTACACCATTACAGACACCTATATGGACTGGGTG
AGGCAGGCGCCTGGACAGGGCCTGGAATGGATTGCAAGGATTGATCCTGCGAATG
GTGATAGTAAATATGACCCGAAATTCCAGGGCAGGGTCACTATGACAGCAGACACA
TCCACCAACACAGTCTACATGGAGCTCAGCAGCCTGAGATCTGAGGACACTGCCGT
CTATTATTGTGCTAGAGGCGGGATGATATGGTACTTCGATGTCTGGGGCCAAGGGA
CCACGGTCACCGTCTCCTCAGGTGGTGGTGGTTCTGGCGGCGGCGGCTCCGGTG
GTGGTGGTTCTGAGATCGTGCTCACCCAGTCTCCAGCCACCCTGGCTGTATCTCCC
GGGGAGAAGGTCACTCTCTCCTGCAGTGCCAGCTCAAGTATAAGTTCCAATTACTT
GCATTGGTATCAGCAGAAGCCAGGATTGCCCCCTAGACTCTTGATTTATAGGACATC
CAATCTGGCTTCTGGAGTCCCAGATCGCTTCAGTGGCAGTGGGTCTGGGACCGATT
TCACTCTCACAATTAGCAGGCTGGAGCCTGAAGATTTTGCCACTTACTACTGCCAGC
AGGGTAGTAGTTTACCGTACACGTTCGGACAAGGGACCAAGCTTGAGATCAAA 881. PM08B6
HL .times. I2C artificial aa
QVQLVQSGAEVMKPGASVKVSCKASGYTITDTYMDWVRQAPGQGLEWIARIDPANGD HL
SKYDPKFQGRVTMTADTSTNTVYMELSSLRSEDTAVYYCARGGMIWYFDVWGQGTTV
TVSSGGGGSGGGGSGGGGSEIVLTQSPATLAVSPGEKVTLSCSASSSISSNYLHWYQ
QKPGLPPRLLIYRTSNLASGVPDRFSGSGSGTDFTLTISRLEPEDFATYYCQQGSSLPY
TFGQGTKLEIKSGGGGSEVQLVESGGGLVQPGGSLKLSCAASGFTFNKYAMNWVRQA
PGKGLEWVARIRSKYNNYATYYADSVKDRFTISRDDSKNTAYLQMNNLKTEDTAVYYC
VRHGNFGNSYISYWAYWGQGTLVTVSSGGGGSGGGGSGGGGSQTVVTQEPSLTVSP
GGTVTLTCGSSTGAVTSGNYPNWVQQKPGQAPRGLIGGTKFLAPGTPARFSGSLLGG
KAALTLSGVQPEDEAEYYCVLWYSNRWVFGGGTKLTVL 882. PM08B6 HL .times. I2C
artificial nt
CAGGTGCAGCTGGTCCAGTCTGGGGCAGAGGTTATGAAACCAGGGGCCTCAGTCA HL
AGGTGTCCTGCAAAGCTTCTGGCTACACCATTACAGACACCTATATGGACTGGGTG
AGGCAGGCGCCTGGACAGGGCCTGGAATGGATTGCAAGGATTGATCCTGCGAATG
GTGATAGTAAATATGACCCGAAATTCCAGGGCAGGGTCACTATGACAGCAGACACA
TCCACCAACACAGTCTACATGGAGCTCAGCAGCCTGAGATCTGAGGACACTGCCGT
CTATTATTGTGCTAGAGGCGGGATGATATGGTACTTCGATGTCTGGGGCCAAGGGA
CCACGGTCACCGTCTCCTCAGGTGGTGGTGGTTCTGGCGGCGGCGGCTCCGGTG
GTGGTGGTTCTGAGATCGTGCTCACCCAGTCTCCAGCCACCCTGGCTGTATCTCCC
GGGGAGAAGGTCACTCTCTCCTGCAGTGCCAGCTCAAGTATAAGTTCCAATTACTT
GCATTGGTATCAGCAGAAGCCAGGATTGCCCCCTAGACTCTTGATTTATAGGACATC
CAATCTGGCTTCTGGAGTCCCAGATCGCTTCAGTGGCAGTGGGTCTGGGACCGATT
TCACTCTCACAATTAGCAGGCTGGAGCCTGAAGATTTTGCCACTTACTACTGCCAGC
AGGGTAGTAGTTTACCGTACACGTTCGGACAAGGGACCAAGCTTGAGATCAAATCC
GGAGGTGGTGGATCCGAGGTGCAGCTGGTCGAGTCTGGAGGAGGATTGGTGCAG
CCTGGAGGGTCATTGAAACTCTCATGTGCAGCCTCTGGATTCACCTTCAATAAGTAC
GCCATGAACTGGGTCCGCCAGGCTCCAGGAAAGGGTTTGGAATGGGTTGCTCGCA
TAAGAAGTAAATATAATAATTATGCAACATATTATGCCGATTCAGTGAAAGACAGGTT
CACCATCTCCAGAGATGATTCAAAAAACACTGCCTATCTACAAATGAACAACTTGAA
AACTGAGGACACTGCCGTGTACTACTGTGTGAGACATGGGAACTTCGGTAATAGCT
ACATATCCTACTGGGCTTACTGGGGCCAAGGGACTCTGGTCACCGTCTCCTCAGGT
GGTGGTGGTTCTGGCGGCGGCGGCTCCGGTGGTGGTGGTTCTCAGACTGTTGTGA
CTCAGGAACCTTCACTCACCGTATCACCTGGTGGAACAGTCACACTCACTTGTGGC
TCCTCGACTGGGGCTGTTACATCTGGCAACTACCCAAACTGGGTCCAACAAAAACC
AGGTCAGGCACCCCGTGGTCTAATAGGTGGGACTAAGTTCCTCGCCCCCGGTACT
CCTGCCAGATTCTCAGGCTCCCTGCTTGGAGGCAAGGCTGCCCTCACCCTCTCAG
GGGTACAGCCAGAGGATGAGGCAGAATATTACTGTGTTCTATGGTACAGCAACCGC
TGGGTGTTCGGTGGAGGAACCAAACTGACTGTCCTA 883. PM08B6-H artificial nt
CAGGTGCAGCTGGTCCAGTCTGGCGCCGAAGTGATGAAGCCTGGCGCCTCCGTGA codon
optimized AGGTGTCCTGCAAGGCCTCCGGCTACACCATCACCGACACCTACATGGACTGGGT
GCGGCAGGCTCCTGGACAGGGCCTGGAATGGATCGCCCGGATCGACCCTGCCAA
CGGCGACTCCAAGTACGACCCTAAGTTCCAGGGCAGAGTGACCATGACCGCCGAC
ACCTCCACCAACACCGTGTACATGGAACTGTCCTCCCTGCGGTCTGAGGACACCGC
CGTGTACTACTGCGCCAGGGGCGGCATGATCTGGTACTTCGACGTGTGGGGCCAG
GGCACCACCGTGACAGTGTCCTCT 884. PM08B6-L artificial nt
GAGATCGTGCTGACCCAGTCTCCTGCCACCCTGGCTGTGTCTCCCGGCGAGAAAG codon
optimized TGACCCTGTCCTGCTCCGCCTCCTCCTCCATCTCCTCCAACTACCTGCACTGGTATC
AGCAGAAGCCTGGCCTGCCTCCTCGGCTGCTGATCTACCGGACCTCCAACCTGGC
CTCTGGCGTGCCCGACAGGTTCTCCGGCTCTGGCTCCGGCACCGACTTCACCCTG
ACCATCTCCCGGCTGGAACCTGAGGACTTCGCCACCTACTACTGCCAGCAGGGCT
CCTCCCTGCCTTACACCTTCGGACAGGGCACCAAGCTGGAAATCAAG 885. PM08B6-HL
artificial nt
CAGGTGCAGCTGGTCCAGTCTGGCGCCGAAGTGATGAAGCCTGGCGCCTCCGTGA codon
optimized AGGTGTCCTGCAAGGCCTCCGGCTACACCATCACCGACACCTACATGGACTGGGT
GCGGCAGGCTCCTGGACAGGGCCTGGAATGGATCGCCCGGATCGACCCTGCCAA
CGGCGACTCCAAGTACGACCCTAAGTTCCAGGGCAGAGTGACCATGACCGCCGAC
ACCTCCACCAACACCGTGTACATGGAACTGTCCTCCCTGCGGTCTGAGGACACCGC
CGTGTACTACTGCGCCAGGGGCGGCATGATCTGGTACTTCGACGTGTGGGGCCAG
GGCACCACCGTGACAGTGTCCTCTGGCGGCGGAGGAAGTGGAGGTGGAGGATCT
GGTGGAGGCGGCTCCGAGATCGTGCTGACCCAGTCTCCTGCCACCCTGGCTGTGT
CTCCCGGCGAGAAAGTGACCCTGTCCTGCTCCGCCTCCTCCTCCATCTCCTCCAAC
TACCTGCACTGGTATCAGCAGAAGCCTGGCCTGCCTCCTCGGCTGCTGATCTACCG
GACCTCCAACCTGGCCTCTGGCGTGCCCGACAGGTTCTCCGGCTCTGGCTCCGGC
ACCGACTTCACCCTGACCATCTCCCGGCTGGAACCTGAGGACTTCGCCACCTACTA
CTGCCAGCAGGGCTCCTCCCTGCCTTACACCTTCGGACAGGGCACCAAGCTGGAA ATCAAG 886.
PM08B6 HL .times. I2C artificial nt
CAGGTGCAGCTGGTCCAGTCTGGCGCCGAAGTGATGAAGCCTGGCGCCTCCGTGA HL codon
optimized AGGTGTCCTGCAAGGCCTCCGGCTACACCATCACCGACACCTACATGGACTGGGT
GCGGCAGGCTCCTGGACAGGGCCTGGAATGGATCGCCCGGATCGACCCTGCCAA
CGGCGACTCCAAGTACGACCCTAAGTTCCAGGGCAGAGTGACCATGACCGCCGAC
ACCTCCACCAACACCGTGTACATGGAACTGTCCTCCCTGCGGTCTGAGGACACCGC
CGTGTACTACTGCGCCAGGGGCGGCATGATCTGGTACTTCGACGTGTGGGGCCAG
GGCACCACCGTGACAGTGTCCTCTGGCGGCGGAGGAAGTGGAGGTGGAGGATCT
GGTGGAGGCGGCTCCGAGATCGTGCTGACCCAGTCTCCTGCCACCCTGGCTGTGT
CTCCCGGCGAGAAAGTGACCCTGTCCTGCTCCGCCTCCTCCTCCATCTCCTCCAAC
TACCTGCACTGGTATCAGCAGAAGCCTGGCCTGCCTCCTCGGCTGCTGATCTACCG
GACCTCCAACCTGGCCTCTGGCGTGCCCGACAGGTTCTCCGGCTCTGGCTCCGGC
ACCGACTTCACCCTGACCATCTCCCGGCTGGAACCTGAGGACTTCGCCACCTACTA
CTGCCAGCAGGGCTCCTCCCTGCCTTACACCTTCGGACAGGGCACCAAGCTGGAA
ATCAAGTCTGGCGGAGGGGGCTCTGAAGTGCAGCTGGTGGAAAGCGGAGGGGGA
CTGGTGCAGCCCGGGGGAAGTCTGAAGCTGTCCTGTGCCGCCAGCGGCTTTACCT
TCAACAAGTACGCCATGAATTGGGTCCGACAGGCCCCAGGGAAAGGCCTGGAATG
GGTGGCACGGATCCGGTCCAAGTACAACAACTACGCCACCTACTACGCTGACTCCG
TGAAGGACAGATTCACCATCAGCCGGGACGACTCTAAGAACACCGCCTATCTGCAG
ATGAACAACCTGAAAACCGAGGATACAGCTGTGTACTATTGTGTGCGGCACGGCAA
CTTCGGCAACTCCTACATCTCCTACTGGGCCTATTGGGGACAGGGAACACTGGTCA
CCGTGTCTAGCGGAGGTGGCGGAAGTGGGGGAGGCGGATCTGGCGGTGGCGGAT
CCCAGACCGTGGTCACCCAGGAACCTTCCCTGACCGTCTCCCCAGGCGGCACCGT
GACCCTGACCTGTGGCTCCTCTACCGGCGCTGTGACCTCCGGCAACTACCCTAACT
GGGTGCAGCAGAAACCCGGACAGGCTCCTAGAGGCCTGATCGGCGGCACCAAGTT
TCTGGCCCCTGGCACCCCTGCCAGATTCTCCGGCTCCCTGCTGGGAGGCAAGGCC
GCTCTGACCCTGTCTGGCGTGCAGCCTGAGGACGAGGCCGAGTACTACTGTGTGC
TGTGGTACTCCAACAGATGGGTGTTCGGAGGCGGCACAAAGCTGACCGTGCTG 887.
PM08E11-H artificial aa
QVQLVQSGAEVMKPGASVKVSCKASGYTITDTYMDWVRQAPGQGLEWIARIDPANGD
SKYDPKFQGRVTMTADTSTNTVYMELSSLRSEDTAVYYCARGGMIWYFDVWGQGTTV TVSS 888.
PM08E11-HCDR1 artificial aa DTYMD 889. PM08E11-HCDR2 artificial aa
RIDPANGDSKYDPKFQG 890. PM08E11-HCDR3 artificial aa GGMIWYFDV 891.
PM08E11-H artificial nt
CAGGTGCAGCTGGTCCAGTCTGGGGCAGAGGTTATGAAACCAGGGGCCTCAGTCA
AGGTGTCCTGCAAAGCTTCTGGCTACACCATTACAGACACCTATATGGACTGGGTG
AGGCAGGCGCCTGGACAGGGCCTGGAATGGATTGCAAGGATTGATCCTGCGAATG
GTGATAGTAAATATGACCCGAAATTCCAGGGCAGGGTCACTATGACAGCAGACACA
TCCACCAACACAGTCTACATGGAGCTCAGCAGCCTGAGATCTGAGGACACTGCCGT
CTATTATTGTGCTAGAGGCGGGATGATATGGTACTTCGATGTCTGGGGCCAAGGGA
CCACGGTCACCGTCTCCTCA 892. PM08E11-L artificial aa
EIVLTQSPATMSVSPGERATLSCSASSSISSNYLHWYQQKPGLPPRLLIYRTSNLASGIP
DRFSGSGSGTDFTLTISRLEAEDFATYYCQQGSSLPYTFGQGTKLEIK 893. PM08E11-LCDR1
artificial aa SASSSISSNYLH 894. PM08E11-LCDR2 artificial aa RTSNLAS
895. PM08E11-LCDR3 artificial aa QQGSSLPYT 896. PM08E11-L
artificial nt
GAGATCGTGCTCACCCAGTCTCCAGCCACCATGTCTGTATCTCCCGGGGAGAGGG
CCACTCTCTCCTGCAGTGCCAGCTCAAGTATAAGTTCCAATTACTTGCATTGGTATC
AGCAGAAGCCAGGATTGCCCCCTAGACTCTTGATTTATAGGACATCCAATCTGGCTT
CTGGAATCCCAGATCGCTTCAGTGGCAGTGGGTCTGGGACCGATTTCACTCTCACA
ATTAGCAGGCTGGAGGCTGAAGATTTTGCCACTTACTACTGCCAGCAGGGTAGTAG
TTTACCGTACACGTTCGGACAAGGGACCAAGCTTGAGATCAAA 897. PM08E11-HL
artificial aa
QVQLVQSGAEVMKPGASVKVSCKASGYTITDTYMDWVRQAPGQGLEWIARIDPANGD
SKYDPKFQGRVTMTADTSTNTVYMELSSLRSEDTAVYYCARGGMIWYFDVWGQGTTV
TVSSGGGGSGGGGSGGGGSEIVLTQSPATMSVSPGERATLSCSASSSISSNYLHWYQ
QKPGLPPRLLIYRTSNLASGIPDRFSGSGSGTDFTLTISRLEAEDFATYYCQQGSSLPYT
FGQGTKLEIK 898. PM08E11-HL artificial nt
CAGGTGCAGCTGGTCCAGTCTGGGGCAGAGGTTATGAAACCAGGGGCCTCAGTCA
AGGTGTCCTGCAAAGCTTCTGGCTACACCATTACAGACACCTATATGGACTGGGTG
AGGCAGGCGCCTGGACAGGGCCTGGAATGGATTGCAAGGATTGATCCTGCGAATG
GTGATAGTAAATATGACCCGAAATTCCAGGGCAGGGTCACTATGACAGCAGACACA
TCCACCAACACAGTCTACATGGAGCTCAGCAGCCTGAGATCTGAGGACACTGCCGT
CTATTATTGTGCTAGAGGCGGGATGATATGGTACTTCGATGTCTGGGGCCAAGGGA
CCACGGTCACCGTCTCCTCAGGTGGTGGTGGTTCTGGCGGCGGCGGCTCCGGTG
GTGGTGGTTCTGAGATCGTGCTCACCCAGTCTCCAGCCACCATGTCTGTATCTCCC
GGGGAGAGGGCCACTCTCTCCTGCAGTGCCAGCTCAAGTATAAGTTCCAATTACTT
GCATTGGTATCAGCAGAAGCCAGGATTGCCCCCTAGACTCTTGATTTATAGGACATC
CAATCTGGCTTCTGGAATCCCAGATCGCTTCAGTGGCAGTGGGTCTGGGACCGATT
TCACTCTCACAATTAGCAGGCTGGAGGCTGAAGATTTTGCCACTTACTACTGCCAGC
AGGGTAGTAGTTTACCGTACACGTTCGGACAAGGGACCAAGCTTGAGATCAAA 899. PM08E11
HL .times. I2C artificial aa
QVQLVQSGAEVMKPGASVKVSCKASGYTITDTYMDWVRQAPGQGLEWIARIDPANGD HL
SKYDPKFQGRVTMTADTSTNTVYMELSSLRSEDTAVYYCARGGMIWYFDVWGQGTTV
TVSSGGGGSGGGGSGGGGSEIVLTQSPATMSVSPGERATLSCSASSSISSNYLHWYQ
QKPGLPPRLLIYRTSNLASGIPDRFSGSGSGTDFTLTISRLEAEDFATYYCQQGSSLPYT
FGQGTKLEIKSGGGGSEVQLVESGGGLVQPGGSLKLSCAASGFTFNKYAMNWVRQAP
GKGLEWVARIRSKYNNYATYYADSVKDRFTISRDDSKNTAYLQMNNLKTEDTAVYYCV
RHGNFGNSYISYWAYWGQGTLVTVSSGGGGSGGGGSGGGGSQTVVTQEPSLTVSPG
GTVTLTCGSSTGAVTSGNYPNWVQQKPGQAPRGLIGGTKFLAPGTPARFSGSLLGGK
AALTLSGVQPEDEAEYYCVLWYSNRWVFGGGTKLTVL 900. PM08E11 HL .times. I2C
artificial nt
CAGGTGCAGCTGGTCCAGTCTGGGGCAGAGGTTATGAAACCAGGGGCCTCAGTCA HL
AGGTGTCCTGCAAAGCTTCTGGCTACACCATTACAGACACCTATATGGACTGGGTG
AGGCAGGCGCCTGGACAGGGCCTGGAATGGATTGCAAGGATTGATCCTGCGAATG
GTGATAGTAAATATGACCCGAAATTCCAGGGCAGGGTCACTATGACAGCAGACACA
TCCACCAACACAGTCTACATGGAGCTCAGCAGCCTGAGATCTGAGGACACTGCCGT
CTATTATTGTGCTAGAGGCGGGATGATATGGTACTTCGATGTCTGGGGCCAAGGGA
CCACGGTCACCGTCTCCTCAGGTGGTGGTGGTTCTGGCGGCGGCGGCTCCGGTG
GTGGTGGTTCTGAGATCGTGCTCACCCAGTCTCCAGCCACCATGTCTGTATCTCCC
GGGGAGAGGGCCACTCTCTCCTGCAGTGCCAGCTCAAGTATAAGTTCCAATTACTT
GCATTGGTATCAGCAGAAGCCAGGATTGCCCCCTAGACTCTTGATTTATAGGACATC
CAATCTGGCTTCTGGAATCCCAGATCGCTTCAGTGGCAGTGGGTCTGGGACCGATT
TCACTCTCACAATTAGCAGGCTGGAGGCTGAAGATTTTGCCACTTACTACTGCCAGC
AGGGTAGTAGTTTACCGTACACGTTCGGACAAGGGACCAAGCTTGAGATCAAATCC
GGAGGTGGTGGATCCGAGGTGCAGCTGGTCGAGTCTGGAGGAGGATTGGTGCAG
CCTGGAGGGTCATTGAAACTCTCATGTGCAGCCTCTGGATTCACCTTCAATAAGTAC
GCCATGAACTGGGTCCGCCAGGCTCCAGGAAAGGGTTTGGAATGGGTTGCTCGCA
TAAGAAGTAAATATAATAATTATGCAACATATTATGCCGATTCAGTGAAAGACAGGTT
CACCATCTCCAGAGATGATTCAAAAAACACTGCCTATCTACAAATGAACAACTTGAA
AACTGAGGACACTGCCGTGTACTACTGTGTGAGACATGGGAACTTCGGTAATAGCT
ACATATCCTACTGGGCTTACTGGGGCCAAGGGACTCTGGTCACCGTCTCCTCAGGT
GGTGGTGGTTCTGGCGGCGGCGGCTCCGGTGGTGGTGGTTCTCAGACTGTTGTGA
CTCAGGAACCTTCACTCACCGTATCACCTGGTGGAACAGTCACACTCACTTGTGGC
TCCTCGACTGGGGCTGTTACATCTGGCAACTACCCAAACTGGGTCCAACAAAAACC
AGGTCAGGCACCCCGTGGTCTAATAGGTGGGACTAAGTTCCTCGCCCCCGGTACT
CCTGCCAGATTCTCAGGCTCCCTGCTTGGAGGCAAGGCTGCCCTCACCCTCTCAG
GGGTACAGCCAGAGGATGAGGCAGAATATTACTGTGTTCTATGGTACAGCAACCGC
TGGGTGTTCGGTGGAGGAACCAAACTGACTGTCCTA 901. PM08E11-H artificial nt
CAGGTGCAGCTGGTCCAGTCTGGCGCCGAAGTGATGAAGCCTGGCGCCTCCGTGA codon
optimized AGGTGTCCTGCAAGGCCTCCGGCTACACCATCACCGACACCTACATGGACTGGGT
GCGGCAGGCTCCTGGACAGGGCCTGGAATGGATCGCCCGGATCGACCCTGCCAA
CGGCGACTCCAAGTACGACCCTAAGTTCCAGGGCAGAGTGACCATGACCGCCGAC
ACCTCCACCAACACCGTGTACATGGAACTGTCCTCCCTGCGGTCTGAGGACACCGC
CGTGTACTACTGCGCCAGGGGCGGCATGATCTGGTACTTCGACGTGTGGGGCCAG
GGCACCACCGTGACAGTGTCCTCT 902. PM08E11-L artificial nt
GAGATCGTGCTGACCCAGTCTCCTGCCACCATGTCTGTGTCTCCCGGCGAGAGAG codon
optimized CCACCCTGTCCTGCTCCGCCTCCTCCTCCATCTCCTCCAACTACCTGCACTGGTAT
CAGCAGAAGCCTGGCCTGCCTCCTCGGCTGCTGATCTACCGGACCTCCAACCTGG
CCTCTGGCATCCCCGACAGGTTCTCCGGCTCTGGCTCCGGCACCGACTTCACCCT
GACCATCTCCCGGCTGGAAGCTGAGGACTTCGCCACCTACTACTGCCAGCAGGGC
TCCTCCCTGCCTTACACCTTCGGACAGGGCACCAAGCTGGAAATCAAG 903. PM08E11-HL
artificial nt
CAGGTGCAGCTGGTCCAGTCTGGCGCCGAAGTGATGAAGCCTGGCGCCTCCGTGA codon
optimized AGGTGTCCTGCAAGGCCTCCGGCTACACCATCACCGACACCTACATGGACTGGGT
GCGGCAGGCTCCTGGACAGGGCCTGGAATGGATCGCCCGGATCGACCCTGCCAA
CGGCGACTCCAAGTACGACCCTAAGTTCCAGGGCAGAGTGACCATGACCGCCGAC
ACCTCCACCAACACCGTGTACATGGAACTGTCCTCCCTGCGGTCTGAGGACACCGC
CGTGTACTACTGCGCCAGGGGCGGCATGATCTGGTACTTCGACGTGTGGGGCCAG
GGCACCACCGTGACAGTGTCCTCTGGCGGCGGAGGAAGTGGAGGTGGAGGATCT
GGTGGAGGCGGCTCCGAGATCGTGCTGACCCAGTCTCCTGCCACCATGTCTGTGT
CTCCCGGCGAGAGAGCCACCCTGTCCTGCTCCGCCTCCTCCTCCATCTCCTCCAAC
TACCTGCACTGGTATCAGCAGAAGCCTGGCCTGCCTCCTCGGCTGCTGATCTACCG
GACCTCCAACCTGGCCTCTGGCATCCCCGACAGGTTCTCCGGCTCTGGCTCCGGC
ACCGACTTCACCCTGACCATCTCCCGGCTGGAAGCTGAGGACTTCGCCACCTACTA
CTGCCAGCAGGGCTCCTCCCTGCCTTACACCTTCGGACAGGGCACCAAGCTGGAA ATCAAG 904.
PM08E11 HL .times. I2C artificial nt
CAGGTGCAGCTGGTCCAGTCTGGCGCCGAAGTGATGAAGCCTGGCGCCTCCGTGA HL codon
optimized AGGTGTCCTGCAAGGCCTCCGGCTACACCATCACCGACACCTACATGGACTGGGT
GCGGCAGGCTCCTGGACAGGGCCTGGAATGGATCGCCCGGATCGACCCTGCCAA
CGGCGACTCCAAGTACGACCCTAAGTTCCAGGGCAGAGTGACCATGACCGCCGAC
ACCTCCACCAACACCGTGTACATGGAACTGTCCTCCCTGCGGTCTGAGGACACCGC
CGTGTACTACTGCGCCAGGGGCGGCATGATCTGGTACTTCGACGTGTGGGGCCAG
GGCACCACCGTGACAGTGTCCTCTGGCGGCGGAGGAAGTGGAGGTGGAGGATCT
GGTGGAGGCGGCTCCGAGATCGTGCTGACCCAGTCTCCTGCCACCATGTCTGTGT
CTCCCGGCGAGAGAGCCACCCTGTCCTGCTCCGCCTCCTCCTCCATCTCCTCCAAC
TACCTGCACTGGTATCAGCAGAAGCCTGGCCTGCCTCCTCGGCTGCTGATCTACCG
GACCTCCAACCTGGCCTCTGGCATCCCCGACAGGTTCTCCGGCTCTGGCTCCGGC
ACCGACTTCACCCTGACCATCTCCCGGCTGGAAGCTGAGGACTTCGCCACCTACTA
CTGCCAGCAGGGCTCCTCCCTGCCTTACACCTTCGGACAGGGCACCAAGCTGGAA
ATCAAGTCTGGCGGAGGGGGCTCTGAAGTGCAGCTGGTGGAAAGCGGAGGGGGA
CTGGTGCAGCCCGGGGGAAGTCTGAAGCTGTCCTGTGCCGCCAGCGGCTTTACCT
TCAACAAGTACGCCATGAATTGGGTCCGACAGGCCCCAGGGAAAGGCCTGGAATG
GGTGGCACGGATCCGGTCCAAGTACAACAACTACGCCACCTACTACGCTGACTCCG
TGAAGGACAGATTCACCATCAGCCGGGACGACTCTAAGAACACCGCCTATCTGCAG
ATGAACAACCTGAAAACCGAGGATACAGCTGTGTACTATTGTGTGCGGCACGGCAA
CTTCGGCAACTCCTACATCTCCTACTGGGCCTATTGGGGACAGGGAACACTGGTCA
CCGTGTCTAGCGGAGGTGGCGGAAGTGGGGGAGGCGGATCTGGCGGTGGCGGAT
CCCAGACCGTGGTCACCCAGGAACCTTCCCTGACCGTCTCCCCAGGCGGCACCGT
GACCCTGACCTGTGGCTCCTCTACCGGCGCTGTGACCTCCGGCAACTACCCTAACT
GGGTGCAGCAGAAACCCGGACAGGCTCCTAGAGGCCTGATCGGCGGCACCAAGTT
TCTGGCCCCTGGCACCCCTGCCAGATTCTCCGGCTCCCTGCTGGGAGGCAAGGCC
GCTCTGACCCTGTCTGGCGTGCAGCCTGAGGACGAGGCCGAGTACTACTGTGTGC
TGTGGTACTCCAACAGATGGGTGTTCGGAGGCGGCACAAAGCTGACCGTGCTG 905. PM95H6-H
artificial aa
QVQLVQSGAEVMKPGASVKVSCKASGYNFKDTYMDWVKQTPEQGLEWMGRIDPANG
DSKYDPKFQGRVTITADTSTNTAYMELSSLRSEDTAVYYCARGGMIWYFDVWGQGTTV TVSS
906. PM95H6-HCDR1 artificial aa DTYMD 907. PM95H6-HCDR2 artificial
aa RIDPANGDSKYDPKFQG 908. PM95H6-HCDR3 artificial aa GGMIWYFDV 909.
PM95H6-H artificial nt
CAGGTGCAGCTGGTCCAGTCTGGGGCAGAGGTTATGAAGCCAGGGGCCTCAGTCA
AGGTGTCCTGCAAAGCTTCTGGCTACAACTTTAAAGACACCTATATGGACTGGGTGA
AGCAGACGCCTGAACAGGGCCTGGAATGGATGGGAAGGATTGATCCTGCGAATGG
TGATAGTAAATATGACCCGAAATTCCAGGGCAGGGTCACTATAACAGCAGACACAT
CCACCAACACAGCCTACATGGAGCTCAGCAGCCTGAGATCTGAGGACACTGCCGT
CTATTATTGTGCTAGAGGCGGGATGATATGGTACTTCGATGTCTGGGGCCAAGGGA
CCACGGTCACCGTCTCCTCA 910. PM95H6-L artificial aa
EIVLTQSPATLAVSPGEKVTLSCSASSSISSNYLHWYQQKPGLPPRLLIYRTSNLASGVP
DRFSGSGSGTDFTLTISRLEPEDFATYYCQQGSSLPYTFGQGTKLEIK 911. PM95H6-LCDR1
artificial aa SASSSISSNYLH 912. PM95H6-LCDR2 artificial aa RTSNLAS
913. PM95H6-LCDR3 artificial aa QQGSSLPYT 914. PM95H6-L artificial
nt GAGATCGTGCTCACCCAGTCTCCAGCCACCCTGGCTGTATCTCCCGGGGAGAAGG
TCACTCTCTCCTGCAGTGCCAGCTCAAGTATAAGTTCCAATTACTTGCATTGGTATC
AGCAGAAGCCAGGATTGCCCCCTAGACTCTTGATTTATAGGACATCCAATCTGGCTT
CTGGAGTCCCAGATCGCTTCAGTGGCAGTGGGTCTGGGACCGATTTCACTCTCACA
ATTAGCAGGCTGGAGCCTGAAGATTTTGCCACTTACTACTGCCAGCAGGGTAGTAG
TTTACCGTACACGTTCGGACAAGGGACCAAGCTTGAGATCAAA 915. PM95H6-HL
artificial aa
QVQLVQSGAEVMKPGASVKVSCKASGYNFKDTYMDWVKQTPEQGLEWMGRIDPANG
DSKYDPKFQGRVTITADTSTNTAYMELSSLRSEDTAVYYCARGGMIWYFDVWGQGTTV
TVSSGGGGSGGGGSGGGGSEIVLTQSPATLAVSPGEKVTLSCSASSSISSNYLHWYQ
QKPGLPPRLLIYRTSNLASGVPDRFSGSGSGTDFTLTISRLEPEDFATYYCQQGSSLPY
TFGQGTKLEIK 916. PM95H6-HL artificial nt
CAGGTGCAGCTGGTCCAGTCTGGGGCAGAGGTTATGAAGCCAGGGGCCTCAGTCA
AGGTGTCCTGCAAAGCTTCTGGCTACAACTTTAAAGACACCTATATGGACTGGGTGA
AGCAGACGCCTGAACAGGGCCTGGAATGGATGGGAAGGATTGATCCTGCGAATGG
TGATAGTAAATATGACCCGAAATTCCAGGGCAGGGTCACTATAACAGCAGACACAT
CCACCAACACAGCCTACATGGAGCTCAGCAGCCTGAGATCTGAGGACACTGCCGT
CTATTATTGTGCTAGAGGCGGGATGATATGGTACTTCGATGTCTGGGGCCAAGGGA
CCACGGTCACCGTCTCCTCAGGTGGTGGTGGTTCTGGCGGCGGCGGCTCCGGTG
GTGGTGGTTCTGAGATCGTGCTCACCCAGTCTCCAGCCACCCTGGCTGTATCTCCC
GGGGAGAAGGTCACTCTCTCCTGCAGTGCCAGCTCAAGTATAAGTTCCAATTACTT
GCATTGGTATCAGCAGAAGCCAGGATTGCCCCCTAGACTCTTGATTTATAGGACATC
CAATCTGGCTTCTGGAGTCCCAGATCGCTTCAGTGGCAGTGGGTCTGGGACCGATT
TCACTCTCACAATTAGCAGGCTGGAGCCTGAAGATTTTGCCACTTACTACTGCCAGC
AGGGTAGTAGTTTACCGTACACGTTCGGACAAGGGACCAAGCTTGAGATCAAA 917. PM95H6
HL .times. I2C artificial aa
QVQLVQSGAEVMKPGASVKVSCKASGYNFKDTYMDWVKQTPEQGLEWMGRIDPANG HL
DSKYDPKFQGRVTITADTSTNTAYMELSSLRSEDTAVYYCARGGMIWYFDVWGQGTTV
TVSSGGGGSGGGGSGGGGSEIVLTQSPATLAVSPGEKVTLSCSASSSISSNYLHWYQ
QKPGLPPRLLIYRTSNLASGVPDRFSGSGSGTDFTLTISRLEPEDFATYYCQQGSSLPY
TFGQGTKLEIKSGGGGSEVQLVESGGGLVQPGGSLKLSCAASGFTFNKYAMNWVRQA
PGKGLEWVARIRSKYNNYATYYADSVKDRFTISRDDSKNTAYLQMNNLKTEDTAVYYC
VRHGNFGNSYISYWAYWGQGTLVTVSSGGGGSGGGGSGGGGSQTVVTQEPSLTVSP
GGTVTLTCGSSTGAVTSGNYPNWVQQKPGQAPRGLIGGTKFLAPGTPARFSGSLLGG
KAALTLSGVQPEDEAEYYCVLWYSNRWVFGGGTKLTVL 918. PM95H6 HL .times. I2C
artificial nt
CAGGTGCAGCTGGTCCAGTCTGGGGCAGAGGTTATGAAGCCAGGGGCCTCAGTCA HL
AGGTGTCCTGCAAAGCTTCTGGCTACAACTTTAAAGACACCTATATGGACTGGGTGA
AGCAGACGCCTGAACAGGGCCTGGAATGGATGGGAAGGATTGATCCTGCGAATGG
TGATAGTAAATATGACCCGAAATTCCAGGGCAGGGTCACTATAACAGCAGACACAT
CCACCAACACAGCCTACATGGAGCTCAGCAGCCTGAGATCTGAGGACACTGCCGT
CTATTATTGTGCTAGAGGCGGGATGATATGGTACTTCGATGTCTGGGGCCAAGGGA
CCACGGTCACCGTCTCCTCAGGTGGTGGTGGTTCTGGCGGCGGCGGCTCCGGTG
GTGGTGGTTCTGAGATCGTGCTCACCCAGTCTCCAGCCACCCTGGCTGTATCTCCC
GGGGAGAAGGTCACTCTCTCCTGCAGTGCCAGCTCAAGTATAAGTTCCAATTACTT
GCATTGGTATCAGCAGAAGCCAGGATTGCCCCCTAGACTCTTGATTTATAGGACATC
CAATCTGGCTTCTGGAGTCCCAGATCGCTTCAGTGGCAGTGGGTCTGGGACCGATT
TCACTCTCACAATTAGCAGGCTGGAGCCTGAAGATTTTGCCACTTACTACTGCCAGC
AGGGTAGTAGTTTACCGTACACGTTCGGACAAGGGACCAAGCTTGAGATCAAATCC
GGAGGTGGTGGATCCGAGGTGCAGCTGGTCGAGTCTGGAGGAGGATTGGTGCAG
CCTGGAGGGTCATTGAAACTCTCATGTGCAGCCTCTGGATTCACCTTCAATAAGTAC
GCCATGAACTGGGTCCGCCAGGCTCCAGGAAAGGGTTTGGAATGGGTTGCTCGCA
TAAGAAGTAAATATAATAATTATGCAACATATTATGCCGATTCAGTGAAAGACAGGTT
CACCATCTCCAGAGATGATTCAAAAAACACTGCCTATCTACAAATGAACAACTTGAA
AACTGAGGACACTGCCGTGTACTACTGTGTGAGACATGGGAACTTCGGTAATAGCT
ACATATCCTACTGGGCTTACTGGGGCCAAGGGACTCTGGTCACCGTCTCCTCAGGT
GGTGGTGGTTCTGGCGGCGGCGGCTCCGGTGGTGGTGGTTCTCAGACTGTTGTGA
CTCAGGAACCTTCACTCACCGTATCACCTGGTGGAACAGTCACACTCACTTGTGGC
TCCTCGACTGGGGCTGTTACATCTGGCAACTACCCAAACTGGGTCCAACAAAAACC
AGGTCAGGCACCCCGTGGTCTAATAGGTGGGACTAAGTTCCTCGCCCCCGGTACT
CCTGCCAGATTCTCAGGCTCCCTGCTTGGAGGCAAGGCTGCCCTCACCCTCTCAG
GGGTACAGCCAGAGGATGAGGCAGAATATTACTGTGTTCTATGGTACAGCAACCGC
TGGGTGTTCGGTGGAGGAACCAAACTGACTGTCCTA 919. PM95H6-H artificial nt
CAGGTGCAGCTGGTCCAGTCTGGCGCCGAAGTGATGAAGCCTGGCGCCTCCGTGA codon
optimized AGGTGTCCTGCAAGGCCTCCGGCTACAACTTCAAGGACACCTACATGGACTGGGTG
AAACAGACCCCTGAGCAGGGCCTGGAATGGATGGGCCGGATCGACCCTGCCAACG
GCGACTCCAAGTACGACCCTAAGTTCCAGGGCAGAGTGACCATCACCGCCGACAC
CTCCACCAACACCGCCTACATGGAACTGTCCTCCCTGCGGTCTGAGGACACCGCC
GTGTACTACTGCGCCAGGGGCGGCATGATCTGGTACTTCGACGTGTGGGGCCAGG
GCACCACCGTGACAGTGTCCTCT 920. PM95H6-L artificial nt
GAGATCGTGCTGACCCAGTCTCCTGCCACCCTGGCTGTGTCTCCCGGCGAGAAAG codon
optimized TGACCCTGTCCTGCTCCGCCTCCTCCTCCATCTCCTCCAACTACCTGCACTGGTATC
AGCAGAAGCCTGGCCTGCCTCCTCGGCTGCTGATCTACCGGACCTCCAACCTGGC
CTCTGGCGTGCCCGACAGGTTCTCCGGCTCTGGCTCCGGCACCGACTTCACCCTG
ACCATCTCCCGGCTGGAACCTGAGGACTTCGCCACCTACTACTGCCAGCAGGGCT
CCTCCCTGCCTTACACCTTCGGACAGGGCACCAAGCTGGAAATCAAG 921. PM95H6-HL
artificial nt
CAGGTGCAGCTGGTCCAGTCTGGCGCCGAAGTGATGAAGCCTGGCGCCTCCGTGA codon
optimized AGGTGTCCTGCAAGGCCTCCGGCTACAACTTCAAGGACACCTACATGGACTGGGTG
AAACAGACCCCTGAGCAGGGCCTGGAATGGATGGGCCGGATCGACCCTGCCAACG
GCGACTCCAAGTACGACCCTAAGTTCCAGGGCAGAGTGACCATCACCGCCGACAC
CTCCACCAACACCGCCTACATGGAACTGTCCTCCCTGCGGTCTGAGGACACCGCC
GTGTACTACTGCGCCAGGGGCGGCATGATCTGGTACTTCGACGTGTGGGGCCAGG
GCACCACCGTGACAGTGTCCTCTGGCGGCGGAGGAAGTGGAGGTGGAGGATCTG
GTGGAGGCGGCTCCGAGATCGTGCTGACCCAGTCTCCTGCCACCCTGGCTGTGTC
TCCCGGCGAGAAAGTGACCCTGTCCTGCTCCGCCTCCTCCTCCATCTCCTCCAACT
ACCTGCACTGGTATCAGCAGAAGCCTGGCCTGCCTCCTCGGCTGCTGATCTACCG
GACCTCCAACCTGGCCTCTGGCGTGCCCGACAGGTTCTCCGGCTCTGGCTCCGGC
ACCGACTTCACCCTGACCATCTCCCGGCTGGAACCTGAGGACTTCGCCACCTACTA
CTGCCAGCAGGGCTCCTCCCTGCCTTACACCTTCGGACAGGGCACCAAGCTGGAA ATCAAG 922.
PM95H6 HL .times. I2C artificial nt
CAGGTGCAGCTGGTCCAGTCTGGCGCCGAAGTGATGAAGCCTGGCGCCTCCGTGA HL codon
optimized AGGTGTCCTGCAAGGCCTCCGGCTACAACTTCAAGGACACCTACATGGACTGGGTG
AAACAGACCCCTGAGCAGGGCCTGGAATGGATGGGCCGGATCGACCCTGCCAACG
GCGACTCCAAGTACGACCCTAAGTTCCAGGGCAGAGTGACCATCACCGCCGACAC
CTCCACCAACACCGCCTACATGGAACTGTCCTCCCTGCGGTCTGAGGACACCGCC
GTGTACTACTGCGCCAGGGGCGGCATGATCTGGTACTTCGACGTGTGGGGCCAGG
GCACCACCGTGACAGTGTCCTCTGGCGGCGGAGGAAGTGGAGGTGGAGGATCTG
GTGGAGGCGGCTCCGAGATCGTGCTGACCCAGTCTCCTGCCACCCTGGCTGTGTC
TCCCGGCGAGAAAGTGACCCTGTCCTGCTCCGCCTCCTCCTCCATCTCCTCCAACT
ACCTGCACTGGTATCAGCAGAAGCCTGGCCTGCCTCCTCGGCTGCTGATCTACCG
GACCTCCAACCTGGCCTCTGGCGTGCCCGACAGGTTCTCCGGCTCTGGCTCCGGC
ACCGACTTCACCCTGACCATCTCCCGGCTGGAACCTGAGGACTTCGCCACCTACTA
CTGCCAGCAGGGCTCCTCCCTGCCTTACACCTTCGGACAGGGCACCAAGCTGGAA
ATCAAGTCTGGCGGAGGGGGCTCTGAAGTGCAGCTGGTGGAAAGCGGAGGGGGA
CTGGTGCAGCCCGGGGGAAGTCTGAAGCTGTCCTGTGCCGCCAGCGGCTTTACCT
TCAACAAGTACGCCATGAATTGGGTCCGACAGGCCCCAGGGAAAGGCCTGGAATG
GGTGGCACGGATCCGGTCCAAGTACAACAACTACGCCACCTACTACGCTGACTCCG
TGAAGGACAGATTCACCATCAGCCGGGACGACTCTAAGAACACCGCCTATCTGCAG
ATGAACAACCTGAAAACCGAGGATACAGCTGTGTACTATTGTGTGCGGCACGGCAA
CTTCGGCAACTCCTACATCTCCTACTGGGCCTATTGGGGACAGGGAACACTGGTCA
CCGTGTCTAGCGGAGGTGGCGGAAGTGGGGGAGGCGGATCTGGCGGTGGCGGAT
CCCAGACCGTGGTCACCCAGGAACCTTCCCTGACCGTCTCCCCAGGCGGCACCGT
GACCCTGACCTGTGGCTCCTCTACCGGCGCTGTGACCTCCGGCAACTACCCTAACT
GGGTGCAGCAGAAACCCGGACAGGCTCCTAGAGGCCTGATCGGCGGCACCAAGTT
TCTGGCCCCTGGCACCCCTGCCAGATTCTCCGGCTCCCTGCTGGGAGGCAAGGCC
GCTCTGACCCTGTCTGGCGTGCAGCCTGAGGACGAGGCCGAGTACTACTGTGTGC
TGTGGTACTCCAACAGATGGGTGTTCGGAGGCGGCACAAAGCTGACCGTGCTG 923. PM95A8-H
artificial aa
QVQLVQSGAEVMKPGASVKVSCKASGYNFKDTYMDWVKQTPEQGLEWMGRIDPANG
DSKYDPKFQGRVTITADTSTNTAYMELSSLRSEDTAVYYCARGGMIWYFDVWGQGTTV TVSS
924. PM95A8-HCDR1 artificial aa DTYMD 925. PM95A8-HCDR2 artificial
aa RIDPANGDSKYDPKFQG 926. PM95A8-HCDR3 artificial aa GGMIWYFDV 927.
PM95A8-H artificial nt
CAGGTGCAGCTGGTCCAGTCTGGGGCAGAGGTTATGAAGCCAGGGGCCTCAGTCA
AGGTGTCCTGCAAAGCTTCTGGCTACAACTTTAAAGACACCTATATGGACTGGGTGA
AGCAGACGCCTGAACAGGGCCTGGAATGGATGGGAAGGATTGATCCTGCGAATGG
TGATAGTAAATATGACCCGAAATTCCAGGGCAGGGTCACTATAACAGCAGACACAT
CCACCAACACAGCCTACATGGAGCTCAGCAGCCTGAGATCTGAGGACACTGCCGT
CTATTATTGTGCTAGAGGCGGGATGATATGGTACTTCGATGTCTGGGGCCAAGGGA
CCACGGTCACCGTCTCCTCA 928. PM95A8-L artificial aa
EIVLTQSPATMSVSPGERATLSCSASSSISSNYLHWYQQKPGLPPRLLIYRTSNLASGIP
DRFSGSGSGTDFTLTISRLEAEDFATYYCQQGSSLPYTFGQGTKLEIK 929. PM95A8-LCDR1
artificial aa SASSSISSNYLH 930. PM95A8-LCDR2 artificial aa RTSNLAS
931. PM95A8-LCDR3 artificial aa QQGSSLPYT 932. PM95A8-L artificial
nt GAGATCGTGCTCACCCAGTCTCCAGCCACCATGTCTGTATCTCCCGGGGAGAGGG
CCACTCTCTCCTGCAGTGCCAGCTCAAGTATAAGTTCCAATTACTTGCATTGGTATC
AGCAGAAGCCAGGATTGCCCCCTAGACTCTTGATTTATAGGACATCCAATCTGGCTT
CTGGAATCCCAGATCGCTTCAGTGGCAGTGGGTCTGGGACCGATTTCACTCTCACA
ATTAGCAGGCTGGAGGCTGAAGATTTTGCCACTTACTACTGCCAGCAGGGTAGTAG
TTTACCGTACACGTTCGGACAAGGGACCAAGCTTGAGATCAAA 933. PM95A8-HL
artificial aa
QVQLVQSGAEVMKPGASVKVSCKASGYNFKDTYMDWVKQTPEQGLEWMGRIDPANG
DSKYDPKFQGRVTITADTSTNTAYMELSSLRSEDTAVYYCARGGMIWYFDVWGQGTTV
TVSSGGGGSGGGGSGGGGSEIVLTQSPATMSVSPGERATLSCSASSSISSNYLHWYQ
QKPGLPPRLLIYRTSNLASGIPDRFSGSGSGTDFTLTISRLEAEDFATYYCQQGSSLPYT
FGQGTKLEIK 934. PM95A8-HL artificial nt
CAGGTGCAGCTGGTCCAGTCTGGGGCAGAGGTTATGAAGCCAGGGGCCTCAGTCA
AGGTGTCCTGCAAAGCTTCTGGCTACAACTTTAAAGACACCTATATGGACTGGGTGA
AGCAGACGCCTGAACAGGGCCTGGAATGGATGGGAAGGATTGATCCTGCGAATGG
TGATAGTAAATATGACCCGAAATTCCAGGGCAGGGTCACTATAACAGCAGACACAT
CCACCAACACAGCCTACATGGAGCTCAGCAGCCTGAGATCTGAGGACACTGCCGT
CTATTATTGTGCTAGAGGCGGGATGATATGGTACTTCGATGTCTGGGGCCAAGGGA
CCACGGTCACCGTCTCCTCAGGTGGTGGTGGTTCTGGCGGCGGCGGCTCCGGTG
GTGGTGGTTCTGAGATCGTGCTCACCCAGTCTCCAGCCACCATGTCTGTATCTCCC
GGGGAGAGGGCCACTCTCTCCTGCAGTGCCAGCTCAAGTATAAGTTCCAATTACTT
GCATTGGTATCAGCAGAAGCCAGGATTGCCCCCTAGACTCTTGATTTATAGGACATC
CAATCTGGCTTCTGGAATCCCAGATCGCTTCAGTGGCAGTGGGTCTGGGACCGATT
TCACTCTCACAATTAGCAGGCTGGAGGCTGAAGATTTTGCCACTTACTACTGCCAGC
AGGGTAGTAGTTTACCGTACACGTTCGGACAAGGGACCAAGCTTGAGATCAAA 935. PM95A8
HL .times. I2C artificial aa
QVQLVQSGAEVMKPGASVKVSCKASGYNFKDTYMDWVKQTPEQGLEWMGRIDPANG HL
DSKYDPKFQGRVTITADTSTNTAYMELSSLRSEDTAVYYCARGGMIWYFDVWGQGTTV
TVSSGGGGSGGGGSGGGGSEIVLTQSPATMSVSPGERATLSCSASSSISSNYLHWYQ
QKPGLPPRLLIYRTSNLASGIPDRFSGSGSGTDFTLTISRLEAEDFATYYCQQGSSLPYT
FGQGTKLEIKSGGGGSEVQLVESGGGLVQPGGSLKLSCAASGFTFNKYAMNWVRQAP
GKGLEWVARIRSKYNNYATYYADSVKDRFTISRDDSKNTAYLQMNNLKTEDTAVYYCV
RHGNFGNSYISYWAYWGQGTLVTVSSGGGGSGGGGSGGGGSQTVVTQEPSLTVSPG
GTVTLTCGSSTGAVTSGNYPNWVQQKPGQAPRGLIGGTKFLAPGTPARFSGSLLGGK
AALTLSGVQPEDEAEYYCVLWYSNRWVFGGGTKLTVL 936. PM95A8 HL .times. I2C
artificial nt
CAGGTGCAGCTGGTCCAGTCTGGGGCAGAGGTTATGAAGCCAGGGGCCTCAGTCA HL
AGGTGTCCTGCAAAGCTTCTGGCTACAACTTTAAAGACACCTATATGGACTGGGTGA
AGCAGACGCCTGAACAGGGCCTGGAATGGATGGGAAGGATTGATCCTGCGAATGG
TGATAGTAAATATGACCCGAAATTCCAGGGCAGGGTCACTATAACAGCAGACACAT
CCACCAACACAGCCTACATGGAGCTCAGCAGCCTGAGATCTGAGGACACTGCCGT
CTATTATTGTGCTAGAGGCGGGATGATATGGTACTTCGATGTCTGGGGCCAAGGGA
CCACGGTCACCGTCTCCTCAGGTGGTGGTGGTTCTGGCGGCGGCGGCTCCGGTG
GTGGTGGTTCTGAGATCGTGCTCACCCAGTCTCCAGCCACCATGTCTGTATCTCCC
GGGGAGAGGGCCACTCTCTCCTGCAGTGCCAGCTCAAGTATAAGTTCCAATTACTT
GCATTGGTATCAGCAGAAGCCAGGATTGCCCCCTAGACTCTTGATTTATAGGACATC
CAATCTGGCTTCTGGAATCCCAGATCGCTTCAGTGGCAGTGGGTCTGGGACCGATT
TCACTCTCACAATTAGCAGGCTGGAGGCTGAAGATTTTGCCACTTACTACTGCCAGC
AGGGTAGTAGTTTACCGTACACGTTCGGACAAGGGACCAAGCTTGAGATCAAATCC
GGAGGTGGTGGATCCGAGGTGCAGCTGGTCGAGTCTGGAGGAGGATTGGTGCAG
CCTGGAGGGTCATTGAAACTCTCATGTGCAGCCTCTGGATTCACCTTCAATAAGTAC
GCCATGAACTGGGTCCGCCAGGCTCCAGGAAAGGGTTTGGAATGGGTTGCTCGCA
TAAGAAGTAAATATAATAATTATGCAACATATTATGCCGATTCAGTGAAAGACAGGTT
CACCATCTCCAGAGATGATTCAAAAAACACTGCCTATCTACAAATGAACAACTTGAA
AACTGAGGACACTGCCGTGTACTACTGTGTGAGACATGGGAACTTCGGTAATAGCT
ACATATCCTACTGGGCTTACTGGGGCCAAGGGACTCTGGTCACCGTCTCCTCAGGT
GGTGGTGGTTCTGGCGGCGGCGGCTCCGGTGGTGGTGGTTCTCAGACTGTTGTGA
CTCAGGAACCTTCACTCACCGTATCACCTGGTGGAACAGTCACACTCACTTGTGGC
TCCTCGACTGGGGCTGTTACATCTGGCAACTACCCAAACTGGGTCCAACAAAAACC
AGGTCAGGCACCCCGTGGTCTAATAGGTGGGACTAAGTTCCTCGCCCCCGGTACT
CCTGCCAGATTCTCAGGCTCCCTGCTTGGAGGCAAGGCTGCCCTCACCCTCTCAG
GGGTACAGCCAGAGGATGAGGCAGAATATTACTGTGTTCTATGGTACAGCAACCGC
TGGGTGTTCGGTGGAGGAACCAAACTGACTGTCCTA 937. PM95A8-H artificial nt
CAGGTGCAGCTGGTCCAGTCTGGCGCCGAAGTGATGAAGCCTGGCGCCTCCGTGA codon
optimized AGGTGTCCTGCAAGGCCTCCGGCTACAACTTCAAGGACACCTACATGGACTGGGTG
AAACAGACCCCTGAGCAGGGCCTGGAATGGATGGGCCGGATCGACCCTGCCAACG
GCGACTCCAAGTACGACCCTAAGTTCCAGGGCAGAGTGACCATCACCGCCGACAC
CTCCACCAACACCGCCTACATGGAACTGTCCTCCCTGCGGTCTGAGGACACCGCC
GTGTACTACTGCGCCAGGGGCGGCATGATCTGGTACTTCGACGTGTGGGGCCAGG
GCACCACCGTGACAGTGTCCTCT 938. PM95A8-HCDR1 artificial nt
GAGATCGTGCTGACCCAGTCTCCTGCCACCATGTCCGTGTCTCCCGGCGAGAGGG codon
optimized CTACCCTGTCCTGCTCCGCCTCCTCCTCCATCTCCTCCAACTACCTGCACTGGTATC
AGCAGAAGCCTGGCCTGCCTCCTCGGCTGCTGATCTACCGGACCTCCAACCTGGC
CTCTGGCATCCCCGACAGGTTCTCCGGCTCTGGCTCCGGCACCGACTTCACCCTG
ACCATCTCCCGGCTGGAAGCTGAGGACTTCGCCACCTACTACTGCCAGCAGGGCT
CCTCCCTGCCTTACACCTTCGGACAGGGCACCAAGCTGGAAATCAAG 939. PM95A8-HCDR2
artificial nt
CAGGTGCAGCTGGTCCAGTCTGGCGCCGAAGTGATGAAGCCTGGCGCCTCCGTGA codon
optimized AGGTGTCCTGCAAGGCCTCCGGCTACAACTTCAAGGACACCTACATGGACTGGGTG
AAACAGACCCCTGAGCAGGGCCTGGAATGGATGGGCCGGATCGACCCTGCCAACG
GCGACTCCAAGTACGACCCTAAGTTCCAGGGCAGAGTGACCATCACCGCCGACAC
CTCCACCAACACCGCCTACATGGAACTGTCCTCCCTGCGGTCTGAGGACACCGCC
GTGTACTACTGCGCCAGGGGCGGCATGATCTGGTACTTCGACGTGTGGGGCCAGG
GCACCACCGTGACAGTGTCCTCTGGCGGCGGAGGAAGTGGAGGTGGAGGATCTG
GTGGAGGCGGCTCCGAGATCGTGCTGACCCAGTCTCCTGCCACCATGTCCGTGTC
TCCCGGCGAGAGGGCTACCCTGTCCTGCTCCGCCTCCTCCTCCATCTCCTCCAACT
ACCTGCACTGGTATCAGCAGAAGCCTGGCCTGCCTCCTCGGCTGCTGATCTACCG
GACCTCCAACCTGGCCTCTGGCATCCCCGACAGGTTCTCCGGCTCTGGCTCCGGC
ACCGACTTCACCCTGACCATCTCCCGGCTGGAAGCTGAGGACTTCGCCACCTACTA
CTGCCAGCAGGGCTCCTCCCTGCCTTACACCTTCGGACAGGGCACCAAGCTGGAA ATCAAG 940.
PM95A8 HL .times. I2C artificial nt
CAGGTGCAGCTGGTCCAGTCTGGCGCCGAAGTGATGAAGCCTGGCGCCTCCGTGA HL_codon
optimized AGGTGTCCTGCAAGGCCTCCGGCTACAACTTCAAGGACACCTACATGGACTGGGTG
AAACAGACCCCTGAGCAGGGCCTGGAATGGATGGGCCGGATCGACCCTGCCAACG
GCGACTCCAAGTACGACCCTAAGTTCCAGGGCAGAGTGACCATCACCGCCGACAC
CTCCACCAACACCGCCTACATGGAACTGTCCTCCCTGCGGTCTGAGGACACCGCC
GTGTACTACTGCGCCAGGGGCGGCATGATCTGGTACTTCGACGTGTGGGGCCAGG
GCACCACCGTGACAGTGTCCTCTGGCGGCGGAGGAAGTGGAGGTGGAGGATCTG
GTGGAGGCGGCTCCGAGATCGTGCTGACCCAGTCTCCTGCCACCATGTCCGTGTC
TCCCGGCGAGAGGGCTACCCTGTCCTGCTCCGCCTCCTCCTCCATCTCCTCCAACT
ACCTGCACTGGTATCAGCAGAAGCCTGGCCTGCCTCCTCGGCTGCTGATCTACCG
GACCTCCAACCTGGCCTCTGGCATCCCCGACAGGTTCTCCGGCTCTGGCTCCGGC
ACCGACTTCACCCTGACCATCTCCCGGCTGGAAGCTGAGGACTTCGCCACCTACTA
CTGCCAGCAGGGCTCCTCCCTGCCTTACACCTTCGGACAGGGCACCAAGCTGGAA
ATCAAGTCTGGCGGAGGGGGCTCTGAAGTGCAGCTGGTGGAAAGCGGAGGGGGA
CTGGTGCAGCCCGGGGGAAGTCTGAAGCTGTCCTGTGCCGCCAGCGGCTTTACCT
TCAACAAGTACGCCATGAATTGGGTCCGACAGGCCCCAGGGAAAGGCCTGGAATG
GGTGGCACGGATCCGGTCCAAGTACAACAACTACGCCACCTACTACGCTGACTCCG
TGAAGGACAGATTCACCATCAGCCGGGACGACTCTAAGAACACCGCCTATCTGCAG
ATGAACAACCTGAAAACCGAGGATACAGCTGTGTACTATTGTGTGCGGCACGGCAA
CTTCGGCAACTCCTACATCTCCTACTGGGCCTATTGGGGACAGGGAACACTGGTCA
CCGTGTCTAGCGGAGGTGGCGGAAGTGGGGGAGGCGGATCTGGCGGTGGCGGAT
CCCAGACCGTGGTCACCCAGGAACCTTCCCTGACCGTCTCCCCAGGCGGCACCGT
GACCCTGACCTGTGGCTCCTCTACCGGCGCTGTGACCTCCGGCAACTACCCTAACT
GGGTGCAGCAGAAACCCGGACAGGCTCCTAGAGGCCTGATCGGCGGCACCAAGTT
TCTGGCCCCTGGCACCCCTGCCAGATTCTCCGGCTCCCTGCTGGGAGGCAAGGCC
GCTCTGACCCTGTCTGGCGTGCAGCCTGAGGACGAGGCCGAGTACTACTGTGTGC
TGTGGTACTCCAACAGATGGGTGTTCGGAGGCGGCACAAAGCTGACCGTGCTG 941.
PM07A12-H artificial aa
QVQLVESGGGLVKPGGSLRLSCAASGFTFSDYYMSWVRQAPGKGLEWVASISDGGSN
TYYSDIIKGRFTISRDNAKNNLYLQMNSLRAEDTAVYYCARGFPLLRHGAFDYWGQGTL VTVSS
942. PM07A12-HCDR1 artificial aa DYYMS 943. PM07A12-HCDR2
artificial aa SISDGGSNTYYSDIIKG 944. PM07A12-HCDR3 artificial aa
GFPLLRHGAFDY 945. PM07A12-H artificial nt
CAGGTGCAGCTGGTCGAGTCTGGCGGCGGACTGGTGAAGCCTGGCGGGTCCCTG
AGGCTGTCCTGTGCCGCCTCCGGCTTCACCTTCTCCGACTACTACATGAGCTGGGT
CCGCCAGGCCCCTGGGAAGGGGCTGGAATGGGTGGCCTCCATCTCCGACGGCGG
CTCCAACACCTACTACTCCGACATCATCAAGGGCCGGTTCACCATCTCCCGGGACA
ACGCCAAGAACAATCTGTACCTGCAGATGAACTCCCTGAGGGCCGAGGACACCGC
CGTGTACTACTGCGCCCGGGGCTTCCCTCTGCTGAGACACGGCGCCTTCGATTACT
GGGGCCAGGGCACCCTGGTCACCGTCTCCTCA 946. PM07A12-L artificial aa
DIQMTQSPSSLSASVGDRVTITCRASQNVDTNVAWYQQKPGQAPKSLIYSATYRYSDV
PSRFSGSASGTDFTLTISSVQSEDFATYYCQQYNSYPYTFGGGTKLEIK 947.
PM07A12-LCDR1 artificial aa RASQNVDTNVA 948. PM07A12-LCDR2
artificial aa SATYRYS 949. PM07A12-LCDR3 artificial aa QQYNSYPYT
950. PM07A12-L artificial nt
GACATCCAGATGACCCAGTCCCCCAGCTCCCTGTCCGCCTCCGTGGGCGACAGAG
TGACCATCACCTGCAGGGCCTCCCAGAACGTGGACACCAACGTGGCCTGGTATCA
GCAGAAGCCCGGCCAGGCCCCTAAGTCCCTGATCTACTCCGCCACCTACCGGTAC
TCTGACGTGCCTTCCCGGTTCTCCGGCTCCGCGTCCGGCACCGACTTCACCCTGA
CCATCTCCAGCGTGCAGTCTGAGGACTTCGCCACGTACTACTGCCAGCAGTACAAC
TCCTACCCTTACACCTTCGGCGGAGGGACCAAGCTGGAAATCAAG 951. PM07A12-HL
artificial aa
QVQLVESGGGLVKPGGSLRLSCAASGFTFSDYYMSWVRQAPGKGLEWVASISDGGSN
TYYSDIIKGRFTISRDNAKNNLYLQMNSLRAEDTAVYYCARGFPLLRHGAFDYWGQGTL
VTVSSGGGGSGGGGSGGGGSDIQMTQSPSSLSASVGDRVTITCRASQNVDTNVAWY
QQKPGQAPKSLIYSATYRYSDVPSRFSGSASGTDFTLTISSVQSEDFATYYCQQYNSYP
YTFGGGTKLEIK 952. PM07A12-HL artificial nt
CAGGTGCAGCTGGTCGAGTCTGGCGGCGGACTGGTGAAGCCTGGCGGGTCCCTG
AGGCTGTCCTGTGCCGCCTCCGGCTTCACCTTCTCCGACTACTACATGAGCTGGGT
CCGCCAGGCCCCTGGGAAGGGGCTGGAATGGGTGGCCTCCATCTCCGACGGCGG
CTCCAACACCTACTACTCCGACATCATCAAGGGCCGGTTCACCATCTCCCGGGACA
ACGCCAAGAACAATCTGTACCTGCAGATGAACTCCCTGAGGGCCGAGGACACCGC
CGTGTACTACTGCGCCCGGGGCTTCCCTCTGCTGAGACACGGCGCCTTCGATTACT
GGGGCCAGGGCACCCTGGTCACCGTCTCCTCAGGTGGTGGTGGTTCTGGCGGCG
GCGGCTCCGGTGGTGGTGGTTCTGACATCCAGATGACCCAGTCCCCCAGCTCCCT
GTCCGCCTCCGTGGGCGACAGAGTGACCATCACCTGCAGGGCCTCCCAGAACGTG
GACACCAACGTGGCCTGGTATCAGCAGAAGCCCGGCCAGGCCCCTAAGTCCCTGA
TCTACTCCGCCACCTACCGGTACTCTGACGTGCCTTCCCGGTTCTCCGGCTCCGCG
TCCGGCACCGACTTCACCCTGACCATCTCCAGCGTGCAGTCTGAGGACTTCGCCAC
GTACTACTGCCAGCAGTACAACTCCTACCCTTACACCTTCGGCGGAGGGACCAAGC
TGGAAATCAAG 953. PM07A12 HL .times. I2C artificial aa
QVQLVESGGGLVKPGGSLRLSCAASGFTFSDYYMSWVRQAPGKGLEWVASISDGGSN HL
TYYSDIIKGRFTISRDNAKNNLYLQMNSLRAEDTAVYYCARGFPLLRHGAFDYWGQGTL
VTVSSGGGGSGGGGSGGGGSDIQMTQSPSSLSASVGDRVTITCRASQNVDTNVAWY
QQKPGQAPKSLIYSATYRYSDVPSRFSGSASGTDFTLTISSVQSEDFATYYCQQYNSYP
YTFGGGTKLEIKSGGGGSEVQLVESGGGLVQPGGSLKLSCAASGFTFNKYAMNWVRQ
APGKGLEWVARIRSKYNNYATYYADSVKDRFTISRDDSKNTAYLQMNNLKTEDTAVYY
CVRHGNFGNSYISYWAYWGQGTLVTVSSGGGGSGGGGSGGGGSQTVVTQEPSLTVS
PGGTVTLTCGSSTGAVTSGNYPNWVQQKPGQAPRGLIGGTKFLAPGTPARFSGSLLG
GKAALTLSGVQPEDEAEYYCVLWYSNRWVFGGGTKLTVL 954. PM07A12 HL .times. I2C
artificial nt
CAGGTGCAGCTGGTCGAGTCTGGCGGCGGACTGGTGAAGCCTGGCGGGTCCCTG HL
AGGCTGTCCTGTGCCGCCTCCGGCTTCACCTTCTCCGACTACTACATGAGCTGGGT
CCGCCAGGCCCCTGGGAAGGGGCTGGAATGGGTGGCCTCCATCTCCGACGGCGG
CTCCAACACCTACTACTCCGACATCATCAAGGGCCGGTTCACCATCTCCCGGGACA
ACGCCAAGAACAATCTGTACCTGCAGATGAACTCCCTGAGGGCCGAGGACACCGC
CGTGTACTACTGCGCCCGGGGCTTCCCTCTGCTGAGACACGGCGCCTTCGATTACT
GGGGCCAGGGCACCCTGGTCACCGTCTCCTCAGGTGGTGGTGGTTCTGGCGGCG
GCGGCTCCGGTGGTGGTGGTTCTGACATCCAGATGACCCAGTCCCCCAGCTCCCT
GTCCGCCTCCGTGGGCGACAGAGTGACCATCACCTGCAGGGCCTCCCAGAACGTG
GACACCAACGTGGCCTGGTATCAGCAGAAGCCCGGCCAGGCCCCTAAGTCCCTGA
TCTACTCCGCCACCTACCGGTACTCTGACGTGCCTTCCCGGTTCTCCGGCTCCGCG
TCCGGCACCGACTTCACCCTGACCATCTCCAGCGTGCAGTCTGAGGACTTCGCCAC
GTACTACTGCCAGCAGTACAACTCCTACCCTTACACCTTCGGCGGAGGGACCAAGC
TGGAAATCAAGTCCGGAGGTGGTGGATCCGAGGTGCAGCTGGTCGAGTCTGGAGG
AGGATTGGTGCAGCCTGGAGGGTCATTGAAACTCTCATGTGCAGCCTCTGGATTCA
CCTTCAATAAGTACGCCATGAACTGGGTCCGCCAGGCTCCAGGAAAGGGTTTGGAA
TGGGTTGCTCGCATAAGAAGTAAATATAATAATTATGCAACATATTATGCCGATTCAG
TGAAAGACAGGTTCACCATCTCCAGAGATGATTCAAAAAACACTGCCTATCTACAAA
TGAACAACTTGAAAACTGAGGACACTGCCGTGTACTACTGTGTGAGACATGGGAAC
TTCGGTAATAGCTACATATCCTACTGGGCTTACTGGGGCCAAGGGACTCTGGTCAC
CGTCTCCTCAGGTGGTGGTGGTTCTGGCGGCGGCGGCTCCGGTGGTGGTGGTTCT
CAGACTGTTGTGACTCAGGAACCTTCACTCACCGTATCACCTGGTGGAACAGTCAC
ACTCACTTGTGGCTCCTCGACTGGGGCTGTTACATCTGGCAACTACCCAAACTGGG
TCCAACAAAAACCAGGTCAGGCACCCCGTGGTCTAATAGGTGGGACTAAGTTCCTC
GCCCCCGGTACTCCTGCCAGATTCTCAGGCTCCCTGCTTGGAGGCAAGGCTGCCC
TCACCCTCTCAGGGGTACAGCCAGAGGATGAGGCAGAATATTACTGTGTTCTATGG
TACAGCAACCGCTGGGTGTTCGGTGGAGGAACCAAACTGACTGTCCTA 955. PM07A12-H
artificial nt
CAGGTGCAGCTGGTCGAGTCTGGCGGCGGACTGGTCAAGCCTGGCGGCTCCCTG codon
optimized AGACTGTCTTGCGCTGCCTCCGGCTTCACCTTCTCCGACTACTACATGTCCTGGGT
CCGCCAGGCTCCTGGCAAGGGACTGGAATGGGTGGCCTCCATCTCCGACGGCGG
CTCCAACACCTACTACTCCGACATCATCAAGGGCCGGTTCACCATCTCCAGGGACA
ACGCTAAGAACAACCTGTACCTGCAGATGAACTCCCTGAGGGCCGAGGACACCGC
CGTGTACTACTGCGCCAGGGGCTTCCCACTGCTGAGACACGGCGCCTTCGATTACT
GGGGCCAGGGCACCCTGGTCACAGTGTCCTCT 956. PM07A12-L artificial nt
GACATCCAGATGACCCAGTCCCCATCCTCCCTGTCTGCCTCCGTGGGCGACAGAGT codon
optimized GACCATCACATGCCGGGCCTCCCAGAACGTGGACACCAACGTGGCATGGTATCAG
CAGAAGCCAGGCCAGGCCCCTAAGTCCCTGATCTACTCTGCCACCTACCGGTACTC
CGACGTGCCCTCCAGGTTCTCTGGCTCCGCCTCTGGCACCGACTTCACCCTGACCA
TCTCTTCCGTGCAGTCCGAGGACTTCGCTACCTACTACTGCCAGCAGTACAACTCC
TACCCTTACACCTTCGGCGGAGGCACCAAGCTGGAAATCAAG 957. PM07A12-HL
artificial nt
CAGGTGCAGCTGGTCGAGTCTGGCGGCGGACTGGTCAAGCCTGGCGGCTCCCTG codon
optimized AGACTGTCTTGCGCTGCCTCCGGCTTCACCTTCTCCGACTACTACATGTCCTGGGT
CCGCCAGGCTCCTGGCAAGGGACTGGAATGGGTGGCCTCCATCTCCGACGGCGG
CTCCAACACCTACTACTCCGACATCATCAAGGGCCGGTTCACCATCTCCAGGGACA
ACGCTAAGAACAACCTGTACCTGCAGATGAACTCCCTGAGGGCCGAGGACACCGC
CGTGTACTACTGCGCCAGGGGCTTCCCACTGCTGAGACACGGCGCCTTCGATTACT
GGGGCCAGGGCACCCTGGTCACAGTGTCCTCTGGCGGAGGCGGAAGTGGAGGCG
GAGGAAGCGGAGGCGGCGGATCCGACATCCAGATGACCCAGTCCCCATCCTCCCT
GTCTGCCTCCGTGGGCGACAGAGTGACCATCACATGCCGGGCCTCCCAGAACGTG
GACACCAACGTGGCATGGTATCAGCAGAAGCCAGGCCAGGCCCCTAAGTCCCTGA
TCTACTCTGCCACCTACCGGTACTCCGACGTGCCCTCCAGGTTCTCTGGCTCCGCC
TCTGGCACCGACTTCACCCTGACCATCTCTTCCGTGCAGTCCGAGGACTTCGCTAC
CTACTACTGCCAGCAGTACAACTCCTACCCTTACACCTTCGGCGGAGGCACCAAGC
TGGAAATCAAG 958. PM07A12 HL .times. I2C artificial nt
CAGGTGCAGCTGGTCGAGTCTGGCGGCGGACTGGTCAAGCCTGGCGGCTCCCTG HL codon
optimized AGACTGTCTTGCGCTGCCTCCGGCTTCACCTTCTCCGACTACTACATGTCCTGGGT
CCGCCAGGCTCCTGGCAAGGGACTGGAATGGGTGGCCTCCATCTCCGACGGCGG
CTCCAACACCTACTACTCCGACATCATCAAGGGCCGGTTCACCATCTCCAGGGACA
ACGCTAAGAACAACCTGTACCTGCAGATGAACTCCCTGAGGGCCGAGGACACCGC
CGTGTACTACTGCGCCAGGGGCTTCCCACTGCTGAGACACGGCGCCTTCGATTACT
GGGGCCAGGGCACCCTGGTCACAGTGTCCTCTGGCGGAGGCGGAAGTGGAGGCG
GAGGAAGCGGAGGCGGCGGATCCGACATCCAGATGACCCAGTCCCCATCCTCCCT
GTCTGCCTCCGTGGGCGACAGAGTGACCATCACATGCCGGGCCTCCCAGAACGTG
GACACCAACGTGGCATGGTATCAGCAGAAGCCAGGCCAGGCCCCTAAGTCCCTGA
TCTACTCTGCCACCTACCGGTACTCCGACGTGCCCTCCAGGTTCTCTGGCTCCGCC
TCTGGCACCGACTTCACCCTGACCATCTCTTCCGTGCAGTCCGAGGACTTCGCTAC
CTACTACTGCCAGCAGTACAACTCCTACCCTTACACCTTCGGCGGAGGCACCAAGC
TGGAAATCAAGTCCGGCGGAGGGGGCTCTGAAGTGCAGCTGGTGGAAAGCGGAG
GGGGACTGGTGCAGCCCGGGGGAAGTCTGAAGCTGTCCTGTGCCGCCAGCGGCT
TTACCTTCAACAAGTACGCCATGAATTGGGTCCGACAGGCCCCAGGGAAAGGCCTG
GAATGGGTGGCACGGATCCGGTCCAAGTACAACAACTACGCCACCTACTACGCTGA
CTCCGTGAAGGACAGATTCACCATCAGCCGGGACGACTCTAAGAACACCGCCTATC
TGCAGATGAACAACCTGAAAACCGAGGATACAGCTGTGTACTATTGTGTGCGGCAC
GGCAACTTCGGCAACTCCTACATCTCCTACTGGGCCTATTGGGGACAGGGAACACT
GGTCACCGTGTCTAGCGGAGGTGGCGGAAGTGGGGGAGGCGGATCTGGCGGTGG
CGGATCCCAGACCGTGGTCACCCAGGAACCTTCCCTGACCGTCTCCCCAGGCGGC
ACCGTGACCCTGACCTGTGGCTCCTCTACCGGCGCTGTGACCTCCGGCAACTACC
CTAACTGGGTGCAGCAGAAACCCGGACAGGCTCCTAGAGGCCTGATCGGCGGCAC
CAAGTTTCTGGCCCCTGGCACCCCTGCCAGATTCTCCGGCTCCCTGCTGGGAGGC
AAGGCCGCTCTGACCCTGTCTGGCGTGCAGCCTGAGGACGAGGCCGAGTACTACT
GTGTGCTGTGGTACTCCAACAGATGGGTGTTCGGAGGCGGCACAAAGCTGACCGT GCTG 959.
PM07F8-H artificial aa
QVQLVESGGGLVKPGESLRLSCAASGFTFSDYYMSWVRQAPGKGLEWVASISDGGSN
TYYSDIIKGRFTISRDNAKNSLYLQMNSLKAEDTAVYYCARGFPLLRHGAFDYWGQGTL VTVSS
960. PM07F8-HCDR1 artificial aa DYYMS 961. PM07F8-HCDR2 artificial
aa SISDGGSNTYYSDIIKG 962. PM07F8-HCDR3 artificial aa GFPLLRHGAFDY
963. PM07F8-H artificial nt
CAGGTGCAGCTGGTCGAGTCTGGCGGCGGACTGGTGAAGCCTGGCGAGTCCCTG
AGGCTGTCCTGTGCCGCCTCCGGCTTCACCTTCTCCGACTACTACATGAGCTGGGT
CCGCCAGGCCCCTGGGAAGGGGCTGGAATGGGTGGCCTCCATCTCCGACGGCGG
CTCCAACACCTACTACTCCGACATCATCAAGGGCCGGTTCACCATCTCCCGGGACA
ACGCCAAGAACAGCCTGTACCTGCAGATGAACTCCCTGAAGGCCGAGGACACCGC
CGTGTACTACTGCGCCCGGGGCTTCCCTCTGCTGAGACACGGCGCCTTCGATTACT
GGGGCCAGGGCACCCTGGTCACCGTCTCCTCA 964. PM07F8-L artificial aa
DIQMTQSPSSLSASVGDRVTITCRASQNVDTNVAWYQQKPGQAPKSLIYSATYRYSDV
PSRFSGSASGTDFTLTISSVQSEDFATYYCQQYNSYPYTFGGGTKLEIK 965. PM07F8-LCDR1
artificial aa RASQNVDTNVA 966. PM07F8-LCDR2 artificial aa SATYRYS
967. PM07F8-LCDR3 artificial aa QQYNSYPYT 968. PM07F8-L artificial
nt GACATCCAGATGACCCAGTCCCCCAGCTCCCTGTCCGCCTCCGTGGGCGACAGAG
TGACCATCACCTGCAGGGCCTCCCAGAACGTGGACACCAACGTGGCCTGGTATCA
GCAGAAGCCCGGCCAGGCCCCTAAGTCCCTGATCTACTCCGCCACCTACCGGTAC
TCTGACGTGCCTTCCCGGTTCTCCGGCTCCGCGTCCGGCACCGACTTCACCCTGA
CCATCTCCAGCGTGCAGTCTGAGGACTTCGCCACGTACTACTGCCAGCAGTACAAC
TCCTACCCTTACACCTTCGGCGGAGGGACCAAGCTGGAAATCAAG 969. PM07F8-HL
artificial aa
QVQLVESGGGLVKPGESLRLSCAASGFTFSDYYMSWVRQAPGKGLEWVASISDGGSN
TYYSDIIKGRFTISRDNAKNSLYLQMNSLKAEDTAVYYCARGFPLLRHGAFDYWGQGTL
VTVSSGGGGSGGGGSGGGGSDIQMTQSPSSLSASVGDRVTITCRASQNVDTNVAWY
QQKPGQAPKSLIYSATYRYSDVPSRFSGSASGTDFTLTISSVQSEDFATYYCQQYNSYP
YTFGGGTKLEIK 970. PM07F8-HL artificial nt
CAGGTGCAGCTGGTCGAGTCTGGCGGCGGACTGGTGAAGCCTGGCGAGTCCCTG
AGGCTGTCCTGTGCCGCCTCCGGCTTCACCTTCTCCGACTACTACATGAGCTGGGT
CCGCCAGGCCCCTGGGAAGGGGCTGGAATGGGTGGCCTCCATCTCCGACGGCGG
CTCCAACACCTACTACTCCGACATCATCAAGGGCCGGTTCACCATCTCCCGGGACA
ACGCCAAGAACAGCCTGTACCTGCAGATGAACTCCCTGAAGGCCGAGGACACCGC
CGTGTACTACTGCGCCCGGGGCTTCCCTCTGCTGAGACACGGCGCCTTCGATTACT
GGGGCCAGGGCACCCTGGTCACCGTCTCCTCAGGTGGTGGTGGTTCTGGCGGCG
GCGGCTCCGGTGGTGGTGGTTCTGACATCCAGATGACCCAGTCCCCCAGCTCCCT
GTCCGCCTCCGTGGGCGACAGAGTGACCATCACCTGCAGGGCCTCCCAGAACGTG
GACACCAACGTGGCCTGGTATCAGCAGAAGCCCGGCCAGGCCCCTAAGTCCCTGA
TCTACTCCGCCACCTACCGGTACTCTGACGTGCCTTCCCGGTTCTCCGGCTCCGCG
TCCGGCACCGACTTCACCCTGACCATCTCCAGCGTGCAGTCTGAGGACTTCGCCAC
GTACTACTGCCAGCAGTACAACTCCTACCCTTACACCTTCGGCGGAGGGACCAAGC
TGGAAATCAAG 971. PM07F8 HL .times. I2C artificial aa
QVQLVESGGGLVKPGESLRLSCAASGFTFSDYYMSWVRQAPGKGLEWVASISDGGSN HL
TYYSDIIKGRFTISRDNAKNSLYLQMNSLKAEDTAVYYCARGFPLLRHGAFDYWGQGTL
VTVSSGGGGSGGGGSGGGGSDIQMTQSPSSLSASVGDRVTITCRASQNVDTNVAWY
QQKPGQAPKSLIYSATYRYSDVPSRFSGSASGTDFTLTISSVQSEDFATYYCQQYNSYP
YTFGGGTKLEIKSGGGGSEVQLVESGGGLVQPGGSLKLSCAASGFTFNKYAMNWVRQ
APGKGLEWVARIRSKYNNYATYYADSVKDRFTISRDDSKNTAYLQMNNLKTEDTAVYY
CVRHGNFGNSYISYWAYWGQGTLVTVSSGGGGSGGGGSGGGGSQTVVTQEPSLTVS
PGGTVTLTCGSSTGAVTSGNYPNWVQQKPGQAPRGLIGGTKFLAPGTPARFSGSLLG
GKAALTLSGVQPEDEAEYYCVLWYSNRWVFGGGTKLTVL 972. PM07F8 HL .times. I2C
artificial nt
CAGGTGCAGCTGGTCGAGTCTGGCGGCGGACTGGTGAAGCCTGGCGAGTCCCTG HL
AGGCTGTCCTGTGCCGCCTCCGGCTTCACCTTCTCCGACTACTACATGAGCTGGGT
CCGCCAGGCCCCTGGGAAGGGGCTGGAATGGGTGGCCTCCATCTCCGACGGCGG
CTCCAACACCTACTACTCCGACATCATCAAGGGCCGGTTCACCATCTCCCGGGACA
ACGCCAAGAACAGCCTGTACCTGCAGATGAACTCCCTGAAGGCCGAGGACACCGC
CGTGTACTACTGCGCCCGGGGCTTCCCTCTGCTGAGACACGGCGCCTTCGATTACT
GGGGCCAGGGCACCCTGGTCACCGTCTCCTCAGGTGGTGGTGGTTCTGGCGGCG
GCGGCTCCGGTGGTGGTGGTTCTGACATCCAGATGACCCAGTCCCCCAGCTCCCT
GTCCGCCTCCGTGGGCGACAGAGTGACCATCACCTGCAGGGCCTCCCAGAACGTG
GACACCAACGTGGCCTGGTATCAGCAGAAGCCCGGCCAGGCCCCTAAGTCCCTGA
TCTACTCCGCCACCTACCGGTACTCTGACGTGCCTTCCCGGTTCTCCGGCTCCGCG
TCCGGCACCGACTTCACCCTGACCATCTCCAGCGTGCAGTCTGAGGACTTCGCCAC
GTACTACTGCCAGCAGTACAACTCCTACCCTTACACCTTCGGCGGAGGGACCAAGC
TGGAAATCAAGTCCGGAGGTGGTGGATCCGAGGTGCAGCTGGTCGAGTCTGGAGG
AGGATTGGTGCAGCCTGGAGGGTCATTGAAACTCTCATGTGCAGCCTCTGGATTCA
CCTTCAATAAGTACGCCATGAACTGGGTCCGCCAGGCTCCAGGAAAGGGTTTGGAA
TGGGTTGCTCGCATAAGAAGTAAATATAATAATTATGCAACATATTATGCCGATTCAG
TGAAAGACAGGTTCACCATCTCCAGAGATGATTCAAAAAACACTGCCTATCTACAAA
TGAACAACTTGAAAACTGAGGACACTGCCGTGTACTACTGTGTGAGACATGGGAAC
TTCGGTAATAGCTACATATCCTACTGGGCTTACTGGGGCCAAGGGACTCTGGTCAC
CGTCTCCTCAGGTGGTGGTGGTTCTGGCGGCGGCGGCTCCGGTGGTGGTGGTTCT
CAGACTGTTGTGACTCAGGAACCTTCACTCACCGTATCACCTGGTGGAACAGTCAC
ACTCACTTGTGGCTCCTCGACTGGGGCTGTTACATCTGGCAACTACCCAAACTGGG
TCCAACAAAAACCAGGTCAGGCACCCCGTGGTCTAATAGGTGGGACTAAGTTCCTC
GCCCCCGGTACTCCTGCCAGATTCTCAGGCTCCCTGCTTGGAGGCAAGGCTGCCC
TCACCCTCTCAGGGGTACAGCCAGAGGATGAGGCAGAATATTACTGTGTTCTATGG
TACAGCAACCGCTGGGTGTTCGGTGGAGGAACCAAACTGACTGTCCTA 973. PM07F8-H
artificial nt
CAGGTGCAGCTGGTCGAGTCTGGCGGCGGACTGGTCAAGCCTGGCGAGTCCCTGA codon
optimized GACTGTCTTGCGCTGCCTCCGGCTTCACCTTCTCCGACTACTACATGTCCTGGGTC
CGCCAGGCTCCTGGCAAGGGACTGGAATGGGTGGCCTCCATCTCCGACGGCGGCT
CCAACACCTACTACTCCGACATCATCAAGGGCCGGTTCACCATCTCCAGGGACAAC
GCCAAGAACTCCCTGTACCTGCAGATGAACTCCCTGAAGGCCGAGGACACCGCCG
TGTACTACTGCGCCAGGGGCTTCCCACTGCTGAGACACGGCGCCTTCGATTACTGG
GGCCAGGGCACCCTGGTCACAGTGTCCTCT 974. PM07F8-H artificial nt
GACATCCAGATGACCCAGTCCCCATCCTCCCTGTCTGCCTCCGTGGGCGACAGAGT codon
optimized GACCATCACATGCCGGGCCTCCCAGAACGTGGACACCAACGTGGCATGGTATCAG
CAGAAGCCAGGCCAGGCCCCTAAGTCCCTGATCTACTCTGCCACCTACCGGTACTC
CGACGTGCCCTCCAGGTTCTCTGGCTCCGCCTCTGGCACCGACTTCACCCTGACCA
TCTCTTCCGTGCAGTCCGAGGACTTCGCTACCTACTACTGCCAGCAGTACAACTCC
TACCCTTACACCTTCGGCGGAGGCACCAAGCTGGAAATCAAG 975. PM07F8-H artificial
nt CAGGTGCAGCTGGTCGAGTCTGGCGGCGGACTGGTCAAGCCTGGCGAGTCCCTGA codon
optimized GACTGTCTTGCGCTGCCTCCGGCTTCACCTTCTCCGACTACTACATGTCCTGGGTC
CGCCAGGCTCCTGGCAAGGGACTGGAATGGGTGGCCTCCATCTCCGACGGCGGCT
CCAACACCTACTACTCCGACATCATCAAGGGCCGGTTCACCATCTCCAGGGACAAC
GCCAAGAACTCCCTGTACCTGCAGATGAACTCCCTGAAGGCCGAGGACACCGCCG
TGTACTACTGCGCCAGGGGCTTCCCACTGCTGAGACACGGCGCCTTCGATTACTGG
GGCCAGGGCACCCTGGTCACAGTGTCCTCTGGCGGAGGCGGAAGTGGAGGCGGA
GGAAGCGGAGGCGGCGGATCCGACATCCAGATGACCCAGTCCCCATCCTCCCTGT
CTGCCTCCGTGGGCGACAGAGTGACCATCACATGCCGGGCCTCCCAGAACGTGGA
CACCAACGTGGCATGGTATCAGCAGAAGCCAGGCCAGGCCCCTAAGTCCCTGATC
TACTCTGCCACCTACCGGTACTCCGACGTGCCCTCCAGGTTCTCTGGCTCCGCCTC
TGGCACCGACTTCACCCTGACCATCTCTTCCGTGCAGTCCGAGGACTTCGCTACCT
ACTACTGCCAGCAGTACAACTCCTACCCTTACACCTTCGGCGGAGGCACCAAGCTG GAAATCAAG
976. PM07F8 HL .times. I2C artificial nt
CAGGTGCAGCTGGTCGAGTCTGGCGGCGGACTGGTCAAGCCTGGCGAGTCCCTGA HL codon
optimized GACTGTCTTGCGCTGCCTCCGGCTTCACCTTCTCCGACTACTACATGTCCTGGGTC
CGCCAGGCTCCTGGCAAGGGACTGGAATGGGTGGCCTCCATCTCCGACGGCGGCT
CCAACACCTACTACTCCGACATCATCAAGGGCCGGTTCACCATCTCCAGGGACAAC
GCCAAGAACTCCCTGTACCTGCAGATGAACTCCCTGAAGGCCGAGGACACCGCCG
TGTACTACTGCGCCAGGGGCTTCCCACTGCTGAGACACGGCGCCTTCGATTACTGG
GGCCAGGGCACCCTGGTCACAGTGTCCTCTGGCGGAGGCGGAAGTGGAGGCGGA
GGAAGCGGAGGCGGCGGATCCGACATCCAGATGACCCAGTCCCCATCCTCCCTGT
CTGCCTCCGTGGGCGACAGAGTGACCATCACATGCCGGGCCTCCCAGAACGTGGA
CACCAACGTGGCATGGTATCAGCAGAAGCCAGGCCAGGCCCCTAAGTCCCTGATC
TACTCTGCCACCTACCGGTACTCCGACGTGCCCTCCAGGTTCTCTGGCTCCGCCTC
TGGCACCGACTTCACCCTGACCATCTCTTCCGTGCAGTCCGAGGACTTCGCTACCT
ACTACTGCCAGCAGTACAACTCCTACCCTTACACCTTCGGCGGAGGCACCAAGCTG
GAAATCAAGTCCGGCGGAGGGGGCTCTGAAGTGCAGCTGGTGGAAAGCGGAGGG
GGACTGGTGCAGCCCGGGGGAAGTCTGAAGCTGTCCTGTGCCGCCAGCGGCTTTA
CCTTCAACAAGTACGCCATGAATTGGGTCCGACAGGCCCCAGGGAAAGGCCTGGA
ATGGGTGGCACGGATCCGGTCCAAGTACAACAACTACGCCACCTACTACGCTGACT
CCGTGAAGGACAGATTCACCATCAGCCGGGACGACTCTAAGAACACCGCCTATCTG
CAGATGAACAACCTGAAAACCGAGGATACAGCTGTGTACTATTGTGTGCGGCACGG
CAACTTCGGCAACTCCTACATCTCCTACTGGGCCTATTGGGGACAGGGAACACTGG
TCACCGTGTCTAGCGGAGGTGGCGGAAGTGGGGGAGGCGGATCTGGCGGTGGCG
GATCCCAGACCGTGGTCACCCAGGAACCTTCCCTGACCGTCTCCCCAGGCGGCAC
CGTGACCCTGACCTGTGGCTCCTCTACCGGCGCTGTGACCTCCGGCAACTACCCTA
ACTGGGTGCAGCAGAAACCCGGACAGGCTCCTAGAGGCCTGATCGGCGGCACCAA
GTTTCTGGCCCCTGGCACCCCTGCCAGATTCTCCGGCTCCCTGCTGGGAGGCAAG
GCCGCTCTGACCCTGTCTGGCGTGCAGCCTGAGGACGAGGCCGAGTACTACTGTG
TGCTGTGGTACTCCAACAGATGGGTGTTCGGAGGCGGCACAAAGCTGACCGTGCTG 977.
PM07E5-H artificial aa
QVQLVESGGGLVKPGGSLRLSCAASGFTFSDYYMSWVRQAPGKGLEWVASISDGGSN
TYYSDIIKGRFTISRDNAKNNLYLQMNSLRAEDTAVYYCARGFPLLRHGAIDYWGQGTL VTVSS
978. PM07E5-HCDR1 artificial aa DYYMS 979. PM07E5-HCDR2 artificial
aa SISDGGSNTYYSDIIKG 980. PM07E5-HCDR3 artificial aa GFPLLRHGAIDY
981. PM07E5-H artificial nt
CAGGTGCAGCTGGTCGAGTCTGGCGGCGGACTGGTGAAGCCTGGCGGGTCCCTG
AGGCTGTCCTGTGCCGCCTCCGGCTTCACCTTCTCCGACTACTACATGAGCTGGGT
CCGCCAGGCCCCTGGGAAGGGGCTGGAATGGGTGGCCTCCATCTCCGACGGCGG
CTCCAACACCTACTACTCCGACATCATCAAGGGCCGGTTCACCATCTCCCGAGACA
ACGCCAAGAACAATCTGTACCTGCAGATGAACTCCCTGAGGGCCGAGGACACCGC
CGTGTACTACTGCGCCCGGGGCTTCCCTCTGCTGAGACACGGCGCCATCGATTACT
GGGGCCAGGGCACCCTGGTCACCGTCTCCTCA 982. PM07E5-L artificial aa
DIQMTQSPSSLSASVGDRVTITCRASQNVDTNVAWYQQKPGQAPKSLIYSATYRYSDV
PSRFSGSASGTDFTLTISSVQSEDFATYYCQQYNSYPYTFGGGTKLEIK 983. PM07E5-LCDR1
artificial aa RASQNVDTNVA 984. PM07E5-LCDR2 artificial aa SATYRYS
985. PM07E5-LCDR3 artificial aa QQYNSYPYT 986. PM07E5-L artificial
nt GACATCCAGATGACCCAGTCCCCCAGCTCCCTGTCCGCCTCCGTGGGCGACAGAG
TGACCATCACCTGCAGGGCCTCCCAGAACGTGGACACCAACGTGGCCTGGTATCA
GCAGAAGCCCGGCCAGGCCCCTAAGTCCCTGATCTACTCCGCCACCTACCGGTAC
TCTGACGTGCCTTCCCGGTTCTCCGGCTCCGCGTCCGGCACCGACTTCACCCTGA
CCATCTCCAGCGTGCAGTCTGAGGACTTCGCCACGTACTACTGCCAGCAGTACAAC
TCCTACCCTTACACCTTCGGCGGAGGGACCAAGCTGGAAATCAAG 987. PM07E5-HL
artificial aa
QVQLVESGGGLVKPGGSLRLSCAASGFTFSDYYMSWVRQAPGKGLEWVASISDGGSN
TYYSDIIKGRFTISRDNAKNNLYLQMNSLRAEDTAVYYCARGFPLLRHGAIDYWGQGTL
VTVSSGGGGSGGGGSGGGGSDIQMTQSPSSLSASVGDRVTITCRASQNVDTNVAWY
QQKPGQAPKSLIYSATYRYSDVPSRFSGSASGTDFTLTISSVQSEDFATYYCQQYNSYP
YTFGGGTKLEIK 988. PM07E5-HL artificial nt
CAGGTGCAGCTGGTCGAGTCTGGCGGCGGACTGGTGAAGCCTGGCGGGTCCCTG
AGGCTGTCCTGTGCCGCCTCCGGCTTCACCTTCTCCGACTACTACATGAGCTGGGT
CCGCCAGGCCCCTGGGAAGGGGCTGGAATGGGTGGCCTCCATCTCCGACGGCGG
CTCCAACACCTACTACTCCGACATCATCAAGGGCCGGTTCACCATCTCCCGAGACA
ACGCCAAGAACAATCTGTACCTGCAGATGAACTCCCTGAGGGCCGAGGACACCGC
CGTGTACTACTGCGCCCGGGGCTTCCCTCTGCTGAGACACGGCGCCATCGATTACT
GGGGCCAGGGCACCCTGGTCACCGTCTCCTCAGGTGGTGGTGGTTCTGGCGGCG
GCGGCTCCGGTGGTGGTGGTTCTGACATCCAGATGACCCAGTCCCCCAGCTCCCT
GTCCGCCTCCGTGGGCGACAGAGTGACCATCACCTGCAGGGCCTCCCAGAACGTG
GACACCAACGTGGCCTGGTATCAGCAGAAGCCCGGCCAGGCCCCTAAGTCCCTGA
TCTACTCCGCCACCTACCGGTACTCTGACGTGCCTTCCCGGTTCTCCGGCTCCGCG
TCCGGCACCGACTTCACCCTGACCATCTCCAGCGTGCAGTCTGAGGACTTCGCCAC
GTACTACTGCCAGCAGTACAACTCCTACCCTTACACCTTCGGCGGAGGGACCAAGC
TGGAAATCAAG 989. PM07E5 HL .times. I2C artificial aa
QVQLVESGGGLVKPGGSLRLSCAASGFTFSDYYMSWVRQAPGKGLEWVASISDGGSN HL
TYYSDIIKGRFTISRDNAKNNLYLQMNSLRAEDTAVYYCARGFPLLRHGAIDYWGQGTL
VTVSSGGGGSGGGGSGGGGSDIQMTQSPSSLSASVGDRVTITCRASQNVDTNVAWY
QQKPGQAPKSLIYSATYRYSDVPSRFSGSASGTDFTLTISSVQSEDFATYYCQQYNSYP
YTFGGGTKLEIKSGGGGSEVQLVESGGGLVQPGGSLKLSCAASGFTFNKYAMNWVRQ
APGKGLEWVARIRSKYNNYATYYADSVKDRFTISRDDSKNTAYLQMNNLKTEDTAVYY
CVRHGNFGNSYISYWAYWGQGTLVTVSSGGGGSGGGGSGGGGSQTVVTQEPSLTVS
PGGTVTLTCGSSTGAVTSGNYPNWVQQKPGQAPRGLIGGTKFLAPGTPARFSGSLLG
GKAALTLSGVQPEDEAEYYCVLWYSNRWVFGGGTKLTVL 990. PM07E5 HL .times. I2C
artificial nt
CAGGTGCAGCTGGTCGAGTCTGGCGGCGGACTGGTGAAGCCTGGCGGGTCCCTG HL
AGGCTGTCCTGTGCCGCCTCCGGCTTCACCTTCTCCGACTACTACATGAGCTGGGT
CCGCCAGGCCCCTGGGAAGGGGCTGGAATGGGTGGCCTCCATCTCCGACGGCGG
CTCCAACACCTACTACTCCGACATCATCAAGGGCCGGTTCACCATCTCCCGAGACA
ACGCCAAGAACAATCTGTACCTGCAGATGAACTCCCTGAGGGCCGAGGACACCGC
CGTGTACTACTGCGCCCGGGGCTTCCCTCTGCTGAGACACGGCGCCATCGATTACT
GGGGCCAGGGCACCCTGGTCACCGTCTCCTCAGGTGGTGGTGGTTCTGGCGGCG
GCGGCTCCGGTGGTGGTGGTTCTGACATCCAGATGACCCAGTCCCCCAGCTCCCT
GTCCGCCTCCGTGGGCGACAGAGTGACCATCACCTGCAGGGCCTCCCAGAACGTG
GACACCAACGTGGCCTGGTATCAGCAGAAGCCCGGCCAGGCCCCTAAGTCCCTGA
TCTACTCCGCCACCTACCGGTACTCTGACGTGCCTTCCCGGTTCTCCGGCTCCGCG
TCCGGCACCGACTTCACCCTGACCATCTCCAGCGTGCAGTCTGAGGACTTCGCCAC
GTACTACTGCCAGCAGTACAACTCCTACCCTTACACCTTCGGCGGAGGGACCAAGC
TGGAAATCAAGTCCGGAGGTGGTGGATCCGAGGTGCAGCTGGTCGAGTCTGGAGG
AGGATTGGTGCAGCCTGGAGGGTCATTGAAACTCTCATGTGCAGCCTCTGGATTCA
CCTTCAATAAGTACGCCATGAACTGGGTCCGCCAGGCTCCAGGAAAGGGTTTGGAA
TGGGTTGCTCGCATAAGAAGTAAATATAATAATTATGCAACATATTATGCCGATTCAG
TGAAAGACAGGTTCACCATCTCCAGAGATGATTCAAAAAACACTGCCTATCTACAAA
TGAACAACTTGAAAACTGAGGACACTGCCGTGTACTACTGTGTGAGACATGGGAAC
TTCGGTAATAGCTACATATCCTACTGGGCTTACTGGGGCCAAGGGACTCTGGTCAC
CGTCTCCTCAGGTGGTGGTGGTTCTGGCGGCGGCGGCTCCGGTGGTGGTGGTTCT
CAGACTGTTGTGACTCAGGAACCTTCACTCACCGTATCACCTGGTGGAACAGTCAC
ACTCACTTGTGGCTCCTCGACTGGGGCTGTTACATCTGGCAACTACCCAAACTGGG
TCCAACAAAAACCAGGTCAGGCACCCCGTGGTCTAATAGGTGGGACTAAGTTCCTC
GCCCCCGGTACTCCTGCCAGATTCTCAGGCTCCCTGCTTGGAGGCAAGGCTGCCC
TCACCCTCTCAGGGGTACAGCCAGAGGATGAGGCAGAATATTACTGTGTTCTATGG
TACAGCAACCGCTGGGTGTTCGGTGGAGGAACCAAACTGACTGTCCTA 991. PM07D3-H
artificial aa
QVQLVESGGGLVKPGESLRLSCAASGFTFSDYYMSWVRQAPGKGLEWVASISDGGSN
TYYSDIIKGRFTISRDNAKNSLYLQMNSLKAEDTAVYYCARGFPLLRHGAIDYWGQGTLV TVSS
992. PM07D3-HCDR1 artificial aa DYYMS 993. PM07D3-HCDR2 artificial
aa SISDGGSNTYYSDIIKG 994. PM07D3-HCDR3 artificial aa GFPLLRHGAIDY
995. PM07D3-H artificial nt
CAGGTGCAGCTGGTCGAGTCTGGCGGCGGACTGGTGAAGCCTGGCGAGTCCCTG
AGGCTGTCCTGTGCCGCCTCCGGCTTCACCTTCTCCGACTACTACATGAGCTGGGT
CCGCCAGGCCCCTGGGAAGGGGCTGGAATGGGTGGCCTCCATCTCCGACGGCGG
CTCCAACACCTACTACTCCGACATCATCAAGGGCCGGTTCACCATCTCCCGAGACA
ACGCCAAGAACAGCCTGTACCTGCAGATGAACTCCCTGAAGGCCGAGGACACCGC
CGTGTACTACTGCGCCCGGGGCTTCCCTCTGCTGAGACACGGCGCCATCGATTACT
GGGGCCAGGGCACCCTGGTCACCGTCTCCTCA 996. PM07D3-L artificial aa
DIQMTQSPSSLSASVGDRVTITCRASQNVDTNVAWYQQKPGQAPKSLIYSATYRYSDV
PSRFSGSASGTDFTLTISSVQSEDFATYYCQQYNSYPYTFGGGTKLEIK 997. PM07D3-LCDR1
artificial aa RASQNVDTNVA 998. PM07D3-LCDR2 artificial aa SATYRYS
999. PM07D3-LCDR3 artificial aa QQYNSYPYT 1000. PM07D3-L artificial
nt GACATCCAGATGACCCAGTCCCCCAGCTCCCTGTCCGCCTCCGTGGGCGACAGAG
TGACCATCACCTGCAGGGCCTCCCAGAACGTGGACACCAACGTGGCCTGGTATCA
GCAGAAGCCCGGCCAGGCCCCTAAGTCCCTGATCTACTCCGCCACCTACCGGTAC
TCTGACGTGCCTTCCCGGTTCTCCGGCTCCGCGTCCGGCACCGACTTCACCCTGA
CCATCTCCAGCGTGCAGTCTGAGGACTTCGCCACGTACTACTGCCAGCAGTACAAC
TCCTACCCTTACACCTTCGGCGGAGGGACCAAGCTGGAAATCAAG 1001. PM07D3-HL
artificial aa
QVQLVESGGGLVKPGESLRLSCAASGFTFSDYYMSWVRQAPGKGLEWVASISDGGSN
TYYSDIIKGRFTISRDNAKNSLYLQMNSLKAEDTAVYYCARGFPLLRHGAIDYWGQGTLV
TVSSGGGGSGGGGSGGGGSDIQMTQSPSSLSASVGDRVTITCRASQNVDTNVAWYQ
QKPGQAPKSLIYSATYRYSDVPSRFSGSASGTDFTLTISSVQSEDFATYYCQQYNSYPY
TFGGGTKLEIK 1002. PM07D3-HL artificial nt
CAGGTGCAGCTGGTCGAGTCTGGCGGCGGACTGGTGAAGCCTGGCGAGTCCCTG
AGGCTGTCCTGTGCCGCCTCCGGCTTCACCTTCTCCGACTACTACATGAGCTGGGT
CCGCCAGGCCCCTGGGAAGGGGCTGGAATGGGTGGCCTCCATCTCCGACGGCGG
CTCCAACACCTACTACTCCGACATCATCAAGGGCCGGTTCACCATCTCCCGAGACA
ACGCCAAGAACAGCCTGTACCTGCAGATGAACTCCCTGAAGGCCGAGGACACCGC
CGTGTACTACTGCGCCCGGGGCTTCCCTCTGCTGAGACACGGCGCCATCGATTACT
GGGGCCAGGGCACCCTGGTCACCGTCTCCTCAGGTGGTGGTGGTTCTGGCGGCG
GCGGCTCCGGTGGTGGTGGTTCTGACATCCAGATGACCCAGTCCCCCAGCTCCCT
GTCCGCCTCCGTGGGCGACAGAGTGACCATCACCTGCAGGGCCTCCCAGAACGTG
GACACCAACGTGGCCTGGTATCAGCAGAAGCCCGGCCAGGCCCCTAAGTCCCTGA
TCTACTCCGCCACCTACCGGTACTCTGACGTGCCTTCCCGGTTCTCCGGCTCCGCG
TCCGGCACCGACTTCACCCTGACCATCTCCAGCGTGCAGTCTGAGGACTTCGCCAC
GTACTACTGCCAGCAGTACAACTCCTACCCTTACACCTTCGGCGGAGGGACCAAGC
TGGAAATCAAG 1003. PM07D3 HL .times. I2C artificial aa
QVQLVESGGGLVKPGESLRLSCAASGFTFSDYYMSWVRQAPGKGLEWVASISDGGSN HL
TYYSDIIKGRFTISRDNAKNSLYLQMNSLKAEDTAVYYCARGFPLLRHGAIDYWGQGTLV
TVSSGGGGSGGGGSGGGGSDIQMTQSPSSLSASVGDRVTITCRASQNVDTNVAWYQ
QKPGQAPKSLIYSATYRYSDVPSRFSGSASGTDFTLTISSVQSEDFATYYCQQYNSYPY
TFGGGTKLEIKSGGGGSEVQLVESGGGLVQPGGSLKLSCAASGFTFNKYAMNWVRQA
PGKGLEWVARIRSKYNNYATYYADSVKDRFTISRDDSKNTAYLQMNNLKTEDTAVYYC
VRHGNFGNSYISYWAYWGQGTLVTVSSGGGGSGGGGSGGGGSQTVVTQEPSLTVSP
GGTVTLTCGSSTGAVTSGNYPNWVQQKPGQAPRGLIGGTKFLAPGTPARFSGSLLGG
KAALTLSGVQPEDEAEYYCVLWYSNRWVFGGGTKLTVL 1004. PM07D3 HL .times. I2C
artificial nt
CAGGTGCAGCTGGTCGAGTCTGGCGGCGGACTGGTGAAGCCTGGCGAGTCCCTG HL
AGGCTGTCCTGTGCCGCCTCCGGCTTCACCTTCTCCGACTACTACATGAGCTGGGT
CCGCCAGGCCCCTGGGAAGGGGCTGGAATGGGTGGCCTCCATCTCCGACGGCGG
CTCCAACACCTACTACTCCGACATCATCAAGGGCCGGTTCACCATCTCCCGAGACA
ACGCCAAGAACAGCCTGTACCTGCAGATGAACTCCCTGAAGGCCGAGGACACCGC
CGTGTACTACTGCGCCCGGGGCTTCCCTCTGCTGAGACACGGCGCCATCGATTACT
GGGGCCAGGGCACCCTGGTCACCGTCTCCTCAGGTGGTGGTGGTTCTGGCGGCG
GCGGCTCCGGTGGTGGTGGTTCTGACATCCAGATGACCCAGTCCCCCAGCTCCCT
GTCCGCCTCCGTGGGCGACAGAGTGACCATCACCTGCAGGGCCTCCCAGAACGTG
GACACCAACGTGGCCTGGTATCAGCAGAAGCCCGGCCAGGCCCCTAAGTCCCTGA
TCTACTCCGCCACCTACCGGTACTCTGACGTGCCTTCCCGGTTCTCCGGCTCCGCG
TCCGGCACCGACTTCACCCTGACCATCTCCAGCGTGCAGTCTGAGGACTTCGCCAC
GTACTACTGCCAGCAGTACAACTCCTACCCTTACACCTTCGGCGGAGGGACCAAGC
TGGAAATCAAGTCCGGAGGTGGTGGATCCGAGGTGCAGCTGGTCGAGTCTGGAGG
AGGATTGGTGCAGCCTGGAGGGTCATTGAAACTCTCATGTGCAGCCTCTGGATTCA
CCTTCAATAAGTACGCCATGAACTGGGTCCGCCAGGCTCCAGGAAAGGGTTTGGAA
TGGGTTGCTCGCATAAGAAGTAAATATAATAATTATGCAACATATTATGCCGATTCAG
TGAAAGACAGGTTCACCATCTCCAGAGATGATTCAAAAAACACTGCCTATCTACAAA
TGAACAACTTGAAAACTGAGGACACTGCCGTGTACTACTGTGTGAGACATGGGAAC
TTCGGTAATAGCTACATATCCTACTGGGCTTACTGGGGCCAAGGGACTCTGGTCAC
CGTCTCCTCAGGTGGTGGTGGTTCTGGCGGCGGCGGCTCCGGTGGTGGTGGTTCT
CAGACTGTTGTGACTCAGGAACCTTCACTCACCGTATCACCTGGTGGAACAGTCAC
ACTCACTTGTGGCTCCTCGACTGGGGCTGTTACATCTGGCAACTACCCAAACTGGG
TCCAACAAAAACCAGGTCAGGCACCCCGTGGTCTAATAGGTGGGACTAAGTTCCTC
GCCCCCGGTACTCCTGCCAGATTCTCAGGCTCCCTGCTTGGAGGCAAGGCTGCCC
TCACCCTCTCAGGGGTACAGCCAGAGGATGAGGCAGAATATTACTGTGTTCTATGG
TACAGCAACCGCTGGGTGTTCGGTGGAGGAACCAAACTGACTGTCCTA 1005. PM26C9-H
artificial aa
QVQLVQSGPEVVKPGASVKVSCKASGYTFTGYVMHWVRQTPGQRLEWIGYINPYNDV
TRYNGKFKGRVTITSDKSSSTAYMELSSLRSEDTAVYYCARGENWYYFDSWGRGTLVT VSS
1006. PM26C9-HCDR1 artificial aa GYVMH 1007. PM26C9-HCDR2
artificial aa YINPYNDVTRYNGKFKG 1008. PM26C9-HCDR3 artificial aa
GENWYYFDS 1009. PM26C9-H artificial nt
CAGGTGCAGCTGGTCCAGTCCGGCCCTGAGGTGGTGAAGCCTGGCGCCTCCGTGA
AGGTGTCCTGCAAGGCCTCCGGCTACACCTTCACCGGCTACGTGATGCACTGGGT
GAGACAGACACCCGGCCAGCGGCTGGAATGGATCGGCTACATCAACCCTTACAAC
GACGTGACCCGGTACAACGGCAAGTTCAAGGGCAGAGTCACCATTACCAGCGACA
AGTCCTCCTCCACCGCCTACATGGAACTGTCCAGCCTGAGGTCTGAGGACACCGC
CGTGTACTACTGCGCCAGGGGCGAGAACTGGTACTACTTCGACTCCTGGGGCAGA
GGCACTCTGGTCACCGTCTCCTCC 1010. PM26C9-L artificial aa
DVVMTQSPLSLAVTLGQPASISCRASESIDSYDNTFMHWYQQRPGQSPSLLIYRASILQ
SGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCHQSIEDPYTFGGGTKLEIK 1011.
PM26C9-LCDR1 artificial aa RASESIDSYDNTFMH 1012. PM26C9-LCDR2
artificial aa RASILQS 1013. PM26C9-LCDR3 artificial aa HQSIEDPYT
1014. PM26C9-L artificial nt
GACGTCGTGATGACCCAGTCTCCACTCTCCCTGGCTGTGACTCTGGGCCAGCCGG
CCTCCATCTCTTGCCGGGCCTCCGAGTCCATCGACTCCTACGACAACACCTTCATG
CACTGGTATCAGCAGAGGCCTGGCCAGTCTCCTAGCCTGCTGATCTACCGGGCCT
CTATCCTGCAATCCGGCGTCCCTGACCGGTTCTCCGGCTCTGGCTCCGGTACCGA
CTTCACCCTGAAAATCTCCCGTGTGGAGGCCGAGGACGTGGGCGTCTACTACTGC
CACCAGTCCATCGAGGACCCTTACACCTTCGGCGGAGGGACCAAGCTGGAAATCA AG
1015. PM26C9-LH artificial aa
DVVMTQSPLSLAVTLGQPASISCRASESIDSYDNTFMHWYQQRPGQSPSLLIYRASILQ
SGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCHQSIEDPYTFGGGTKLEIKGGGGSG
GGGSGGGGSQVQLVQSGPEVVKPGASVKVSCKASGYTFTGYVMHWVRQTPGQRLE
WIGYINPYNDVTRYNGKFKGRVTITSDKSSSTAYMELSSLRSEDTAVYYCARGENWYYF
DSWGRGTLVTVSS 1016. PM26C9-LH artificial nt
GACGTCGTGATGACCCAGTCTCCACTCTCCCTGGCTGTGACTCTGGGCCAGCCGG
CCTCCATCTCTTGCCGGGCCTCCGAGTCCATCGACTCCTACGACAACACCTTCATG
CACTGGTATCAGCAGAGGCCTGGCCAGTCTCCTAGCCTGCTGATCTACCGGGCCT
CTATCCTGCAATCCGGCGTCCCTGACCGGTTCTCCGGCTCTGGCTCCGGTACCGA
CTTCACCCTGAAAATCTCCCGTGTGGAGGCCGAGGACGTGGGCGTCTACTACTGC
CACCAGTCCATCGAGGACCCTTACACCTTCGGCGGAGGGACCAAGCTGGAAATCA
AGGGTGGTGGTGGTTCTGGCGGCGGCGGCTCCGGTGGTGGTGGTTCTCAGGTGC
AGCTGGTCCAGTCCGGCCCTGAGGTGGTGAAGCCTGGCGCCTCCGTGAAGGTGTC
CTGCAAGGCCTCCGGCTACACCTTCACCGGCTACGTGATGCACTGGGTGAGACAG
ACACCCGGCCAGCGGCTGGAATGGATCGGCTACATCAACCCTTACAACGACGTGA
CCCGGTACAACGGCAAGTTCAAGGGCAGAGTCACCATTACCAGCGACAAGTCCTCC
TCCACCGCCTACATGGAACTGTCCAGCCTGAGGTCTGAGGACACCGCCGTGTACTA
CTGCGCCAGGGGCGAGAACTGGTACTACTTCGACTCCTGGGGCAGAGGCACTCTG
GTCACCGTCTCCTCC 1017. PM26C9 LH .times. I2C artificial aa
DVVMTQSPLSLAVTLGQPASISCRASESIDSYDNTFMHWYQQRPGQSPSLLIYRASILQ HL
SGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCHQSIEDPYTFGGGTKLEIKGGGGSG
GGGSGGGGSQVQLVQSGPEVVKPGASVKVSCKASGYTFTGYVMHWVRQTPGQRLE
WIGYINPYNDVTRYNGKFKGRVTITSDKSSSTAYMELSSLRSEDTAVYYCARGENWYYF
DSWGRGTLVTVSSGGGGSEVQLVESGGGLVQPGGSLKLSCAASGFTFNKYAMNWVR
QAPGKGLEWVARIRSKYNNYATYYADSVKDRFTISRDDSKNTAYLQMNNLKTEDTAVY
YCVRHGNFGNSYISYWAYWGQGTLVTVSSGGGGSGGGGSGGGGSQTVVTQEPSLTV
SPGGTVTLTCGSSTGAVTSGNYPNWVQQKPGQAPRGLIGGTKFLAPGTPARFSGSLL
GGKAALTLSGVQPEDEAEYYCVLWYSNRWVFGGGTKLTVL 1018. PM26C9 LH .times.
I2C artificial nt
GACGTCGTGATGACCCAGTCTCCACTCTCCCTGGCTGTGACTCTGGGCCAGCCGG HL
CCTCCATCTCTTGCCGGGCCTCCGAGTCCATCGACTCCTACGACAACACCTTCATG
CACTGGTATCAGCAGAGGCCTGGCCAGTCTCCTAGCCTGCTGATCTACCGGGCCT
CTATCCTGCAATCCGGCGTCCCTGACCGGTTCTCCGGCTCTGGCTCCGGTACCGA
CTTCACCCTGAAAATCTCCCGTGTGGAGGCCGAGGACGTGGGCGTCTACTACTGC
CACCAGTCCATCGAGGACCCTTACACCTTCGGCGGAGGGACCAAGCTGGAAATCA
AGGGTGGTGGTGGTTCTGGCGGCGGCGGCTCCGGTGGTGGTGGTTCTCAGGTGC
AGCTGGTCCAGTCCGGCCCTGAGGTGGTGAAGCCTGGCGCCTCCGTGAAGGTGTC
CTGCAAGGCCTCCGGCTACACCTTCACCGGCTACGTGATGCACTGGGTGAGACAG
ACACCCGGCCAGCGGCTGGAATGGATCGGCTACATCAACCCTTACAACGACGTGA
CCCGGTACAACGGCAAGTTCAAGGGCAGAGTCACCATTACCAGCGACAAGTCCTCC
TCCACCGCCTACATGGAACTGTCCAGCCTGAGGTCTGAGGACACCGCCGTGTACTA
CTGCGCCAGGGGCGAGAACTGGTACTACTTCGACTCCTGGGGCAGAGGCACTCTG
GTCACCGTCTCCTCCGGAGGTGGTGGATCCGAGGTGCAGCTGGTCGAGTCTGGAG
GAGGATTGGTGCAGCCTGGAGGGTCATTGAAACTCTCATGTGCAGCCTCTGGATTC
ACCTTCAATAAGTACGCCATGAACTGGGTCCGCCAGGCTCCAGGAAAGGGTTTGGA
ATGGGTTGCTCGCATAAGAAGTAAATATAATAATTATGCAACATATTATGCCGATTCA
GTGAAAGACAGGTTCACCATCTCCAGAGATGATTCAAAAAACACTGCCTATCTACAA
ATGAACAACTTGAAAACTGAGGACACTGCCGTGTACTACTGTGTGAGACATGGGAA
CTTCGGTAATAGCTACATATCCTACTGGGCTTACTGGGGCCAAGGGACTCTGGTCA
CCGTCTCCTCAGGTGGTGGTGGTTCTGGCGGCGGCGGCTCCGGTGGTGGTGGTTC
TCAGACTGTTGTGACTCAGGAACCTTCACTCACCGTATCACCTGGTGGAACAGTCA
CACTCACTTGTGGCTCCTCGACTGGGGCTGTTACATCTGGCAACTACCCAAACTGG
GTCCAACAAAAACCAGGTCAGGCACCCCGTGGTCTAATAGGTGGGACTAAGTTCCT
CGCCCCCGGTACTCCTGCCAGATTCTCAGGCTCCCTGCTTGGAGGCAAGGCTGCC
CTCACCCTCTCAGGGGTACAGCCAGAGGATGAGGCAGAATATTACTGTGTTCTATG
GTACAGCAACCGCTGGGTGTTCGGTGGAGGAACCAAACTGACTGTCCTA 1019. PM26H4-H
artificial aa
QVQLVQSGPEVVKPGASVKVSCKASGYTFTGYVLHWVKQTPGQRLEWIGYINPYNDVT
RYNGKFKGRVTITSDTSASTAYMELSGLTSEDTAVYYCARGENWYYFDSWGRGTLVTV SS 1020.
PM26H4-HCDR1 artificial aa GYVLH 1021. PM26H4-HCDR2 artificial aa
YINPYNDVTRYNGKFKG 1022. PM26H4-HCDR3 artificial aa GENWYYFDS 1023.
PM26H4-H artificial nt
CAGGTGCAGCTGGTCCAGTCCGGCCCTGAGGTGGTGAAGCCTGGCGCCTCCGTGA
AGGTGTCCTGCAAGGCCTCCGGCTACACCTTCACCGGCTACGTGCTGCACTGGGT
GAAACAGACACCCGGCCAGCGGCTGGAATGGATCGGCTACATCAACCCTTACAAC
GACGTGACCCGGTACAACGGCAAGTTCAAGGGCAGAGTCACCATTACCAGCGACA
CGTCCGCCTCCACCGCCTACATGGAACTGTCCGGCCTGACGTCTGAGGACACCGC
CGTGTATTACTGCGCCAGGGGCGAGAACTGGTACTACTTCGACTCCTGGGGCAGA
GGCACTCTGGTCACCGTCTCCTCC 1024. PM26H4-L artificial aa
DVVMTQSPLSLAVTLGQRASISCRASESIDSYGNTFMHWYQQRPGQSPRLLIYRASILE
SGVPARFSGSGSGTDFTLAISRVEAEDVGVYYCHQSIEDPYTFGGGTKLEIK 1025.
PM26H4-LCDR1 artificial aa RASESIDSYGNTFMH 1026. PM26H4-LCDR2
artificial aa RASILES 1027. PM26H4-LCDR3 artificial aa HQSIEDPYT
1028. PM26H4-L artificial nt
GACGTCGTGATGACCCAGTCTCCACTCTCCCTGGCTGTGACTCTGGGCCAGCGGG
CCTCCATCTCTTGCCGGGCCTCCGAGTCCATCGACTCCTACGGCAACACCTTCATG
CACTGGTATCAGCAGAGGCCTGGCCAGTCTCCTCGCCTGCTGATCTACCGGGCCT
CTATCCTGGAATCCGGCGTCCCTGCCCGGTTCTCCGGCTCTGGCTCCGGCACCGA
CTTCACCCTGGCAATCTCCCGTGTGGAGGCCGAGGACGTGGGCGTCTACTACTGC
CACCAGTCCATCGAGGACCCTTACACCTTCGGCGGAGGGACCAAGCTGGAAATCA AG 1029.
PM26H4-LH artificial aa
DVVMTQSPLSLAVTLGQRASISCRASESIDSYGNTFMHWYQQRPGQSPRLLIYRASILE
SGVPARFSGSGSGTDFTLAISRVEAEDVGVYYCHQSIEDPYTFGGGTKLEIKGGGGSG
GGGSGGGGSQVQLVQSGPEVVKPGASVKVSCKASGYTFTGYVLHWVKQTPGQRLEW
IGYINPYNDVTRYNGKFKGRVTITSDTSASTAYMELSGLTSEDTAVYYCARGENWYYFD
SWGRGTLVTVSS 1030. PM26H4-LH artificial nt
GACGTCGTGATGACCCAGTCTCCACTCTCCCTGGCTGTGACTCTGGGCCAGCGGG
CCTCCATCTCTTGCCGGGCCTCCGAGTCCATCGACTCCTACGGCAACACCTTCATG
CACTGGTATCAGCAGAGGCCTGGCCAGTCTCCTCGCCTGCTGATCTACCGGGCCT
CTATCCTGGAATCCGGCGTCCCTGCCCGGTTCTCCGGCTCTGGCTCCGGCACCGA
CTTCACCCTGGCAATCTCCCGTGTGGAGGCCGAGGACGTGGGCGTCTACTACTGC
CACCAGTCCATCGAGGACCCTTACACCTTCGGCGGAGGGACCAAGCTGGAAATCA
AGGGTGGTGGTGGTTCTGGCGGCGGCGGCTCCGGTGGTGGTGGTTCTCAGGTGC
AGCTGGTCCAGTCCGGCCCTGAGGTGGTGAAGCCTGGCGCCTCCGTGAAGGTGTC
CTGCAAGGCCTCCGGCTACACCTTCACCGGCTACGTGCTGCACTGGGTGAAACAG
ACACCCGGCCAGCGGCTGGAATGGATCGGCTACATCAACCCTTACAACGACGTGA
CCCGGTACAACGGCAAGTTCAAGGGCAGAGTCACCATTACCAGCGACACGTCCGC
CTCCACCGCCTACATGGAACTGTCCGGCCTGACGTCTGAGGACACCGCCGTGTATT
ACTGCGCCAGGGGCGAGAACTGGTACTACTTCGACTCCTGGGGCAGAGGCACTCT
GGTCACCGTCTCCTCC 1031. PM26H4 LH .times. I2C artificial aa
DVVMTQSPLSLAVTLGQRASISCRASESIDSYGNTFMHWYQQRPGQSPRLLIYRASILE HL
SGVPARFSGSGSGTDFTLAISRVEAEDVGVYYCHQSIEDPYTFGGGTKLEIKGGGGSG
GGGSGGGGSQVQLVQSGPEVVKPGASVKVSCKASGYTFTGYVLHWVKQTPGQRLEW
IGYINPYNDVTRYNGKFKGRVTITSDTSASTAYMELSGLTSEDTAVYYCARGENWYYFD
SWGRGTLVTVSSSGGGGSEVQLVESGGGLVQPGGSLKLSCAASGFTFNKYAMNWVR
QAPGKGLEWVARIRSKYNNYATYYADSVKDRFTISRDDSKNTAYLQMNNLKTEDTAVY
YCVRHGNFGNSYISYWAYWGQGTLVTVSSGGGGSGGGGSGGGGSQTVVTQEPSLTV
SPGGTVTLTCGSSTGAVTSGNYPNWVQQKPGQAPRGLIGGTKFLAPGTPARFSGSLL
GGKAALTLSGVQPEDEAEYYCVLWYSNRWVFGGGTKLTVL 1032. PM26H4 LH .times.
I2C artificial nt
GACGTCGTGATGACCCAGTCTCCACTCTCCCTGGCTGTGACTCTGGGCCAGCGGG HL
CCTCCATCTCTTGCCGGGCCTCCGAGTCCATCGACTCCTACGGCAACACCTTCATG
CACTGGTATCAGCAGAGGCCTGGCCAGTCTCCTCGCCTGCTGATCTACCGGGCCT
CTATCCTGGAATCCGGCGTCCCTGCCCGGTTCTCCGGCTCTGGCTCCGGCACCGA
CTTCACCCTGGCAATCTCCCGTGTGGAGGCCGAGGACGTGGGCGTCTACTACTGC
CACCAGTCCATCGAGGACCCTTACACCTTCGGCGGAGGGACCAAGCTGGAAATCA
AGGGTGGTGGTGGTTCTGGCGGCGGCGGCTCCGGTGGTGGTGGTTCTCAGGTGC
AGCTGGTCCAGTCCGGCCCTGAGGTGGTGAAGCCTGGCGCCTCCGTGAAGGTGTC
CTGCAAGGCCTCCGGCTACACCTTCACCGGCTACGTGCTGCACTGGGTGAAACAG
ACACCCGGCCAGCGGCTGGAATGGATCGGCTACATCAACCCTTACAACGACGTGA
CCCGGTACAACGGCAAGTTCAAGGGCAGAGTCACCATTACCAGCGACACGTCCGC
CTCCACCGCCTACATGGAACTGTCCGGCCTGACGTCTGAGGACACCGCCGTGTATT
ACTGCGCCAGGGGCGAGAACTGGTACTACTTCGACTCCTGGGGCAGAGGCACTCT
GGTCACCGTCTCCTCCGGAGGTGGTGGATCCGAGGTGCAGCTGGTCGAGTCTGGA
GGAGGATTGGTGCAGCCTGGAGGGTCATTGAAACTCTCATGTGCAGCCTCTGGATT
CACCTTCAATAAGTACGCCATGAACTGGGTCCGCCAGGCTCCAGGAAAGGGTTTGG
AATGGGTTGCTCGCATAAGAAGTAAATATAATAATTATGCAACATATTATGCCGATTC
AGTGAAAGACAGGTTCACCATCTCCAGAGATGATTCAAAAAACACTGCCTATCTACA
AATGAACAACTTGAAAACTGAGGACACTGCCGTGTACTACTGTGTGAGACATGGGA
ACTTCGGTAATAGCTACATATCCTACTGGGCTTACTGGGGCCAAGGGACTCTGGTC
ACCGTCTCCTCAGGTGGTGGTGGTTCTGGCGGCGGCGGCTCCGGTGGTGGTGGTT
CTCAGACTGTTGTGACTCAGGAACCTTCACTCACCGTATCACCTGGTGGAACAGTC
ACACTCACTTGTGGCTCCTCGACTGGGGCTGTTACATCTGGCAACTACCCAAACTG
GGTCCAACAAAAACCAGGTCAGGCACCCCGTGGTCTAATAGGTGGGACTAAGTTCC
TCGCCCCCGGTACTCCTGCCAGATTCTCAGGCTCCCTGCTTGGAGGCAAGGCTGC
CCTCACCCTCTCAGGGGTACAGCCAGAGGATGAGGCAGAATATTACTGTGTTCTAT
GGTACAGCAACCGCTGGGTGTTCGGTGGAGGAACCAAACTGACTGTCCTA 1033.
huPSMArat140- artificial nt
ATGTGGAATCTGCTTCACGAAACAGACTCGGCTGTCGCCACCGCGCGGCGCCCGC 169
GGTGGCTGTGCGCCGGGGCGCTGGTCCTGGCGGGTGGATTCTTTCTCCTGGGCTT
CCTCTTTGGGTGGTTTATCAAATCCTCCAACGAAGCTACTAATATTACTCCAAAACAT
AATATGAAGGCATTTTTGGACGAATTGAAAGCCGAGAACATCAAAAAGTTCTTATAC
AATTTTACCCAGATACCACACTTAGCAGGAACCGAACAAAACTTCCAGCTTGCAAAA
CAAATTCAATCTCAGTGGAAAGAGTTTGGCCTGGACTCTGTTGAGCTGGCACATTAT
GACGTCCTGTTGTCTTACCCAAATAAAACTCATCCCAATTACATCTCAATCATTAATG
AAGACGGAAATGAGATCTTCAAAACATCATTAGCTGAACTGTCACCCCCGGGATATG
AGAACATATCAGATGTAGTGCCACCATACAGTGCCTTCTCTCCACAAGGGACACCC
GAGGGGGACCTAGTGTATGTGAACTATGCACGGACTGAAGACTTTTTTAAATTGGA
GCGGGACATGAAGATCAATTGCTCCGGGAAAATTGTGATTGCCAGATACGGGAAAG
TTTTTAGAGGAAATAAAGTTAAAAATGCTCAGCTGGCAGGCGCCAAAGGAGTGATTC
TCTACTCTGACCCTGCTGATTACTTTGCTCCCGGGGTGAAGTCATATCCAGATGGCT
GGAATCTTCCCGGAGGTGGTGTGCAGCGTGGAAACATCCTAAATCTCAATGGTGCA
GGCGACCCTCTCACCCCAGGTTACCCCGCAAATGAATACGCTTATAGGCGGGGAAT
TGCAGAAGCTGTTGGTCTGCCAAGTATTCCAGTTCATCCAATCGGATACTATGACGC
ACAGAAGCTGCTAGAAAAGATGGGTGGCTCCGCACCACCAGACAGCAGCTGGAGG
GGAAGTCTCAAGGTGCCCTACAACGTTGGACCTGGATTTACTGGAAATTTTTCTACA
CAGAAAGTCAAAATGCACATCCATTCTACCAATGAGGTGACAAGAATCTACAATGTG
ATCGGTACTCTCAGGGGAGCAGTGGAGCCAGACAGGTATGTCATTCTCGGAGGTC
ACCGCGACTCATGGGTCTTTGGTGGTATCGACCCTCAGAGCGGAGCAGCTGTGGT
TCATGAAATCGTGAGGAGCTTCGGAACACTGAAGAAGGAAGGCTGGAGACCTAGG
AGAACAATCTTGTTTGCAAGTTGGGATGCAGAGGAATTTGGTCTGCTTGGTTCTACC
GAGTGGGCAGAAGAGAACTCAAGACTCCTGCAAGAGCGTGGAGTGGCTTATATCAA
TGCTGACTCCTCTATAGAAGGCAACTACACCCTGAGAGTTGACTGTACACCCCTGAT
GTACAGTTTGGTACACAATCTAACAAAAGAACTGAAAAGCCCCGATGAAGGCTTCGA
AGGCAAATCCCTTTATGAAAGCTGGACTAAAAAGAGTCCTTCCCCTGAGTTCAGTGG
AATGCCCAGGATCAGCAAATTGGGCTCTGGAAATGACTTTGAGGTGTTTTTCCAACG
ACTGGGAATTGCTTCCGGCAGAGCACGCTATACTAAAAACTGGGAAACAAATAAATT
CAGTGGCTATCCCCTGTATCACAGCGTCTATGAAACCTATGAGTTGGTCGAAAAGTT
TTACGATCCAATGTTCAAATATCACCTGACTGTGGCTCAGGTTCGAGGCGGGATGG
TGTTCGAGCTAGCCAACTCCATAGTGCTGCCTTTTGATTGCCGAGATTATGCCGTAG
TTTTAAGGAAGTATGCTGATAAAATCTACAGCATTTCTATGAAGCATCCACAGGAGA
TGAAGACATATAGTGTATCATTCGATTCACTTTTCTCTGCAGTGAAGAATTTTACCGA
AATTGCTTCTAAGTTTAGTGAGAGGCTCCAGGACTTCGACAAAAGCAATCCAATAGT
ATTGAGAATGATGAACGATCAACTGATGTTTCTGGAGAGAGCATTTATCGATCCATT
AGGCTTACCAGACCGCCCTTTTTATAGACATGTCATCTACGCTCCAAGCAGTCACAA
CAAGTACGCAGGGGAGTCCTTCCCAGGAATCTATGATGCCCTGTTTGACATTGAAA
GCAAGGTGGACCCTTCTAAGGCCTGGGGCGAAGTGAAGAGGCAGATTTATGTGGC
AGCCTTCACCGTGCAGGCAGCCGCAGAGACCTTGAGTGAGGTAGCCTCCGGGGAT
TACAAGGACGACGATGACAAGTAA 1034. huPSMArat140- artificial aa
MWNLLHETDSAVATARRPRWLCAGALVLAGGFFLLGFLFGWFIKSSNEATNITPKHNM 169
KAFLDELKAENIKKFLYNFTQIPHLAGTEQNFQLAKQIQSQWKEFGLDSVELAHYDVLLS
YPNKTHPNYISIINEDGNEIFKTSLAELSPPGYENISDVVPPYSAFSPQGTPEGDLVYVNY
ARTEDFFKLERDMKINCSGKIVIARYGKVFRGNKVKNAQLAGAKGVILYSDPADYFAPG
VKSYPDGWNLPGGGVQRGNILNLNGAGDPLTPGYPANEYAYRRGIAEAVGLPSIPVHPI
GYYDAQKLLEKMGGSAPPDSSWRGSLKVPYNVGPGFTGNFSTQKVKMHIHSTNEVTRI
YNVIGTLRGAVEPDRYVILGGHRDSWVFGGIDPQSGAAVVHEIVRSFGTLKKEGWRPR
RTILFASWDAEEFGLLGSTEWAEENSRLLQERGVAYINADSSIEGNYTLRVDCTPLMYS
LVHNLTKELKSPDEGFEGKSLYESWTKKSPSPEFSGMPRISKLGSGNDFEVFFQRLGIA
SGRARYTKNWETNKFSGYPLYHSVYETYELVEKFYDPMFKYHLTVAQVRGGMVFELA
NSIVLPFDCRDYAVVLRKYADKIYSISMKHPQEMKTYSVSFDSLFSAVKNFTEIASKFSE
RLQDFDKSNPIVLRMMNDQLMFLERAFIDPLGLPDRPFYRHVIYAPSSHNKYAGESFPGI
YDALFDIESKVDPSKAWGEVKRQIYVAAFTVQAAAETLSEVASGDYKDDDDK 1035.
huPSMArat191- artificial nt
ATGTGGAATCTGCTTCACGAAACAGACTCGGCTGTCGCCACCGCGCGGCGCCCGC 258
GGTGGCTGTGCGCCGGGGCGCTGGTCCTGGCGGGTGGATTCTTTCTCCTGGGCTT
CCTCTTTGGGTGGTTTATCAAATCCTCCAACGAAGCTACTAATATTACTCCAAAACAT
AATATGAAGGCATTTTTGGACGAATTGAAAGCCGAGAACATCAAAAAGTTCTTATAC
AATTTTACCCAGATACCACACTTAGCAGGAACCGAACAAAACTTCCAGCTTGCAAAA
CAAATTCAATCTCAGTGGAAAGAGTTTGGCCTGGACTCTGTTGAGCTGGCACATTAT
GACGTCCTGTTGTCTTACCCAAATAAAACTCATCCCAATTACATCTCAATCATTAATG
AAGACGGAAATGAGATCTTCAACACATCCTTATTTGAACCCCCTCCTCCAGGCTATG
AAAATGTGTCGGATATTGTGCCACCTTTCAGCGCTTTCTCTCCCCAAGGAATGCCCG
AGGGCGACCTAGTGTATGTGAACTATGCGCGCACTGAAGACTTTTTTAAATTGGAG
CGGGTCATGAAGATCAATTGTTCTGGGAAGATTGTCATCGCCAGATATGGCCAAGT
GTTCAGAGGAAACAAGGTTAAAAACGCTCAGCTGGCAGGTGCAAAAGGAATCATTC
TGTACTCAGACCCTGCTGATTACTTTGTTCCTGGCGTGAAGTCCTACCCAGATGGCT
GGAACCTCCCTGGAGGTGGCGTTCAGCGTGGAAACGTCCTAAATCTCAATGGTGCA
GGCGACCCGTTAACCCCAGGTTACCCCGCAAATGAATACGCTTATAGGCGGGGAAT
TGCAGAAGCTGTTGGTCTGCCAAGTATTCCAGTTCATCCAATCGGATACTATGACGC
ACAGAAGCTGCTAGAAAAGATGGGTGGCTCCGCACCACCAGACAGCAGCTGGAGG
GGAAGCTTGAAGGTGCCCTACAACGTTGGACCTGGATTTACTGGAAATTTTTCTACA
CAGAAAGTCAAAATGCACATCCATTCTACCAATGAGGTGACAAGAATCTACAATGTG
ATCGGTACTCTCAGGGGAGCAGTGGAGCCAGACAGGTATGTCATTCTCGGAGGTC
ACCGCGACTCATGGGTCTTTGGTGGTATCGACCCTCAGAGCGGAGCAGCTGTGGT
TCATGAAATCGTGAGGAGCTTCGGAACACTGAAGAAGGAAGGCTGGAGACCTAGG
AGAACAATCTTGTTTGCAAGTTGGGATGCAGAGGAATTTGGTCTGCTTGGTTCTACC
GAGTGGGCAGAAGAGAACTCAAGACTCCTGCAAGAGCGTGGAGTGGCTTATATCAA
TGCTGACTCCTCTATAGAAGGCAACTACACCCTGAGAGTTGACTGTACACCCCTGAT
GTACAGTTTGGTACACAATCTAACAAAAGAACTGAAAAGCCCCGATGAAGGCTTCGA
AGGCAAATCCCTTTATGAAAGCTGGACTAAAAAGAGTCCTTCCCCTGAGTTCAGTGG
AATGCCCAGGATCAGCAAATTGGGCTCTGGAAATGACTTTGAGGTGTTTTTCCAACG
ACTGGGAATTGCTTCCGGCAGAGCACGCTATACTAAAAACTGGGAAACAAATAAATT
CAGTGGCTATCCCCTGTATCACAGCGTCTATGAAACCTATGAGTTGGTCGAAAAGTT
TTACGATCCAATGTTCAAATATCACCTGACTGTGGCTCAGGTTCGAGGCGGGATGG
TGTTCGAGCTAGCCAACTCCATAGTGCTGCCTTTTGATTGCCGAGATTATGCCGTAG
TTTTAAGGAAGTATGCTGATAAAATCTACAGCATTTCTATGAAGCATCCACAGGAGA
TGAAGACATATAGTGTATCATTCGATTCACTTTTCTCTGCAGTGAAGAATTTTACCGA
AATTGCTTCTAAGTTTAGTGAGAGGCTCCAGGACTTCGACAAAAGCAATCCAATAGT
ATTGAGAATGATGAACGATCAACTGATGTTTCTGGAGAGAGCATTTATCGATCCATT
AGGCTTACCAGACCGCCCTTTTTATAGACATGTCATCTACGCTCCAAGCAGTCACAA
CAAGTACGCAGGGGAGTCCTTCCCAGGAATCTATGATGCCCTGTTTGACATTGAAA
GCAAGGTGGACCCTTCTAAGGCCTGGGGCGAAGTGAAGAGGCAGATTTATGTGGC
AGCCTTCACCGTGCAGGCAGCCGCAGAGACCTTGAGTGAGGTAGCCTCCGGGGAT
TACAAGGACGACGATGACAAGTAA 1036. huPSMArat191- artificial aa
MWNLLHETDSAVATARRPRWLCAGALVLAGGFFLLGFLFGWFIKSSNEATNITPKHNM 258
KAFLDELKAENIKKFLYNFTQIPHLAGTEQNFQLAKQIQSQWKEFGLDSVELAHYDVLLS
YPNKTHPNYISIINEDGNEIFNTSLFEPPPPGYENVSDIVPPFSAFSPQGMPEGDLVYVN
YARTEDFFKLERVMKINCSGKIVIARYGQVFRGNKVKNAQLAGAKGIILYSDPADYFVPG
VKSYPDGWNLPGGGVQRGNVLNLNGAGDPLTPGYPANEYAYRRGIAEAVGLPSIPVHP
IGYYDAQKLLEKMGGSAPPDSSWRGSLKVPYNVGPGFTGNFSTQKVKMHIHSTNEVTR
IYNVIGTLRGAVEPDRYVILGGHRDSWVFGGIDPQSGAAVVHEIVRSFGTLKKEGWRPR
RTILFASWDAEEFGLLGSTEWAEENSRLLQERGVAYINADSSIEGNYTLRVDCTPLMYS
LVHNLTKELKSPDEGFEGKSLYESWTKKSPSPEFSGMPRISKLGSGNDFEVFFQRLGIA
SGRARYTKNWETNKFSGYPLYHSVYETYELVEKFYDPMFKYHLTVAQVRGGMVFELA
NSIVLPFDCRDYAVVLRKYADKIYSISMKHPQEMKTYSVSFDSLFSAVKNFTEIASKFSE
RLQDFDKSNPIVLRMMNDQLMFLERAFIDPLGLPDRPFYRHVIYAPSSHNKYAGESFPGI
YDALFDIESKVDPSKAWGEVKRQIYVAAFTVQAAAETLSEVASGDYKDDDDK 1037.
huPSMArat281- artificial nt
ATGTGGAATCTGCTTCACGAAACAGACTCGGCTGTCGCCACCGCGCGGCGCCCGC 284
GGTGGCTGTGCGCCGGGGCGCTGGTCCTGGCGGGTGGATTCTTTCTCCTGGGCTT
CCTCTTTGGGTGGTTTATCAAATCCTCCAACGAAGCTACTAATATTACTCCAAAACAT
AATATGAAGGCATTTTTGGACGAATTGAAAGCCGAGAACATCAAAAAGTTCTTATAC
AATTTTACCCAGATACCACACTTAGCAGGAACCGAACAAAACTTCCAGCTTGCAAAA
CAAATTCAATCTCAGTGGAAAGAGTTTGGCCTGGACTCTGTTGAGCTGGCACATTAT
GACGTCCTGTTGTCTTACCCAAATAAAACTCATCCCAATTACATCTCAATCATTAATG
AAGACGGAAATGAGATCTTCAACACATCCTTATTTGAACCCCCTCCTCCAGGCTATG
AAAATGTGTCGGATATTGTGCCACCTTTCAGCGCTTTCTCTCCCCAAGGAATGCCCG
AGGGCGACCTAGTGTATGTGAACTATGCACGGACTGAAGACTTTTTTAAATTGGAGC
GGGACATGAAGATCAATTGCTCCGGGAAAATTGTGATTGCCAGATACGGGAAAGTT
TTTAGAGGAAATAAAGTTAAAAATGCTCAGCTGGCAGGCGCCAAAGGAGTGATTCT
CTACTCTGACCCTGCTGATTACTTTGCTCCCGGGGTGAAGTCATATCCAGATGGCT
GGAATCTTCCCGGAGGTGGTGTGCAGCGTGGAAACATCCTAAATCTCAATGGTGCA
GGCGACCCGTTAACCCCAGGTTACCCCGCAAATGAATACGCTTATAGGCATGAGTT
CACAGAAGCTGTTGGTCTGCCAAGTATTCCAGTTCATCCAATCGGATACTATGACGC
ACAGAAGCTGCTAGAAAAGATGGGTGGCTCCGCACCACCAGACAGCAGCTGGAGG
GGAAGCTTGAAGGTGCCCTACAACGTTGGACCTGGATTTACTGGAAATTTTTCTACA
CAGAAAGTCAAAATGCACATCCATTCTACCAATGAGGTGACAAGAATCTACAATGTG
ATCGGTACTCTCAGGGGAGCAGTGGAGCCAGACCGGTATGTCATTCTCGGAGGTC
ACCGCGACTCATGGGTCTTTGGTGGTATCGACCCTCAGAGCGGAGCAGCTGTGGT
TCATGAAATCGTGAGGAGCTTCGGAACACTGAAGAAGGAAGGCTGGAGACCTAGG
AGAACAATCTTGTTTGCAAGTTGGGATGCAGAGGAATTTGGTCTGCTTGGTTCTACC
GAGTGGGCAGAAGAGAACTCAAGACTCCTGCAAGAGCGTGGAGTGGCTTATATCAA
TGCTGACTCCTCTATAGAAGGCAACTACACCCTGAGAGTTGACTGTACACCCCTGAT
GTACAGTTTGGTACACAATCTAACAAAAGAACTGAAAAGCCCCGATGAAGGCTTCGA
AGGCAAATCCCTTTATGAAAGCTGGACTAAAAAGAGTCCTTCCCCTGAGTTCAGTGG
AATGCCCAGGATCAGCAAATTGGGCTCTGGAAATGACTTTGAGGTGTTTTTCCAACG
ACTGGGAATTGCTTCCGGCAGAGCACGCTATACTAAAAACTGGGAAACAAATAAATT
CAGTGGCTATCCCCTGTATCACAGCGTCTATGAAACCTATGAGTTGGTCGAAAAGTT
TTACGATCCAATGTTCAAATATCACCTGACTGTGGCTCAGGTTCGAGGCGGGATGG
TGTTCGAGCTAGCCAACTCCATAGTGCTGCCTTTTGATTGCCGAGATTATGCCGTAG
TTTTAAGGAAGTATGCTGATAAAATCTACAGCATTTCTATGAAGCATCCACAGGAGA
TGAAGACATATAGTGTATCATTCGATTCACTTTTCTCTGCAGTGAAGAATTTTACCGA
AATTGCTTCTAAGTTTAGTGAGAGGCTCCAGGACTTCGACAAAAGCAATCCAATAGT
ATTGAGAATGATGAACGATCAACTGATGTTTCTGGAGAGAGCATTTATCGATCCATT
AGGCTTACCAGACCGCCCTTTTTATAGACATGTCATCTACGCTCCAAGCAGTCACAA
CAAGTACGCAGGGGAGTCCTTCCCAGGAATCTATGATGCCCTGTTTGACATTGAAA
GCAAGGTGGACCCTTCTAAGGCCTGGGGCGAAGTGAAGAGGCAGATTTATGTGGC
AGCCTTCACCGTGCAGGCAGCCGCAGAGACCTTGAGTGAGGTAGCCTCCGGGGAT
TACAAGGACGACGATGACAAGTAA 1038. huPSMArat281- artificial aa
MWNLLHETDSAVATARRPRWLCAGALVLAGGFFLLGFLFGWFIKSSNEATNITPKHNM 284
KAFLDELKAENIKKFLYNFTQIPHLAGTEQNFQLAKQIQSQWKEFGLDSVELAHYDVLLS
YPNKTHPNYISIINEDGNEIFNTSLFEPPPPGYENVSDIVPPFSAFSPQGMPEGDLVYVN
YARTEDFFKLERDMKINCSGKIVIARYGKVFRGNKVKNAQLAGAKGVILYSDPADYFAP
GVKSYPDGWNLPGGGVQRGNILNLNGAGDPLTPGYPANEYAYRHEFTEAVGLPSIPVH
PIGYYDAQKLLEKMGGSAPPDSSWRGSLKVPYNVGPGFTGNFSTQKVKMHIHSTNEVT
RIYNVIGTLRGAVEPDRYVILGGHRDSWVFGGIDPQSGAAVVHEIVRSFGTLKKEGWRP
RRTILFASWDAEEFGLLGSTEWAEENSRLLQERGVAYINADSSIEGNYTLRVDCTPLMY
SLVHNLTKELKSPDEGFEGKSLYESWTKKSPSPEFSGMPRISKLGSGNDFEVFFQRLGI
ASGRARYTKNWETNKFSGYPLYHSVYETYELVEKFYDPMFKYHLTVAQVRGGMVFELA
NSIVLPFDCRDYAVVLRKYADKIYSISMKHPQEMKTYSVSFDSLFSAVKNFTEIASKFSE
RLQDFDKSNPIVLRMMNDQLMFLERAFIDPLGLPDRPFYRHVIYAPSSHNKYAGESFPGI
YDALFDIESKVDPSKAWGEVKRQIYVAAFTVQAAAETLSEVASGDYKDDDDK 1039.
huPSMArat300- artificial nt
ATGTGGAATCTGCTTCACGAAACAGACTCGGCTGTCGCCACCGCGCGGCGCCCGC 344
GGTGGCTGTGCGCCGGGGCGCTGGTCCTGGCGGGTGGATTCTTTCTCCTGGGCTT
CCTCTTTGGGTGGTTTATCAAATCCTCCAACGAAGCTACTAATATTACTCCAAAACAT
AATATGAAGGCATTTTTGGACGAATTGAAAGCCGAGAACATCAAAAAGTTCTTATAC
AATTTTACCCAGATACCACACTTAGCAGGAACCGAACAAAACTTCCAGCTTGCAAAA
CAAATTCAATCTCAGTGGAAAGAGTTTGGCCTGGACTCTGTTGAGCTGGCACATTAT
GACGTCCTGTTGTCTTACCCAAATAAAACTCATCCCAATTACATCTCAATCATTAATG
AAGACGGAAATGAGATCTTCAACACATCCTTATTTGAACCCCCTCCTCCAGGCTATG
AAAATGTGTCGGATATTGTGCCACCTTTCAGCGCTTTCTCTCCCCAAGGAATGCCCG
AGGGCGACCTAGTGTATGTGAACTATGCACGGACTGAAGACTTTTTTAAATTGGAGC
GGGACATGAAGATCAATTGCTCCGGGAAAATTGTGATTGCCAGATACGGGAAAGTT
TTTAGAGGAAATAAAGTTAAAAATGCTCAGCTGGCAGGCGCCAAAGGAGTGATTCT
CTACTCTGACCCTGCTGATTACTTTGCTCCCGGGGTGAAGTCATATCCAGATGGCT
GGAATCTTCCCGGAGGTGGTGTGCAGCGTGGAAACATCCTAAATCTCAATGGTGCA
GGCGACCCGTTAACCCCAGGTTACCCCGCAAATGAATACGCTTATAGGCGGGGAAT
TGCAGAAGCTGTTGGTCTGCCAAGTATTCCAGTTCATCCAATCGGATACGATGATGC
CCAGAAACTATTAGAACATATGGGTGGCTCCGCACCCCCTGACAGCAGCTGGAAGG
GAGGACTAAAAGTGCCTTACAACGTGGGACCTGGCTTCGCTGGAAACTTCTCAAAA
CAAAAGGTCAAGCTGCACATCCATTCTACCAATGAGGTGACAAGAATCTACAATGTG
ATCGGTACTCTCAGGGGAGCAGTGGAGCCAGACCGGTATGTCATTCTCGGAGGTC
ACCGCGACTCATGGGTCTTTGGTGGTATCGACCCTCAGAGCGGAGCAGCTGTGGT
TCATGAAATCGTGAGGAGCTTCGGAACACTGAAGAAGGAAGGCTGGAGACCTAGG
AGAACAATCTTGTTTGCAAGTTGGGATGCAGAGGAATTTGGTCTGCTTGGTTCTACC
GAGTGGGCAGAAGAGAACTCAAGACTCCTGCAAGAGCGTGGAGTGGCTTATATCAA
TGCTGACTCCTCTATAGAAGGCAACTACACCCTGAGAGTTGACTGTACACCCCTGAT
GTACAGTTTGGTACACAATCTAACAAAAGAACTGAAAAGCCCCGATGAAGGCTTCGA
AGGCAAATCCCTTTATGAAAGCTGGACTAAAAAGAGTCCTTCCCCTGAGTTCAGTGG
AATGCCCAGGATCAGCAAATTGGGCTCTGGAAATGACTTTGAGGTGTTTTTCCAACG
ACTGGGAATTGCTTCCGGCAGAGCACGCTATACTAAAAACTGGGAAACAAATAAATT
CAGTGGCTATCCCCTGTATCACAGCGTCTATGAAACCTATGAGTTGGTCGAAAAGTT
TTACGATCCAATGTTCAAATATCACCTGACTGTGGCTCAGGTTCGAGGCGGGATGG
TGTTCGAGCTAGCCAACTCCATAGTGCTGCCTTTTGATTGCCGAGATTATGCCGTAG
TTTTAAGGAAGTATGCTGATAAAATCTACAGCATTTCTATGAAGCATCCACAGGAGA
TGAAGACATATAGTGTATCATTCGATTCACTTTTCTCTGCAGTGAAGAATTTTACCGA
AATTGCTTCTAAGTTTAGTGAGAGGCTCCAGGACTTCGACAAAAGCAATCCAATAGT
ATTGAGAATGATGAACGATCAACTGATGTTTCTGGAGAGAGCATTTATCGATCCATT
AGGCTTACCAGACCGCCCTTTTTATAGACATGTCATCTACGCTCCAAGCAGTCACAA
CAAGTACGCAGGGGAGTCCTTCCCAGGAATCTATGATGCCCTGTTTGACATTGAAA
GCAAGGTGGACCCTTCTAAGGCCTGGGGCGAAGTGAAGAGGCAGATTTATGTGGC
AGCCTTCACCGTGCAGGCAGCCGCAGAGACCTTGAGTGAGGTAGCCTCCGGGGAT
TACAAGGACGACGATGACAAGTAA 1040. huPSMArat300- artificial aa
MWNLLHETDSAVATARRPRWLCAGALVLAGGFFLLGFLFGWFIKSSNEATNITPKHNM 344
KAFLDELKAENIKKFLYNFTQIPHLAGTEQNFQLAKQIQSQWKEFGLDSVELAHYDVLLS
YPNKTHPNYISIINEDGNEIFNTSLFEPPPPGYENVSDIVPPFSAFSPQGMPEGDLVYVN
YARTEDFFKLERDMKINCSGKIVIARYGKVFRGNKVKNAQLAGAKGVILYSDPADYFAP
GVKSYPDGWNLPGGGVQRGNILNLNGAGDPLTPGYPANEYAYRRGIAEAVGLPSIPVH
PIGYDDAQKLLEHMGGSAPPDSSWKGGLKVPYNVGPGFAGNFSKQKVKLHIHSTNEVT
RIYNVIGTLRGAVEPDRYVILGGHRDSWVFGGIDPQSGAAVVHEIVRSFGTLKKEGWRP
RRTILFASWDAEEFGLLGSTEWAEENSRLLQERGVAYINADSSIEGNYTLRVDCTPLMY
SLVHNLTKELKSPDEGFEGKSLYESWTKKSPSPEFSGMPRISKLGSGNDFEVFFQRLGI
ASGRARYTKNWETNKFSGYPLYHSVYETYELVEKFYDPMFKYHLTVAQVRGGMVFELA
NSIVLPFDCRDYAVVLRKYADKIYSISMKHPQEMKTYSVSFDSLFSAVKNFTEIASKFSE
RLQDFDKSNPIVLRMMNDQLMFLERAFIDPLGLPDRPFYRHVIYAPSSHNKYAGESFPGI
YDALFDIESKVDPSKAWGEVKRQIYVAAFTVQAAAETLSEVASGDYKDDDDK 1041.
huPSMArat598- artificial nt
ATGTGGAATCTGCTTCACGAAACAGACTCGGCTGTCGCCACCGCGCGGCGCCCGC 617
GGTGGCTGTGCGCCGGGGCGCTGGTCCTGGCGGGTGGATTCTTTCTCCTGGGCTT
CCTCTTTGGGTGGTTTATCAAATCCTCCAACGAAGCTACTAATATTACTCCAAAACAT
AATATGAAGGCATTTTTGGACGAATTGAAAGCCGAGAACATCAAAAAGTTCTTATAC
AATTTTACCCAGATACCACACTTAGCAGGAACCGAACAAAACTTCCAGCTTGCAAAA
CAAATTCAATCTCAGTGGAAAGAGTTTGGCCTGGACTCTGTTGAGCTGGCACATTAT
GACGTCCTGTTGTCTTACCCAAATAAAACTCATCCCAATTACATCTCAATCATTAATG
AAGACGGAAATGAGATCTTCAACACATCCTTATTTGAACCCCCTCCTCCAGGCTATG
AAAATGTGTCGGATATTGTGCCACCTTTCAGCGCTTTCTCTCCCCAAGGAATGCCCG
AGGGCGACCTAGTGTATGTGAACTATGCACGGACTGAAGACTTTTTTAAATTGGAGC
GGGACATGAAGATCAATTGCTCCGGGAAAATTGTGATTGCCAGATACGGGAAAGTT
TTTAGAGGAAATAAAGTTAAAAATGCTCAGCTGGCAGGCGCCAAAGGAGTGATTCT
CTACTCTGACCCTGCTGATTACTTTGCTCCCGGGGTGAAGTCATATCCAGATGGCT
GGAATCTTCCCGGAGGTGGTGTGCAGCGTGGAAACATCCTAAATCTCAATGGTGCA
GGCGACCCTCTCACCCCAGGTTACCCCGCAAATGAATACGCTTATAGGCGGGGAAT
TGCAGAAGCTGTTGGTCTGCCAAGTATTCCAGTTCATCCAATCGGATACTATGACGC
ACAGAAGCTGCTAGAAAAGATGGGTGGCTCCGCACCACCAGACAGCAGCTGGAGG
GGAAGTCTCAAGGTGCCCTACAACGTTGGACCTGGATTTACTGGAAATTTTTCTACA
CAGAAAGTCAAAATGCACATCCATTCTACCAATGAGGTGACAAGAATCTACAATGTG
ATCGGTACTCTCAGGGGAGCAGTGGAGCCAGACAGGTATGTCATTCTCGGAGGTC
ACCGCGACTCATGGGTCTTTGGTGGTATCGACCCTCAGAGCGGAGCAGCTGTGGT
TCATGAAATCGTGAGGAGCTTCGGAACACTGAAGAAGGAAGGCTGGAGACCTAGG
AGAACAATCTTGTTTGCAAGTTGGGATGCAGAGGAATTTGGTCTGCTTGGTTCTACC
GAGTGGGCAGAAGAGAACTCAAGACTCCTGCAAGAGCGTGGAGTGGCTTATATCAA
TGCTGACTCCTCTATAGAAGGCAACTACACCCTGAGAGTTGACTGTACACCCCTGAT
GTACAGTTTGGTACACAATCTAACAAAAGAACTGAAAAGCCCCGATGAAGGCTTCGA
AGGCAAATCCCTTTATGAAAGCTGGACTAAAAAGAGTCCTTCCCCTGAGTTCAGTGG
AATGCCCAGGATCAGCAAATTGGGCTCTGGAAATGACTTTGAGGTGTTTTTCCAACG
ACTGGGAATTGCTTCCGGCAGAGCACGCTATACTAAAAACTGGGAAACAAATAAATT
CAGTGGCTATCCCCTGTATCACAGCGTCTATGAAACCTATGAGTTGGTCGAAAAGTT
TTACGATCCAATGTTCAAATATCACCTGACTGTGGCTCAGGTTCGAGGCGGGATGG
TGTTCGAGCTCGCCAACTCCATAGTGCTGCCTTTTGATTGCCAAAGTTATGCTGTAG
CTCTGAAGAAACATGCTGAGACTATCTACAACATTTCAATGAATCATCCACAGGAGA
TGAAGACATATAGTGTAAGCTTCGATTCACTTTTCTCTGCAGTGAAGAATTTTACCGA
AATTGCTTCTAAGTTTAGTGAGAGGCTCCAGGACTTCGACAAAAGCAATCCAATAGT
ATTGAGAATGATGAACGATCAACTGATGTTTCTGGAGAGAGCATTTATCGATCCATT
AGGCTTACCAGACCGCCCTTTTTATAGACATGTCATCTACGCTCCAAGCAGTCACAA
CAAGTACGCAGGGGAGTCCTTCCCAGGAATCTATGACGCGTTGTTTGACATTGAAA
GCAAGGTGGACCCTTCTAAGGCCTGGGGCGAAGTGAAGAGGCAGATTTATGTGGC
AGCCTTCACCGTGCAGGCAGCCGCAGAGACCTTGAGTGAGGTAGCCTCCGGGGAT
TACAAGGACGACGATGACAAGTAA 1042. huPSMArat598- artificial aa
MWNLLHETDSAVATARRPRWLCAGALVLAGGFFLLGFLFGWFIKSSNEATNITPKHNM 617
KAFLDELKAENIKKFLYNFTQIPHLAGTEQNFQLAKQIQSQWKEFGLDSVELAHYDVLLS
YPNKTHPNYISIINEDGNEIFNTSLFEPPPPGYENVSDIVPPFSAFSPQGMPEGDLVYVN
YARTEDFFKLERDMKINCSGKIVIARYGKVFRGNKVKNAQLAGAKGVILYSDPADYFAP
GVKSYPDGWNLPGGGVQRGNILNLNGAGDPLTPGYPANEYAYRRGIAEAVGLPSIPVH
PIGYYDAQKLLEKMGGSAPPDSSWRGSLKVPYNVGPGFTGNFSTQKVKMHIHSTNEVT
RIYNVIGTLRGAVEPDRYVILGGHRDSWVFGGIDPQSGAAVVHEIVRSFGTLKKEGWRP
RRTILFASWDAEEFGLLGSTEWAEENSRLLQERGVAYINADSSIEGNYTLRVDCTPLMY
SLVHNLTKELKSPDEGFEGKSLYESWTKKSPSPEFSGMPRISKLGSGNDFEVFFQRLGI
ASGRARYTKNWETNKFSGYPLYHSVYETYELVEKFYDPMFKYHLTVAQVRGGMVFELA
NSIVLPFDCQSYAVALKKHAETIYNISMNHPQEMKTYSVSFDSLFSAVKNFTEIASKFSE
RLQDFDKSNPIVLRMMNDQLMFLERAFIDPLGLPDRPFYRHVIYAPSSHNKYAGESFPGI
YDALFDIESKVDPSKAWGEVKRQIYVAAFTVQAAAETLSEVASGDYKDDDDK 1043.
huPSMArat683- artificial nt
ATGTGGAATCTGCTTCACGAAACAGACTCGGCTGTCGCCACCGCGCGGCGCCCGC 690
GGTGGCTGTGCGCCGGGGCGCTGGTCCTGGCGGGTGGATTCTTTCTCCTGGGCTT
CCTCTTTGGGTGGTTTATCAAATCCTCCAACGAAGCTACTAATATTACTCCAAAACAT
AATATGAAGGCATTTTTGGACGAATTGAAAGCCGAGAACATCAAAAAGTTCTTATAC
AATTTTACCCAGATACCACACTTAGCAGGAACCGAACAAAACTTCCAGCTTGCAAAA
CAAATTCAATCTCAGTGGAAAGAGTTTGGCCTGGACTCTGTTGAGCTGGCACATTAT
GACGTCCTGTTGTCTTACCCAAATAAAACTCATCCCAATTACATCTCAATCATTAATG
AAGACGGAAATGAGATCTTCAACACATCCTTATTTGAACCCCCTCCTCCAGGCTATG
AAAATGTGTCGGATATTGTGCCACCTTTCAGCGCTTTCTCTCCCCAAGGAATGCCCG
AGGGCGACCTAGTGTATGTGAACTATGCACGGACTGAAGACTTTTTTAAATTGGAGC
GGGACATGAAGATCAATTGCTCCGGGAAAATTGTGATTGCCAGATACGGGAAAGTT
TTTAGAGGAAATAAAGTTAAAAATGCTCAGCTGGCAGGCGCCAAAGGAGTGATTCT
CTACTCTGACCCTGCTGATTACTTTGCTCCCGGGGTGAAGTCATATCCAGATGGCT
GGAATCTTCCCGGAGGTGGTGTGCAGCGTGGAAACATCCTAAATCTCAATGGTGCA
GGCGACCCTCTCACCCCAGGTTACCCCGCAAATGAATACGCTTATAGGCGGGGAAT
TGCAGAAGCTGTTGGTCTGCCAAGTATTCCAGTTCATCCAATCGGATACTATGACGC
ACAGAAGCTGCTAGAAAAGATGGGTGGCTCCGCACCACCAGACAGCAGCTGGAGG
GGAAGTCTCAAGGTGCCCTACAACGTTGGACCTGGATTTACTGGAAATTTTTCTACA
CAGAAAGTCAAAATGCACATCCATTCTACCAATGAGGTGACAAGAATCTACAATGTG
ATCGGTACTCTCAGGGGAGCAGTGGAGCCAGACAGGTATGTCATTCTCGGAGGTC
ACCGCGACTCATGGGTCTTTGGTGGTATCGACCCTCAGAGCGGAGCAGCTGTGGT
TCATGAAATCGTGAGGAGCTTCGGAACACTGAAGAAGGAAGGCTGGAGACCTAGG
AGAACAATCTTGTTTGCAAGTTGGGATGCAGAGGAATTTGGTCTGCTTGGTTCTACC
GAGTGGGCAGAAGAGAACTCAAGACTCCTGCAAGAGCGTGGAGTGGCTTATATCAA
TGCTGACTCCTCTATAGAAGGCAACTACACCCTGAGAGTTGACTGTACACCCCTGAT
GTACAGTTTGGTACACAATCTAACAAAAGAACTGAAAAGCCCCGATGAAGGCTTCGA
AGGCAAATCCCTTTATGAAAGCTGGACTAAAAAGAGTCCTTCCCCTGAGTTCAGTGG
AATGCCCAGGATCAGCAAATTGGGCTCTGGAAATGACTTTGAGGTGTTTTTCCAACG
ACTGGGAATTGCTTCCGGCAGAGCACGCTATACTAAAAACTGGGAAACAAATAAATT
CAGTGGCTATCCCCTGTATCACAGCGTCTATGAAACCTATGAGTTGGTCGAAAAGTT
TTACGATCCAATGTTCAAATATCACCTGACTGTGGCTCAGGTTCGAGGCGGGATGG
TGTTCGAGCTCGCCAACTCCATAGTGCTGCCTTTTGATTGCCGAGATTATGCCGTAG
TTTTAAGGAAGTATGCTGATAAAATCTACAGCATTTCTATGAAGCATCCACAGGAGA
TGAAGACATATAGTGTATCATTCGATTCACTTTTCTCTGCAGTGAAGAATTTTACCGA
AATTGCTTCTAAGTTTAGTGAGAGGCTCCAGGACTTCGACAAAAGCAATCCAATAGT
ATTGAGAATGATGAACGATCAACTGATGTTTCTGGAGCGCGCATTTATCGATCCATT
AGGCTTACCAGGAAGGCCTTTCTACAGGCATATCATCTACGCTCCAAGCAGTCACA
ACAAGTACGCAGGGGAGTCCTTCCCAGGAATCTATGACGCGTTGTTTGACATTGAA
AGCAAGGTGGACCCTTCTAAGGCCTGGGGCGAAGTGAAGAGGCAGATTTATGTGG
CAGCCTTCACCGTGCAGGCAGCCGCAGAGACCTTGAGTGAGGTAGCCTCCGGGGA
TTACAAGGACGACGATGACAAGTAA 1044. huPSMArat683- artificial aa
MWNLLHETDSAVATARRPRWLCAGALVLAGGFFLLGFLFGWFIKSSNEATNITPKHNM 690
KAFLDELKAENIKKFLYNFTQIPHLAGTEQNFQLAKQIQSQWKEFGLDSVELAHYDVLLS
YPNKTHPNYISIINEDGNEIFNTSLFEPPPPGYENVSDIVPPFSAFSPQGMPEGDLVYVN
YARTEDFFKLERDMKINCSGKIVIARYGKVFRGNKVKNAQLAGAKGVILYSDPADYFAP
GVKSYPDGWNLPGGGVQRGNILNLNGAGDPLTPGYPANEYAYRRGIAEAVGLPSIPVH
PIGYYDAQKLLEKMGGSAPPDSSWRGSLKVPYNVGPGFTGNFSTQKVKMHIHSTNEVT
RIYNVIGTLRGAVEPDRYVILGGHRDSWVFGGIDPQSGAAVVHEIVRSFGTLKKEGWRP
RRTILFASWDAEEFGLLGSTEWAEENSRLLQERGVAYINADSSIEGNYTLRVDCTPLMY
SLVHNLTKELKSPDEGFEGKSLYESWTKKSPSPEFSGMPRISKLGSGNDFEVFFQRLGI
ASGRARYTKNWETNKFSGYPLYHSVYETYELVEKFYDPMFKYHLTVAQVRGGMVFELA
NSIVLPFDCRDYAVVLRKYADKIYSISMKHPQEMKTYSVSFDSLFSAVKNFTEIASKFSE
RLQDFDKSNPIVLRMMNDQLMFLERAFIDPLGLPGRPFYRHIIYAPSSHNKYAGESFPGI
YDALFDIESKVDPSKAWGEVKRQIYVAAFTVQAAAETLSEVASGDYKDDDDK 1045.
huPSMArat716- artificial nt
ATGTGGAATCTGCTTCACGAAACAGACTCGGCTGTCGCCACCGCGCGGCGCCCGC 750
GGTGGCTGTGCGCCGGGGCGCTGGTCCTGGCGGGTGGATTCTTTCTCCTGGGCTT
CCTCTTTGGGTGGTTTATCAAATCCTCCAACGAAGCTACTAATATTACTCCAAAACAT
AATATGAAGGCATTTTTGGACGAATTGAAAGCCGAGAACATCAAAAAGTTCTTATAC
AATTTTACCCAGATACCACACTTAGCAGGAACCGAACAAAACTTCCAGCTTGCAAAA
CAAATTCAATCTCAGTGGAAAGAGTTTGGCCTGGACTCTGTTGAGCTGGCACATTAT
GACGTCCTGTTGTCTTACCCAAATAAAACTCATCCCAATTACATCTCAATCATTAATG
AAGACGGAAATGAGATCTTCAACACATCCTTATTTGAACCCCCTCCTCCAGGCTATG
AAAATGTGTCGGATATTGTGCCACCTTTCAGCGCTTTCTCTCCCCAAGGAATGCCCG
AGGGCGACCTAGTGTATGTGAACTATGCACGGACTGAAGACTTTTTTAAATTGGAGC
GGGACATGAAGATCAATTGCTCCGGGAAAATTGTGATTGCCAGATACGGGAAAGTT
TTTAGAGGAAATAAAGTTAAAAATGCTCAGCTGGCAGGCGCCAAAGGAGTGATTCT
CTACTCTGACCCTGCTGATTACTTTGCTCCCGGGGTGAAGTCATATCCAGATGGCT
GGAATCTTCCCGGAGGTGGTGTGCAGCGTGGAAACATCCTAAATCTCAATGGTGCA
GGCGACCCTCTCACCCCAGGTTACCCCGCAAATGAATACGCTTATAGGCGGGGAAT
TGCAGAAGCTGTTGGTCTGCCAAGTATTCCAGTTCATCCAATCGGATACTATGACGC
ACAGAAGCTGCTAGAAAAGATGGGTGGCTCCGCACCACCAGACAGCAGCTGGAGG
GGAAGTCTCAAGGTGCCCTACAACGTTGGACCTGGATTTACTGGAAATTTTTCTACA
CAGAAAGTCAAAATGCACATCCATTCTACCAATGAGGTGACAAGAATCTACAATGTG
ATCGGTACTCTCAGGGGAGCAGTGGAGCCAGACAGGTATGTCATTCTCGGAGGTC
ACCGCGACTCATGGGTCTTTGGTGGTATCGACCCTCAGAGCGGAGCAGCTGTGGT
TCATGAAATCGTGAGGAGCTTCGGAACACTGAAGAAGGAAGGCTGGAGACCTAGG
AGAACAATCTTGTTTGCAAGTTGGGATGCAGAGGAATTTGGTCTGCTTGGTTCTACC
GAGTGGGCAGAAGAGAACTCAAGACTCCTGCAAGAGCGTGGAGTGGCTTATATCAA
TGCTGACTCCTCTATAGAAGGCAACTACACCCTGAGAGTTGACTGTACACCCCTGAT
GTACAGTTTGGTACACAATCTAACAAAAGAACTGAAAAGCCCCGATGAAGGCTTCGA
AGGCAAATCCCTTTATGAAAGCTGGACTAAAAAGAGTCCTTCCCCTGAGTTCAGTGG
AATGCCCAGGATCAGCAAATTGGGCTCTGGAAATGACTTTGAGGTGTTTTTCCAACG
ACTGGGAATTGCTTCCGGCAGAGCACGCTATACTAAAAACTGGGAAACAAATAAATT
CAGTGGCTATCCCCTGTATCACAGCGTCTATGAAACCTATGAGTTGGTCGAAAAGTT
TTACGATCCAATGTTCAAATATCACCTGACTGTGGCTCAGGTTCGAGGCGGGATGG
TGTTCGAGCTCGCCAACTCCATAGTGCTGCCTTTTGATTGCCGAGATTATGCCGTAG
TTTTAAGGAAGTATGCTGATAAAATCTACAGCATTTCTATGAAGCATCCACAGGAGA
TGAAGACATATAGTGTATCATTCGATTCACTTTTCTCTGCAGTGAAGAATTTTACCGA
AATTGCTTCTAAGTTTAGTGAGAGGCTCCAGGACTTCGACAAAAGCAATCCAATAGT
ATTGAGAATGATGAACGATCAACTGATGTTTCTGGAGCGCGCATTTATCGATCCATT
AGGCTTACCAGACCGCCCTTTTTATAGACATGTCATCTACGCTCCAAGCAGTCACAA
CAAGTACGCAGGGGAGTCCTTCCCAGGAATCTATGACGCGTTGTTTGACATTAATAA
CAAAGTCGATACTTCTAAGGCCTGGAGAGAAGTGAAAAGACAGATTTCTATTGCAGC
CTTTACAGTGCAAGCTGCAGCAGAGACTCTGAGAGAAGTAGACTCCGGGGATTACA
AGGACGACGATGACAAGTAA 1046. huPSMArat716- artificial aa
MWNLLHETDSAVATARRPRWLCAGALVLAGGFFLLGFLFGWFIKSSNEATNITPKHNM 750
KAFLDELKAENIKKFLYNFTQIPHLAGTEQNFQLAKQIQSQWKEFGLDSVELAHYDVLLS
YPNKTHPNYISIINEDGNEIFNTSLFEPPPPGYENVSDIVPPFSAFSPQGMPEGDLVYVN
YARTEDFFKLERDMKINCSGKIVIARYGKVFRGNKVKNAQLAGAKGVILYSDPADYFAP
GVKSYPDGWNLPGGGVQRGNILNLNGAGDPLTPGYPANEYAYRRGIAEAVGLPSIPVH
PIGYYDAQKLLEKMGGSAPPDSSWRGSLKVPYNVGPGFTGNFSTQKVKMHIHSTNEVT
RIYNVIGTLRGAVEPDRYVILGGHRDSWVFGGIDPQSGAAVVHEIVRSFGTLKKEGWRP
RRTILFASWDAEEFGLLGSTEWAEENSRLLQERGVAYINADSSIEGNYTLRVDCTPLMY
SLVHNLTKELKSPDEGFEGKSLYESWTKKSPSPEFSGMPRISKLGSGNDFEVFFQRLGI
ASGRARYTKNWETNKFSGYPLYHSVYETYELVEKFYDPMFKYHLTVAQVRGGMVFELA
NSIVLPFDCRDYAVVLRKYADKIYSISMKHPQEMKTYSVSFDSLFSAVKNFTEIASKFSE
RLQDFDKSNPIVLRMMNDQLMFLERAFIDPLGLPDRPFYRHVIYAPSSHNKYAGESFPGI
YDALFDINNKVDTSKAWREVKRQISIAAFTVQAAAETLREVDSGDYKDDDDK 1047. Macaca
Macaca aa QDGNEEMGSITQTPYQVSISGTTILTC fascicularis fascicularis
CD3.epsilon. 1-27 1048. Macaca Macaca aa
QDGNEEMGSITQTPYQVSISGTTVILT fascicularis fascicularis CD3.epsilon.
1-27 1049. Macaca mulatta Macaca aa QDGNEEMGSITQTPYHVSISGTTVILT
CD3.epsilon. 1-27 mulatta
Sequence CWU 0 SQTB SEQUENCE LISTING The patent application
contains a lengthy "Sequence Listing" section. A copy of the
"Sequence Listing" is available in electronic form from the USPTO
web site
(http://seqdata.uspto.gov/?pageRequest=docDetail&DocID=US20190169310A1).
An electronic copy of the "Sequence Listing" will also be available
from the USPTO upon request and payment of the fee set forth in 37
CFR 1.19(b)(3).
0 SQTB SEQUENCE LISTING The patent application contains a lengthy
"Sequence Listing" section. A copy of the "Sequence Listing" is
available in electronic form from the USPTO web site
(http://seqdata.uspto.gov/?pageRequest=docDetail&DocID=US20190169310A1).
An electronic copy of the "Sequence Listing" will also be available
from the USPTO upon request and payment of the fee set forth in 37
CFR 1.19(b)(3).
* * * * *
References