U.S. patent application number 14/037841 was filed with the patent office on 2014-02-06 for lingo binding molecules and pharmaceutical use thereof.
This patent application is currently assigned to NOVARTIS AG. The applicant listed for this patent is Marta CORTES-CROS, Ingo Markus KLAGGE, Josef PRASSLER, Adrian Robert WALMSLEY, William Leonard WISHART. Invention is credited to Marta CORTES-CROS, Ingo Markus KLAGGE, Josef PRASSLER, Adrian Robert WALMSLEY, William Leonard WISHART.
Application Number | 20140037639 14/037841 |
Document ID | / |
Family ID | 38325542 |
Filed Date | 2014-02-06 |
United States Patent
Application |
20140037639 |
Kind Code |
A1 |
CORTES-CROS; Marta ; et
al. |
February 6, 2014 |
LINGO Binding Molecules and Pharmaceutical Use Thereof
Abstract
The present invention provides a binding molecule which is
capable of binding to the rat, cynomolgus monkey and human LINGO
polypeptide, a polynucleotide encoding such binding molecule; an
expression vector comprising said polynucleotide; an expression
system comprising a polynucleotide capable of producing a binding
molecule; an isolated host cell which comprises an expression
system as defined above; the use of such binding molecule as a
pharmaceutical, especially in the treatment to promote axonal
regeneration/plasticity; a pharmaceutical composition comprising
said binding molecule; and a method of treatment of diseases
associated with axonal degeneration and demyelination.
Inventors: |
CORTES-CROS; Marta; (Basel,
CH) ; KLAGGE; Ingo Markus; (Neuried, DE) ;
PRASSLER; Josef; (Germering, DE) ; WALMSLEY; Adrian
Robert; (Sierentz, FR) ; WISHART; William
Leonard; (Allschwil, CH) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
CORTES-CROS; Marta
KLAGGE; Ingo Markus
PRASSLER; Josef
WALMSLEY; Adrian Robert
WISHART; William Leonard |
Basel
Neuried
Germering
Sierentz
Allschwil |
|
CH
DE
DE
FR
CH |
|
|
Assignee: |
NOVARTIS AG
Basel
CH
|
Family ID: |
38325542 |
Appl. No.: |
14/037841 |
Filed: |
September 26, 2013 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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13616226 |
Sep 14, 2012 |
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14037841 |
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12514542 |
May 12, 2009 |
8299221 |
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PCT/EP07/09880 |
Nov 15, 2007 |
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13616226 |
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Current U.S.
Class: |
424/139.1 ;
435/320.1; 435/331; 530/387.3; 530/387.9; 536/23.53 |
Current CPC
Class: |
C07K 2317/77 20130101;
A61P 21/00 20180101; C07K 2317/55 20130101; A61P 25/28 20180101;
C07K 2317/75 20130101; C07K 16/18 20130101; C07K 2317/76 20130101;
A61P 25/00 20180101; C07K 2317/92 20130101; A61P 25/16 20180101;
C07K 2317/21 20130101; C07K 16/2803 20130101; C07K 2319/30
20130101; A61K 2039/505 20130101; A61P 27/02 20180101; A61P 43/00
20180101; C07K 16/28 20130101; C07K 2317/565 20130101; A61K
39/39533 20130101 |
Class at
Publication: |
424/139.1 ;
530/387.9; 530/387.3; 536/23.53; 435/320.1; 435/331 |
International
Class: |
C07K 16/18 20060101
C07K016/18 |
Foreign Application Data
Date |
Code |
Application Number |
Nov 17, 2006 |
EP |
06124350.7 |
Claims
1. A binding molecule which is capable of binding to the protein
according to SEQ ID NO: 1, SEQ ID NO: 2, or SEQ ID NO: 3, with a
dissociation constant <1000 nM.
2. A binding molecule according to claim 1 which binds to one or
more of the sequences chosen form the group consisting of SEQ ID
NO: 46-51.
3. A binding molecule according to claim 1 which is capable of
disinhibiting spinal cord myelin at a concentration of less than 20
nM.
4. A binding molecule according to claim 1 which is capable of
increasing the mean neurite length per cell of rat cerebellar
granule cells grown on a substrate of adult rat spinal cord myelin
by at least 20%.
5. A binding molecule according to claim 1, which comprises one or
more amino acid sequences chosen from the group consisting of SEQ
ID NO: 12-17 or SEQ ID NO: 18-23.
6. A binding molecule according to claim 5, which comprises at
least one antigen binding site chosen from the group consisting of;
a sequence which is at least 50% homologous to SEQ ID NO: 5 or SEQ
ID NO: 7, and; a sequence which is at least 50% homologous to SEQ
ID NO: 4 or SEQ ID NO: 6, or a direct equivalent thereof.
7. A binding molecule according to claim 6, which comprises a first
sequence which is at least 50% homologous to SEQ ID NO: 5 or SEQ ID
NO: 7, and a second sequence which is at least 50% homologous to
SEQ ID NO: 4 or SEQ ID NO: 6, or a direct equivalent thereof.
8. The binding molecule according to claim 5 which comprises at
least a) one immunoglobulin heavy chain or fragment thereof which
comprises (i) a variable domain comprising SEQ ID NO: 5 or SEQ ID
NO: 7, and (ii) the constant part or fragment thereof of a human
heavy chain; and b) one immunoglobulin light chain or fragment
thereof which comprises (i) a variable domain comprising SEQ ID NO:
4 or SEQ ID NO: 6, and (ii) the constant part or fragment thereof
of a human light chain; or direct equivalents thereof.
9. A binding molecule according to claim 5, which is an antibody or
a fragment thereof, or a direct equivalent thereof.
10. The binding molecule according to claim 9 in which the constant
part or fragment thereof of the human heavy chain is of the
.gamma.4 type and the constant part or fragment thereof of the
human light chain is of the .kappa. type.
11. The binding molecule according to claim 9, which is a human or
chimeric or humanized monoclonal antibody.
12. A polynucleotide encoding a binding molecule according to claim
9.
13. A polynucleotide chosen from the group consisting of SEQ ID NO:
8 and SEQ ID NO: 9; or from the group consisting of SEQ ID NO: 10
and SEQ ID NO: 11.
14. An expression vector comprising one or more polynucleotides
according to claim 12.
15. An expression system comprising a polynucleotide according to
claim 12, when said expression system or part thereof is present in
a compatible host cell.
16. An isolated host cell which comprises an expression system
according to claim 15.
17. A pharmaceutical composition comprising a binding molecule
according to claim 1 together with at least one pharmaceutically
acceptable carrier or diluent.
18. A method of treatment of diseases associated with the promotion
of axonal regeneration/plasticity comprising administering to a
subject in need of such treatment an effective amount of a binding
molecule according to claim 1.
Description
[0001] The present invention relates to LINGO binding molecules,
such as for example monoclonal antibodies or Fab fragments thereof,
and the use of such binding molecules for treating patients with
injuries to their central nervous system.
BACKGROUND OF THE INVENTION
[0002] Functional recovery following injury to the central nervous
system (CNS) of adult higher vertebrates is exceptionally limited,
resulting in persistent neurological deficits such as loss of limb
movement and sensation. As yet, there is a lack of an effective
therapy to treat humans with CNS injuries such as spinal cord
injury (SCI) and brain cortical injury. Although adult CNS neurons
generally survive axotomy, axonal regeneration is transitory and
only occurs over a confined area, hence retarding the re-formation
of functionally-relevant synaptic contacts. Furthermore, the
plastic capacity of the adult CNS is also restricted, thus
hindering the re-organisation of uninjured pathways to functionally
compensate for those ablated by the injury. Paradoxically,
axotomised axons in the peripheral nervous system (PNS) have a high
capacity to regenerate over long distances and frequently establish
functionally-meaningful connections (Schwab (2004) Curr Opin
Neurobiol 14, 118-124). This restriction in axonal
regeneration/plasticity is in part due to the expression on
myelinating oligodendrocytes of several proteins that have been
shown to be potent inhibitors of neurite outgrowth, namely Nogo-A
(Chen et al. (2000) Nature 403, 434-439; GrandPre et al. (2000)
Nature 403, 439-444; Prinjha et al. (2000) Nature 403, 383-384),
myelin-associated glycoprotein (MAG), and oligodendrocyte myelin
glycoprotein (OMgp) (McKerracher et al. (1994) Neuron 13, 805-811;
Wang et al. (2002) Nature 417:941-944) (FIG. 1A).
[0003] Nogo-A contains multiple neurite outgrowth inhibitory
domains exposed on the surface of oligodendrocytes: two are located
within the amino-terminal region (amino-Nogo-A) and one in the
C-terminal region (Nogo-66) (Oertle et al. (2003) J Neurosci 23,
5393-5406). Nogo-66 binds and signals through a
glycosyl-phosphatidylinositol (GPI)-anchored leucine-rich repeat
(LRR)-containing receptor on the neuronal surface known as the
Nogo-66 receptor (NgR) (Fournier et al. (2001) Nature 409,
341-346). Although structurally unrelated, MAG and OMgp also bind
and signal through NgR (Domeniconi et al. (2002) Neuron 35,
283-290; Liu et al. (2002) Science 297, 1190-1193; Wang et al.
(2002) Nature 417:941-944). Signaling through NgR leads to the
activation of the small GTPase RhoA which in turn activates
Rho-associated kinase (ROCK) leading to a rigidification of the
actin cytoskeleton and inhibition of axonal extension (Niederost et
al. (2002) J Neurosci 22, 10368-10376; Schweigreiter et al. (2004)
Mol Cell Neurosci 27:163-174). All three ligands bind within the
LRR region of NgR and have partially over-lapping binding sites
(Fournier et al. (2002) J Neurosci 22, 8876-8883; Liu et al. (2002)
Science 297, 1190-1193; Wang et al. (2002) Nature 417:941-944;
Barton et al. (2003) EMBO J 22, 3291-3302). The receptor(s) for the
inhibitory domains within amino-Nogo-A are unknown but have been
shown to be distinct from NgR (Schweigreiter et al. (2004) Mol Cell
Neurosci 27:163-174). MAG has also been found to signal through a
close homologue of NgR known as NgR2 (Pignot et al. (2003) J
Neurochem 85, 717-728; Venkatesh et al. (2005) J Neurosci 25,
808-822).
[0004] As NgR lacks a cytoplasmic domain, it utilizes several
transmembrane proteins for signal transduction, namely the low
affinity neurotrophin receptor p75.sup.NTR, TROY (a.k.a. TAJ) and
LINGO-1 (LRR and Ig domain-containing, Nogo receptor-interacting
protein a.k.a LRRN6A or LERN1) (Wang et al. (2002) Nature 420,
74-78; Carim-Todd et al. (2003) Eur J Neurosci 18, 3167-3182; Mi et
al. (2004) Nat Neurosci 7, 221-228; Park et al. (2005) Neuron
45:345-351; Shao et al. (2005) Neuron 45, 353-359). TROY and
p75.sup.NTR can functionally replace each other in the NgR receptor
complex, whereas the presence of LINGO-1 is an absolute
prerequisite for signaling to occur. The NgR receptor complex is
therefore seen as a ternary complex comprising NgR as the ligand
binding subunit and LINGO-1 as the common signal transducing
subunit acting in concert with either p75.sup.NTR or TROY.
[0005] LINGO-1 is a single transmembrane protein expressed
exclusively within the CNS predominantly on neurons and
oligodendrocytes. The expression of LINGO-1 peaks in the early
postnatal period and is up-regulated in the adult spinal cord upon
injury. The ectodomain of LINGO-1 contains twelve tandem LRRs
flanked by N- and C-terminal subdomains followed by a basic region
and an Ig domain (FIG. 1B). Given that an AP fusion of the LINGO-1
ectodomain bound to COS-7 cells expressing NgR or p75.sup.NTR or
both and, similarly, LINGO-1 co-precipitated with NgR or
p75.sup.NTR in cells expressing all three proteins, NTR LINGO-1
most likely forms a ternary complex with NgR and p75.sup.NTR by
interacting with both simultaneously.
[0006] In addition to being expressed on neurons, LINGO-1 is also
expressed in oligodendrocytes in the adult CNS (Mi et al. (2005)
Nat Neurosci 8, 745-751). Inhibiting LINGO-1 signaling in
oligodendrocyte cultures by either treatment with LINGO-1-Fc,
down-regulation of the protein with RNAi or over-expression of
DN-LINGO-1 augmented the differentiation of OPCs to myelinating
oligodendrocytes. Furthermore, genetic ablation of LINGO-1 in mice
increased the number of mature oligodendrocytes and,
correspondingly, myelinated axons in the spinal cord. Inhibition of
LINGO-1 signaling reduced the activation of RhoA and increased the
activity of Fyn kinase, both of which are reported to promote
oligodendrocyte differentiation, although the actual
ligands/interactions responsible for activating LINGO-1 signaling
have yet to be exemplified. This has led to the conclusion the
LINGO-1 is a negative regulator of myelination.
[0007] Multiple Sclerosis (MS) is a chronic inflammatory disease of
the CNS characterised by demyelination and axonal degeneration
leading to multiple neurological deficits. Although remyelination
of axons can occur early in the disease, at some point
remyelination fails completely leading to accelerated axonal
degeneration and irreversible damage. Remyelination most likely
arises from the differentiation of adult oligodendrocyte precursor
cells (OPCs) which migrate to the margins of active lesions. As
LINGO-1 negatively regulates myelination, blockade of LINGO-1 may
augment remyelination, attenuate axonal degeneration, promote
axonal regeneration and thus attenuate, halt or even reverse the
progress of demyelinating diseases such as MS.
[0008] Blockade of LINGO-1 has also been shown to improve the
survival of dopaminergic neurons and reduce behavioural
abnormalities in rodent models of Parkinson's disease (Inoue et al.
(2007) Proc Natl Acad Sci USA 104, 14430-14435).
SUMMARY OF THE INVENTION
[0009] It has now surprisingly been found that novel monoclonal
human antibodies against LINGO-1 (known as antibody 4784, and
antibody 4785 hereafter) significantly inhibit the association of
LINGO-1 with NgR and significantly attenuate the neurite outgrowth
inhibitory activity of adult rat spinal cord myelin at sub-nM
concentrations in vitro. In addition, the said antibodies
significantly increase the differentiation of primary
oligodendrocytes in vitro and have been shown to significantly
downregulate cell surface LINGO-1 in living cells. Treatment with
these antibodies is expected to increase axonal
regeneration/plasticity and improve functional recovery following
acute CNS injuries such as SCI and brain cortical injury.
Furthermore, blocking LINGO-1 signaling using the said antibodies
in oligodendroglial cells has the potential to augment the
remyelination of axons in demyelinating diseases such as MS leading
to an attenuation of disease progression. In concert, inhibiting
LINGO-1 signaling in neurons with the said antibodies can be
expected to improve axonal regeneration and neuroplasticity and
promote the recovery of neurological function lost during the
course of the disease. Finally, blockade of LINGO-1 with the said
antibodies can be expected to attenuate the pathogenesis of
Parkinson's disease.
[0010] Furthermore, the invention provides binding molecules which
bind to specific epitopes on LINGO-1.
[0011] The antibodies have sub-nM K.sub.Ds against the rat,
cynomolgus monkey and human LINGO-1 ectodomain, significantly
attenuate the neurite outgrowth inhibitory activity of adult rat
spinal cord myelin at sub-nM concentrations and significantly
increase oligodendrocyte differentiation in vitro. Moreover, it is
now possible to construct other LINGO-1 binding molecules having
the same variable regions as said antibodies.
DETAILED DESCRIPTION OF THE INVENTION
[0012] Accordingly, the invention provides binding molecules to a
particular region or epitope of LINGO-1 (hereinafter referred to as
"the binding molecules of the invention" or simply "binding
molecules").
[0013] The binding molecules of the invention bind the mature
ectodomain (residues 34-550) of rat LINGO-1 (SEQ ID NO: 1),
cynomolgus monkey LINGO-1 (SEQ ID NO: 2) and human LINGO-1 (SEQ ID
NO: 3) with a dissociation constant (K.sub.D)<1000 nM, more
preferably with a K.sub.D<100 nM, most preferably with a
K.sub.D<10 nM. The binding reaction may be shown by standard
methods (qualitative assays) including, for example, the FACS
method described in Examples. In addition, the binding to rat,
cynomolgus monkey and human LINGO-1, and also the efficiency, may
be shown in a neurite outgrowth assay and oligodendrocyte assay as
described below.
[0014] Thus, in a further preferred embodiment the binding
molecules (at a concentration of 100 nM, preferably 10 nM, more
preferably at 1 nM even more preferably at 0.1 nM) increase the
mean neurite length per cell of rat cerebellar granule cells grown
on a substrate of adult rat spinal cord myelin by at least 20%,
preferably 50%, most preferred 60% compared to the mean neurite
length per cell of rat cerebellar granule cells which are treated
with a control antibody that does not bind to the rat, cynomolgous
monkey and human LINGO-1 ectodomain.
[0015] By using peptide microarrays, the specific epitope to which
the binding molecules of the invention bind is determined according
to methods well known in the art. Consequently, in another
embodiment the invention provides binding molecules which bind to
at least one of the LINGO-1 epitopes as defined by SEQ ID NO:
46-51. SEQ ID NO: 46: KIVILLDYMFQD, SEQ ID NO: 47: AIRDYSFKRLYR,
SEQ ID NO: 48: LKVLEISHWPYL, SEQ ID NO: 49: NLTAVPYLAVRHLVY, SEQ ID
NO: 50: YFTCRRARI, or SEQ ID NO: 51: DVLLPNYFTCRRARI.
[0016] In another embodiment, the binding molecules of the
invention comprises one or more, of the following CDR sequences,
e.g. all of the Antibody 4784 or all of the Antibody 4785 sequences
mentioned there:
TABLE-US-00001 (Antibody 4784 CDR-H1) SEQ ID NO: 12 SSGVGVG
(Antibody 4784 CDR-H2) SEQ ID NO: 13 HIGSDDDKYYSTSLKT (Antibody
4784 CDR-H3) SEQ ID NO: 14 NQQYGDGYPGYFDY (Antibody 4784 CDR-L1)
SEQ ID NO: 15 SGDNIGNYYVY (Antibody 4784 CDR-L2) SEQ ID NO: 16
EDTNRPS (Antibody 4784 CDR-L3) SEQ ID NO: 17 QSYDNLHEQV (Antibody
4785 CDR'-H1) SEQ ID NO: 18 DNSAAWS (Antibody 4785 CDR'-H2) SEQ ID
NO: 19 LIYLRSKWDNDYAVSVKS (Antibody 4785 CDR'-H3) SEQ ID NO: 20
TGRADEFDV (Antibody 4785 CDR'-L1) SEQ ID NO: 21 SGSSSNIGNNYVS
(Antibody 4785 CDR'-L2) SEQ ID NO: 22 RNSKRPS (Antibody 4785
CDR'-L3) SEQ ID NO: 23 STYDTFSIV
[0017] More preferably, the binding molecules comprise one or more
of the sequences given above for Antibody 4784 with the SEQ ID NO:
12, 13, 14, 15, 16 and/or 17; or for Antibody 4785 with the SEQ ID
NO: 18, 19, 20, 21, 22 and/or 23.
[0018] Those skilled in the art understand that changes can be made
to 4784 or 4785 which, though they change several, more preferably
one or more amino acids, preferably up to three, e.g. one or two,
of the SDRs given above, especially in one or more or all of them,
e.g. one or two of them, or provide alternative post-translational
modification of product formats, result in a therapeutic agent
demonstrating the same or substantially similar anti-Lingo-1
binding behaviour.
[0019] In another embodiment the binding molecules of the invention
comprises at least one antigen binding site chosen from the group
consisting of; a sequence which is at least 50%, at least 60%, at
least 70%, at least 80%, at least 90%, at least 95%, at least 97%,
or at least 99% homologous to SEQ ID NO: 5 or SEQ ID NO: 7,
and;
a sequence which is at least 50%, at least 60%, at least 70%, at
least 80%, at least 90%, at least 95%, at least 97%, or at least
99% homologous to SEQ ID NO: 4 or SEQ ID NO: 6, or a direct
equivalent thereof.
[0020] In one embodiment, the binding molecule comprises at least
one binding site chosen from the group consisting of SEQ ID NO: 5
or SEQ ID NO: 7, and;
SEQ ID NO: 4 or SEQ ID NO: 6.
[0021] The invention further provides a binding molecule which
comprises a first sequence which is at least 50%, at least 60%, at
least 70%, at least 80%, at least 90%, at least 95%, at least 97%,
or at least 99% homologous to SEQ ID NO: 5, and a second sequence
which is at least 50% at least 60%, at least 70%, at least 80%, at
least 90%, at least 95%, at least 97%, or at least 99% homologous
to SEQ ID NO: 4, or a direct equivalent thereof.
[0022] The invention further provides a binding molecule which
comprises a first sequence which is at least 50%, at least 60%, at
least 70%, at least 80%, at least 90%, at least 95%, at least 97%,
or at least 99% homologous to SEQ ID NO: 7, and a second sequence
which is at least 50% at least 60%, at least 70%, at least 80%, at
least 90%, at least 95%, at least 97%, or at least 99% homologous
to SEQ ID NO: 6, or a direct equivalent thereof.
[0023] In one embodiment, the invention provides a binding molecule
according to claims 1 to 7 which comprises at least [0024] a) one
immunoglobulin heavy chain or fragment thereof which comprises
[0025] (i) a variable domain comprising SEQ ID NO: 5 or SEQ ID NO:
7, and [0026] (ii) the constant part or fragment thereof of a human
heavy chain; and [0027] b) one immunoglobulin light chain or
fragment thereof which comprises [0028] (i) a variable domain
comprising SEQ ID NO: 4 or SEQ ID NO: 6, and [0029] (ii) the
constant part or fragment thereof of a human light chain; or [0030]
direct equivalents thereof; e.g. two or three of each of the chains
given under a) or b).
[0031] The sequences may be at least 50%, at least 60%, at least
70%, at least 80%, at least 90%, at least 95%, at least 97%, or at
least 99% homologous to SEQ ID NO: 4-7. The important factor is
that such variants retain the binding capabilities to LINGO-1, the
disinhibitory effect (especially the ability to attenuate the
neurite outgrowth inhibitory activity of adult rat spinal cord
myelin at sub-nM concentrations), and/or to improve the functional
recovery of SCI (especially in a rat model), in each case
preferably as described in the Examples or the remaining
description.
[0032] In one embodiment, the invention provides a binding molecule
which is an antibody comprising one or more of the sequences
according to SEQ ID NO: 4-7 or SEQ ID NO: 12-23, or a fragment
thereof, or a direct equivalent thereof.
[0033] In a further embodiment, the binding molecule, as an
antibody, has a constant part or fragment thereof of the human
heavy chain of the .gamma.4 type and the constant part or fragment
thereof of the human light chain is of the .lamda. type.
[0034] In a further embodiment, the binding molecule, as an
antibody, has a constant part or fragment thereof of the human
heavy chain of the .gamma.4 type and the constant part or fragment
thereof of the human light chain is of the .kappa. type.
[0035] In a further embodiment, the binding molecule is a human or
chimeric or humanized monoclonal antibody.
[0036] In a further embodiment, the binding molecule is a
humaneered antibody.
[0037] The invention also provides a polynucleotide encoding a
binding molecule as defined above.
[0038] The polynucleotide may be chosen from the group consisting
of SEQ ID NO: 8 and SEQ ID NO: 9; or from the group consisting of
SEQ ID NO: 10 and SEQ ID NO: 11.
[0039] The invention also provides an expression vector comprising
one or more polynucleotides according to SEQ ID NO:8-11.
[0040] Furthermore, the invention provides an expression system
comprising a polynucleotide according to SEQ ID NO:8-11, wherein
said expression system or part thereof is capable of producing a
binding molecule as set out above, when said expression system or
part thereof is present in a compatible host cell. The invention
also provides an isolated host cell which comprises such an
expression system.
[0041] The invention also provides the use of a binding molecule as
set out above, as a medicament.
[0042] The invention also provides the use of a binding molecule as
set out above in the preparation of a medicament for the treatment
of a CNS injury.
[0043] The invention also provides a pharmaceutical composition
comprising a binding molecule as set out above together with at
least one pharmaceutically acceptable carrier or diluent.
[0044] Furthermore, the invention provides a method of treatment of
diseases associated with the promotion of axonal
regeneration/plasticity comprising administering to a subject in
need of such treatment an effective amount of a binding molecule as
set out above.
[0045] The invention also provides a method of treatment of
diseases associated with the promotion of axonal
regeneration/plasticity comprising administering to a subject in
need of such treatment an effective amount of a binding molecule
according to any one of claims 1 to 10.
[0046] When the antigen binding site comprises both the first and
second domains, these may be located on the same polypeptide
molecule or, preferably, each domain may be on a different chain,
the first domain being part of an immunoglobulin heavy chain or
fragment thereof and the second domain being part of an
immunoglobulin light chain or fragment thereof.
[0047] Examples of binding molecules of the invention include
antibodies as produced by phage display and human or chimeric
humanized antibodies, or further humaneered antibodies, or any
fragment thereof, e.g. F(ab')2; and Fab fragments, as well as
single chain or single domain antibodies. The term "antibody" is
meant to include such binding molecules.
[0048] A single chain antibody consists of the variable domains of
an antibody heavy and light chains covalently bound by a peptide
linker usually consisting of from 10 to 30 amino acids, preferably
from 15 to 25 amino acids. Therefore, such a structure does not
include the constant part of the heavy and light chains and it is
believed that the small peptide spacer should be less antigenic
than a whole constant part. By "chimeric antibody" is meant an
antibody in which the constant regions of heavy or light chains or
both are of human origin while the variable domains of both heavy
and light chains are of non-human (e.g. murine) origin. By
"humanized antibody" is meant an antibody in which the
hypervariable regions (CDRs) are of non-human (e.g. murine) origin,
while all or substantially all the other parts of the
immunoglobulin e.g. the constant regions and the highly conserved
parts of the variable domains, i.e. the framework regions, are of
human origin. A humanized antibody may however retain a few amino
acids of the murine sequence in the parts of the framework regions
adjacent to the hypervariable regions.
[0049] Hypervariable regions may be associated with any kind of
framework regions, preferably of murine or human origin. Suitable
framework regions are described in "Sequences of proteins of
immunological interest" (Kabat E. A. et al, US department of health
and human services, Public health service, National Institute of
Health, preferably incorporated herein, especially with regard to
the framework regions, by reference). Preferably the constant part
of a human heavy chain of the binding molecules may be of the
.gamma.4 type, including subtypes, preferably the constant part of
a human light chain may be of the .kappa. or .lamda. type, more
preferably of the .lamda. type.
[0050] A naturally occurring "antibody" is a glycoprotein
comprising at least two heavy (H) chains and two light (L) chains
inter-connected by disulfide bonds. Each heavy chain is comprised
of a heavy chain variable region (abbreviated herein as V.sub.H)
and a heavy chain constant region. The heavy chain constant region
is comprised of three domains, CH1, CH2 and CH3. Each light chain
is comprised of a light chain variable region (abbreviated herein
as V.sub.L) and a light chain constant region. The light chain
constant region is comprised of one domain, C.sub.L. The V.sub.H
and V.sub.L regions can be further subdivided into regions of
hypervariability, termed complementarity determining regions (CDR),
interspersed with regions that are more conserved, termed framework
regions (FR). Each V.sub.H and V.sub.L is composed of three CDRs
and four FRs arranged from amino-terminus to carboxy-terminus in
the following order: FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4. The
variable regions of the heavy and light chains contain a binding
domain that interacts with an antigen. The constant regions of the
antibodies may mediate the binding of the immunoglobulin to host
tissues or factors, including various cells of the immune system
(e.g., effector cells) and the first component (C1q) of the
classical complement system.
[0051] The term "antigen-binding portion" of an antibody (or simply
"antigen portion"), as used herein, refers to full length or one or
more fragments of an antibody that retain the ability to
specifically bind to an antigen (e.g., LINGO-1 and/or LINGO-2). It
has been shown that the antigen-binding function of an antibody can
be performed by fragments of a full-length antibody. Examples of
binding fragments encompassed within the term "antigen-binding
portion" of an antibody include a Fab fragment, a monovalent
fragment consisting of the V.sub.L, V.sub.H, C.sub.L and CH1
domains; a F(ab').sub.2 fragment, a bivalent fragment comprising
two Fab fragments linked by a disulfide bridge at the hinge region;
a Fd fragment consisting of the V.sub.H and CH1 domains; a Fv
fragment consisting of the V.sub.L and V.sub.H domains of a single
arm of an antibody; a dAb fragment (Ward et al., 1989 Nature
341:544-546), which consists of a V.sub.H domain; and an isolated
complementarity determining region (CDR).
[0052] The terms "monoclonal antibody" or "monoclonal antibody
composition" as used herein refer to a preparation of antibody
molecules of single molecular composition (that is, that are
identical because they are produced by one type of immune cell that
are all clones of a single parent cell). A monoclonal antibody
composition displays an (essentially) single binding specificity
and affinity for a particular epitope.
[0053] The term "human antibody", as used herein, is intended to
include antibodies having variable regions in which both the
framework and CDR regions are derived from sequences of human
origin. Furthermore, if the antibody contains a constant region,
the constant region also is derived from such human sequences,
e.g., human germline sequences, or mutated versions of human
germline sequences. The human antibodies of the invention may
include amino acid residues not encoded by human sequences (e.g.,
mutations introduced by random or site-specific mutagenesis in
vitro or by somatic mutation in vivo). However, the term "human
antibody", as used herein, is not intended to include antibodies in
which CDR sequences derived from the germline of another mammalian
species, such as a mouse, have been grafted onto human framework
sequences.
[0054] The term "human monoclonal antibody" refers to antibodies
displaying an (essentially) single binding specificity which have
variable regions in which both the framework and CDR regions are
derived from human sequences. In one embodiment, the human
monoclonal antibodies are produced by a hybridoma which includes a
B cell obtained from a transgenic nonhuman animal, e.g., a
transgenic mouse, having a genome comprising a human heavy chain
transgene and a light chain transgene fused to an immortalized
cell.
[0055] The term "recombinant human antibody", as used herein,
includes all human antibodies that are prepared, expressed, created
or isolated by recombinant means, such as antibodies isolated from
an animal (e.g., a mouse) that is transgenic or transchromosomal
for human immunoglobulin genes or a hybridoma prepared therefrom,
antibodies isolated from a host cell transformed to express the
human antibody, e.g., from a transfectoma, antibodies isolated from
a recombinant, combinatorial human antibody library, and antibodies
prepared, expressed, created or isolated by any other means that
involve splicing of all or a portion of a human immunoglobulin
gene, sequences to other DNA sequences. Such recombinant human
antibodies have variable regions in which the framework and CDR
regions are derived from human germline immunoglobulin sequences.
In certain embodiments, however, such recombinant human antibodies
can be subjected to in vitro mutagenesis (or, when an animal
transgenic for human Ig sequences is used, in vivo somatic
mutagenesis) and thus the amino acid sequences of the V.sub.H and
V.sub.L regions of the recombinant antibodies are sequences that,
while derived from and related to human germline V.sub.H and
V.sub.L sequences, may not naturally exist within the human
antibody germline repertoire in vivo.
[0056] As used herein, "isotype" refers to the antibody class
(e.g., IgM, IgE, IgG such as IgG1 or IgG4) that is provided by the
heavy chain constant region genes.
[0057] As used herein, the term "Affinity" refers to the strength
of interaction between antibody and antigen at single antigenic
sites. Within each antigenic site, the variable region of the
antibody "arm" interacts through weak non-covalent forces with
antigen at numerous sites; the more interactions, the stronger the
affinity.
[0058] The term "K.sub.D", as used herein, is intended to refer to
the dissociation constant, which is obtained from the ratio of
K.sub.d to K.sub.a (association rate to dissociation rate) (i.e.
K.sub.d/K.sub.a) and is expressed as a molar concentration (M).
K.sub.D values for antibodies can be determined using methods well
established in the art. A method for determining the K.sub.D of an
antibody is by using surface plasmon resonance, or using a
biosensor system such as a Biacore.RTM. system.
[0059] A binding molecule according to the invention is preferably
an "isolated antibody", which, as used herein, refers to an
antibody that is substantially free of other antibodies having
different antigenic specificities (e.g., an isolated antibody that
specifically binds LINGO-1, LINGO-2 or LINGO-1 and LINGO-2 is
substantially free of antibodies that specifically bind antigens
other than those mentioned). An isolated antibody that specifically
binds may, however, have cross-reactivity to other antigens, such
as LINGO-1 or LINGO-2 molecules from other species. Moreover, an
isolated antibody is preferably substantially free of other
cellular material and/or chemicals.
[0060] The invention also provides a binding molecule of the
invention which may be selected from a single chain binding
molecule which comprises an antigen binding site (especially with
the CDRs described above for Antibody 4784) of antibody 4784
comprising [0061] a) a first domain comprising the variable
sequence of the heavy chain having the amino acid sequence (SEQ ID
NO: 5) [0062] b) a second domain comprising the variable sequence
of the light chain having the amino acid sequence (SEQ ID NO: 4)
[0063] c) a peptide linker which is bound either to the N-terminal
extremity of the first domain and to the C-terminal extremity of
the second domain or to the C-terminal extremity of the first
domain and to the N-terminal extremity of second domain; or direct
equivalents thereof.
[0064] A binding molecule of the invention may be selected from a
single chain binding molecule which comprises an antigen binding
site (especially with the CDRs described above for Antibody 4785)
of antibody 4785 comprising [0065] a) a first domain comprising the
variable sequence of the heavy chain having the amino acid sequence
(SEQ ID NO: 7) [0066] b) a second domain comprising the variable
sequence of the light chain having the amino acid sequence (SEQ ID
NO: 6) [0067] c) a peptide linker which is bound either to the
N-terminal extremity of the first domain and to the C-terminal
extremity of the second domain or to the C-terminal extremity of
the first domain and to the N-terminal extremity of second domain;
or direct equivalents thereof.
[0068] As it is well known, minor changes in an amino acid sequence
such as deletion, addition or substitution of one or several amino
acids may lead to an allelic form of the original protein which has
substantially identical properties. Thus, by the term "direct
equivalents thereof" is meant either any single domain binding
molecule of the invention (molecule X) [0069] (i) in which the
variable region of the binding molecule (e.g. SEQ ID NO: 4, 5, 6 or
7) is at least 50 or 80% homologous, preferably at least 90%
homologous, more preferably at least 95, 96, 97, 98, 99% homologous
to the equivalent variable regions of the light and heavy chains
comprising the direct equivalents of SEQ ID NO: 4 and SEQ ID NO: 5,
respectively or light and heavy chains comprising the direct
equivalents of SEQ ID NO: 6 and SEQ ID NO: 7, respectively). [0070]
(ii) which is capable of binding to the ectodomain (residues
34-550) of rat LINGO-1 (SEQ ID NO: 1), cynomolgus monkey LINGO-1
(SEQ ID NO: 2) and human LINGO-1 (SEQ ID NO: 3), preferably with a
dissociation constant (K.sub.D)<1000 nM, more preferably with a
K.sub.D<100 nM, most preferably with a K.sub.D<10 nM, or any
binding molecule of the invention having at least two domains per
binding site (molecule X').
[0071] Thus further embodiments of the inventions are for example a
binding molecule which is capable of binding to the ectodomain of
rat, cynomolgus monkey and/or human LINGO-1 with a dissociation
constant <1000 nM and comprises at least one antigen binding
site, said antigen binding site comprising in sequence the variable
region which is at least 50%, preferably 80, 90, 95, 96, 97, 98,
99% homologous to the equivalent variable regions of the light and
heavy chains of 4784 (SEQ ID NO: 4 and SEQ ID NO: 5, respectively)
or light and heavy chains of 4785 (SEQ ID NO: 6 and SEQ ID NO: 7,
respectively).
[0072] In another embodiment, the binding molecule comprises at
least one amino acid sequence chosen from the group consisting of
SEQ ID NO: 12-23, or a sequence which is at least 50%, preferably
80, 90, 95, 96, 97, 98, 99% homologous to these sequences.
[0073] This dissociation constant may be conveniently tested in
various assays including, for example, the FACS method described in
the examples. In addition, the binding and functional effect of the
binding molecules may be shown in a bioassay, e.g. the neurite
outgrowth assay as described below.
[0074] The constant part of a human heavy chain may be of the
.gamma.1; .gamma.2; .gamma.3; .gamma.4; .alpha.1; .alpha.2; .delta.
or .epsilon. type, preferably of the .gamma. type, more preferably
of the .gamma.4 type, whereas the constant part of a human light
chain may be of the .kappa. or .lamda. type (which includes the
.lamda.1; .lamda.2; and .lamda.3 subtypes) but is preferably of the
.lamda. type. The amino acid sequence of all these constant parts
are given in Kabat et al (Supra).
[0075] Conjugates of the binding molecules of the invention, e.g.
enzyme or toxin or radioisotope conjugates, are also included
within the scope of the invention.
[0076] "Polypeptide", if not otherwise specified herein, includes
any peptide or protein comprising amino acids joined to each other
by peptide bonds, having an amino acid sequence starting at the
N-terminal extremity and ending at the C-terminal extremity.
Preferably, the polypeptide of the present invention is a
monoclonal antibody, more preferred is a chimeric (also called
V-grafted) or humanised (also called CDR-grafted) monoclonal
antibody. The humanised (CDR-grafted) monoclonal antibody may or
may not include further mutations introduced into the framework
(FR) sequences of the acceptor antibody.
[0077] A functional derivative of a polypeptide as used herein
includes a molecule having a qualitative biological activity in
common with a polypeptide to the present invention, i.e. having the
ability to bind to the ectodomain of rat, cynomolgus monkey and
human LINGO-1.
[0078] A functional derivative includes fragments and peptide
analogs of a polypeptide according to the present invention. It
also includes the term "direct derivatives".
[0079] Fragments comprise regions within the sequence of a
polypeptide according to the present invention, e.g. of a specified
sequence. Fragments of binding molecules, especially of antibodies,
are functional fragments, i.e. they comprise at least one portion
capable of binding to LINGO-1 and/or LINGO-2, especially to at
least one of the epitopes given by SEQ ID NO: 46, 47, 48, 49, 50
and 51, preferably with the binding affinities (K.sub.D) mentioned
above or in the Examples, especially as being preferred.
[0080] The term "derivative" is used to define amino acid sequence
variants, and covalent modifications of a polypeptide according to
the present invention. e.g. of a specified sequence. The functional
derivatives of a polypeptide according to the present invention,
e.g. of a specified sequence, e.g. of the hypervariable region of
the light and the heavy chain, preferably have at least about 65%,
more preferably at least about 75%, even more preferably at least
about 85%, most preferably at least about 95, 96, 97, 98, 99%
overall sequence homology with the amino acid sequence of a
polypeptide according to the present invention, e.g. of a specified
sequence, and substantially retain the ability to bind the
ectodomain of rat, cynomolgus monkey and human LINGO-1 (and
optionally in addition to LINGO-2).
[0081] The term "covalent modification" includes modifications of a
polypeptide according to the present invention, e.g. of a specified
sequence; or a fragment thereof with an organic proteinaceous or
non-proteinaceous derivatizing agent, fusions to heterologous
polypeptide sequences, and post-translational modifications.
Covalent modified polypeptides, e.g. of a specified sequence, still
have the ability to bind to the ectodomain of rat, cynomolgus
monkey and human LINGO-1. Covalent modifications are traditionally
introduced by reacting targeted amino acid residues with an organic
derivatizing agent that is capable of reacting with selected sides
or terminal residues, or by harnessing mechanisms of
post-translational modifications that function in selected
recombinant host cells. Certain post-translational modifications
are the result of the action of recombinant host cells on the
expressed polypeptide. Glutaminyl and asparaginyl residues are
frequently post-translationally deaminated to the corresponding
glutamyl and aspartyl residues. Alternatively, these residues are
deaminated under mildly acidic conditions. Other post-translational
modifications include hydroxylation of proline and lysine,
phosphorylation of hydroxyl groups of seryl, tyrosine or threonyl
residues, methylation of the .alpha.-amino groups of lysine,
arginine, and histidine side chains, see e.g. T. E. Creighton,
Proteins: Structure and Molecular Properties, W. H. Freeman &
Co., San Francisco, pp. 79-86 (1983). Covalent modifications e.g.
include fusion proteins comprising a polypeptide according to the
present invention, e.g. of a specified sequence and their amino
acid sequence variants, such as immunoadhesins, and N-terminal
fusions to heterologous signal sequences.
[0082] "Homology" (or "identity) with respect to a native
polypeptide and its functional derivative is defined herein as the
percentage of amino acid residues in the candidate sequence that
are identical with the residues of a corresponding native
polypeptide, after aligning the sequences and introducing gaps, if
necessary, to achieve the maximum percent homology, and not
considering any conservative substitutions as part of the sequence
identity. Neither N- or C-terminal extensions nor insertions shall
be construed as reducing identity or homology. Methods and computer
programs for the alignment are well known.
[0083] Preferably, as used herein, the percent homology between two
amino acid sequences or two nucleotide sequences is equivalent to
the percent identity between the two sequences. The percent
identity between the two sequences is a function of the number of
identical positions shared by the sequences (i.e., % homology=# of
identical positions/total # of positions .times.100), taking into
account the number of gaps, and the length of each gap, which need
to be introduced for optimal alignment of the two sequences. The
comparison of sequences and determination of percent identity
between two sequences can be accomplished using a mathematical
algorithm, as described in the non-limiting examples below:
[0084] The percent identity between two amino acid sequences can be
determined using the algorithm of E. Meyers and W. Miller (Comput.
Appl. Biosci., 4:11-17, 1988) which has been incorporated into the
ALIGN program (version 2.0), using a PAM120 weight residue table, a
gap length penalty of 12 and a gap penalty of 4. In addition, the
percent identity between two amino acid sequences can be determined
using the Needleman and Wunsch (J. Mol, Biol. 48:444-453, 1970)
algorithm which has been incorporated into the GAP program in the
GCG software package (available at http://www.gcg.com), using
either a Blossom 62 matrix or a PAM250 matrix, and a gap weight of
16, 14, 12, 10, 8, 6, or 4 and a length weight of 1, 2, 3, 4, 5, or
6.
[0085] Additionally or alternatively, the protein sequences of the
present invention can further be used as a "query sequence" to
perform a search against public databases to, for example, identify
related sequences. Such searches can be performed using the XBLAST
program (version 2.0) of Altschul, et al., 1990 J. Mol. Biol.
215:403-10. BLAST protein searches can be performed with the XBLAST
program, score=50, wordlength=3 to obtain amino acid sequences
homologous to the antibody molecules of the invention. To obtain
gapped alignments for comparison purposes, Gapped BLAST can be
utilized as described in Altschul et al., 1997 Nucleic Acids Res.
25(17):3389-3402. When utilizing BLAST and Gapped BLAST programs,
the default parameters of the respective programs (e.g., XBLAST and
NBLAST) can be used. See http:www.ncbi.nhn.nih.gov.
[0086] "Amino acid(s)" refer to all naturally occurring
L-.alpha.-amino acids, e.g. and including D-amino acids. The amino
acids are identified by either the well known single-letter or
three-letter designations.
[0087] The term "amino acid sequence variant" refers to molecules
with some differences in their amino acid sequences as compared to
a polypeptide according to the present invention, e.g. of a
specified sequence. Amino acid sequence variants of a polypeptide
according to the present invention, e.g. of a specified sequence,
still have the ability to bind to the ectodomain of rat, cynomolgus
monkey and human LINGO-1. Substitutional variants are those that
have at least one amino acid residue removed and a different amino
acid inserted in its place at the same position in a polypeptide
according to the present invention, e.g. of a specified sequence.
These substitutions may be single, where only one amino acid in the
molecule has been substituted, or they may be multiple, where two
or more, e.g. 1 to 10, preferably 1 to 5, more preferably 1 to 3,
amino acids have been substituted in the same molecule. Insertional
variants are those with one or more, e.g. 1 to 100, such as 1 to
10, amino acids inserted immediately adjacent to an amino acid at a
particular position in a polypeptide according to the present
invention, e.g. of a specified sequence. Immediately adjacent to an
amino acid means connected to either the .alpha.-carboxy or
.alpha.-amino functional group of the amino acid. Deletional
variants are those with one or more, e.g. 1 to 100, such as 1 to 10
or 1 to 5, amino acids in a polypeptide according to the present
invention, e.g. of a specified sequence, removed. Ordinarily,
deletional variants will have one or two amino acids deleted in a
particular region of the molecule.
[0088] A binding molecule of the invention may be produced by
recombinant DNA techniques. In view of this, one or more DNA
molecules encoding the binding molecule must be constructed, placed
under appropriate control sequences and transferred into a suitable
host organism for expression.
[0089] In a very general manner, there are accordingly provided
[0090] (i) DNA molecules encoding a single domain binding molecule
of the invention, a single chain binding molecule of the invention,
a heavy or light chain or fragments thereof of a binding molecule
of the invention; and [0091] (ii) the use of the DNA molecules of
the invention for the production of a binding molecule of the
invention by recombinant means.
[0092] The present state of the art is such that the skilled person
will be able to synthesize the DNA molecules of the invention given
the information provided herein i.e. the amino acid sequences of
the hypervariable regions and the DNA sequences coding for them. A
method for constructing a variable domain gene is for example
described in EP 239 400 (preferably incorporated herein by
reference, especially regarding the methods for constructing a
variable domain gene) and may be briefly summarized as follows: A
gene encoding a variable domain of a monoclonal antibody of
whatever specificity is cloned. The DNA segments encoding the
framework and hypervariable regions are determined and the DNA
segments encoding the hypervariable regions are removed so that the
DNA segments encoding the framework regions are fused together with
suitable restriction sites at the junctions. The restriction sites
may be generated at the appropriate positions by mutagenesis of the
DNA molecule by standard procedures. Double stranded synthetic
variable region cassettes are prepared by DNA synthesis according
to the sequences given above. These cassettes are provided with
sticky ends so that they can be ligated at the junctions to the
framework by standard protocol for achieving a DNA molecule
encoding an immunoglobulin variable domain.
[0093] Furthermore, it is not necessary to have access to the mRNA
from a producing hybridoma cell line in order to obtain a DNA
construct coding for the monoclonal antibodies of the invention.
Thus, PCT application WO 90/07861 (preferably incorporated herein
by reference, especially with regard to the production of
monoclonal antibodies) gives full instructions for the production
of a monoclonal antibody by recombinant DNA techniques given only
written information as to the nucleotide sequence of the gene.
[0094] The method comprises the synthesis of a number of
oligonucleotides, their amplification by the PCR method, and their
splicing to give the desired DNA sequence.
[0095] Expression vectors comprising a suitable promoter or genes
encoding heavy and light chain constant parts are publicly
available. Thus, once a DNA molecule of the invention is prepared
it may be conveniently transferred in an appropriate expression
vector.
[0096] DNA molecules encoding single chain antibodies may also be
prepared by standard methods, for example, as described in WO
88/1649 (preferably incorporated herein by reference, especially
with regard to the DNA molecules encoding single chain
antibodies).
[0097] In a particular embodiment of the invention, the recombinant
means for the production of some of the binding molecules of the
invention includes first and second DNA constructs as described
below:
[0098] The first DNA construct encodes a heavy chain or fragment
thereof and comprises [0099] a) a first part which encodes the
variable domain of the heavy chain of either antibody 4784,
DNA-4784 V.sub.H (SEQ ID NO: 8), or antibody 4785, DNA-4785 V.sub.H
(SEQ ID NO: 9); this first part starting with a codon encoding the
first amino acid of the variable domain and ending with a codon
encoding the last amino acid of the variable domain, and [0100] b)
a second part encoding a heavy chain constant part or fragment
thereof which starts with a codon encoding the first amino acid of
the constant part of the heavy chain and ends with a codon encoding
the last amino acid of the constant part or fragment thereof,
followed by a non-sense codon.
[0101] Preferably, the second part encodes the constant part of a
human heavy chain, more preferably the constant part of the human
.gamma.4 chain. This second part may be a DNA fragment of genomic
origin (comprising introns) or a cDNA fragment (without
introns).
[0102] The second DNA construct encodes a light chain or fragment
thereof and comprises [0103] a) a first part which encodes the
variable domain of the light chain of either antibody 4784,
DNA-4784 V.sub.L (SEQ ID NO: 10), or antibody 4785, DNA-4785
V.sub.L (SEQ ID NO: 11); this first part starting with a codon
encoding the first amino acid of the variable domain and ending
with a codon encoding the last amino acid of the variable domain,
and [0104] b) a second part encoding a light chain constant part or
fragment thereof which starts with a codon encoding the first amino
acid of the constant part of the light chain and ends with a codon
encoding the last amino acid of the constant part or fragment
thereof followed by a non-sense codon.
[0105] Preferably, the second part encodes the constant part of a
human light chain, more preferably the constant part of the human
.kappa. chain.
[0106] Each of the DNA constructs are placed under the control of
suitable control sequences, in particular under the control of a
suitable promoter. Any kind of promoter may be used, provided that
it is adapted to the host organism in which the DNA constructs will
be transferred for expression. However, if expression is to take
place in a mammalian cell, it is particularly preferred to use the
promoter of an immunoglobulin gene.
[0107] The desired antibody may be produced in a cell culture or in
a transgenic animal. A suitable transgenic animal may be obtained
according to standard methods which include micro injecting into
eggs the first and second DNA constructs placed under suitable
control sequences transferring the so prepared eggs into
appropriate pseudo-pregnant females and selecting a descendant
expressing the desired antibody.
[0108] When the antibody chains have to be produced in a cell
culture, the DNA constructs must first be inserted into either a
single expression vector or into two separate but compatible
expression vectors, the latter possibility being preferred.
[0109] Accordingly, the invention also provides an expression
vector able to replicate in a prokaryotic or eukaryotic cell line
which comprises at least one of the DNA constructs above
described.
[0110] Each expression vector containing a DNA construct is then
transferred into a suitable host organism. When the DNA constructs
are separately inserted on two expression vectors, they may be
transferred separately, i.e. one type of vector per cell, or
co-transferred, this latter possibility being preferred. A suitable
host organism may be a bacterium, a yeast or a mammalian cell line,
this latter being preferred. More preferably, the mammalian cell
line is of lymphoid origin e.g. a myeloma, hybridoma or a normal
immortalized B-cell, but does not express any endogeneous antibody
heavy or light chain.
[0111] It is also preferred that the host organism contains a large
number of copies of the vectors per cell. If the host organism is a
mammalian cell line, this desirable goal may be reached by
amplifying the number of copies according to standard methods.
Amplification methods usually consist of selecting for increased
resistance to a drug, said resistance being encoded by the
expression vector.
[0112] In another aspect of the invention, there is provided a
process for producing a multi-chain binding molecule of the
invention, which comprises (i) culturing an organism which is
transformed with the first and second DNA constructs of the
invention and (ii) recovering an active binding molecule of the
invention from the culture.
[0113] Alternatively, the heavy and light chains may be separately
recovered and reconstituted into an active binding molecule after
in vitro refolding. Reconstitution methods are well-known in the
art; Examples of methods are in particular provided in EP 120 674
or in EP 125 023. Therefore a process may also comprise [0114] (i)
culturing a first organism which is transformed with a first DNA
construct of the invention and recovering said heavy chain or
fragment thereof from the culture and [0115] (ii) culturing a
second organism which is transformed with a second DNA construct of
the invention and recovering said light chain or fragment thereof
from the culture and [0116] (iii) reconstituting in vitro an active
binding molecule of the invention from the heavy chain or fragment
thereof obtained in (i) and the light chain or fragment thereof
obtained in (ii).
[0117] In a similar manner, there is also provided a process for
producing a single chain or single domain binding molecule of the
invention which comprises [0118] (i) culturing an organism which is
transformed with a DNA construct respectively encoding a single
chain or single domain binding molecule of the invention and [0119]
(ii) recovering said molecule from the culture.
[0120] The binding molecules of the invention significantly inhibit
the binding of LINGO-1 to NgR, significantly attenuate the neurite
outgrowth inhibitory activity of adult rat spinal cord myelin at
sub-nM concentrations and significantly increase oligodendrocyte
differentiation in vitro as exemplified below:
FIGURE LEGENDS
[0121] FIG. 1. Effect of Fabs 4784 and 4785 on AP-LINGO-1 binding
to NgR:SH-SY5Y cells NgR:SH-SY5Y cells in suspension are incubated
with either 1 nM AP or AP-LINGO-1 in the absence or presence of 2
.mu.M of the indicated anti-LINGO-1 Fab or anti-hen lysozyme Fab
3207. Bound AP activity on the cells is measured as absorbance at
405 nm after a 30 min incubation with 1-Step.TM. PNPP. The specific
binding of AP-LINGO-1 is calculated as the difference between the
total amount of AP-LINGO-1 binding and the amount of binding with
AP alone. The mean percentage inhibition of specific binding (n=3,
.+-.STD) is calculated as the percentile difference between the
amount of specific binding of AP-LINGO-1 in the presence of Fab
3207 and the presence of an anti-LINGO-1 Fab.
[0122] FIG. 2. Disinhibition of spinal cord myelin by anti-LINGO-1
IgG4 antibodies 4784 and 4785
[0123] A) P7 CGN cells are incubated for 16 hr on wells coated
without spinal cord myelin (no SC, white bars) or wells coated with
spinal cord myelin in the absence (SC, red bars) or presence of
anti-LINGO-1 IgG4 antibodies, a control anti-lysozyme IgG4 antibody
3207 (green bars) or 1 .mu.m of the ROCK inhibitor Y27632 (yellow
bar). ROCK is the secondary messenger in the signaling pathway of
most, it not all, myelin-associated neurite outgrowth inhibitors,
including those which do not signal through the NgR receptor
complex and as such Y27632 treatment is used as a positive control
for the attenuation of the neurite outgrowth inhibitory activity of
spinal cord myelin (FIG. 1). The experiment is performed in three
96 well plates with an SC and no SC condition per plate to which
the effects of the antibodies on that plate are compared and mean
neurite length per neuron (.mu.m) is calculated for 500 neurons per
well in replicates of 10. The percentage inhibition (white text) is
calculated as the percentile difference in mean neurite
length/neuron between cells plated on wells coated with and without
SC. The percentage disinhibition (black italic text) is calculated
as the difference in mean neurite length between cells plated on SC
in the presence and absence of anti-LINGO-1 antibody as a
percentile of the difference between cells plated on wells coated
with and without SC. *p<0.05, **p<0.01 (one way ANOVA,
Holm-Sidak comparison to mean neurite length/neuron for cells
plated on spinal cord myelin in the absence of antibody).
[0124] B) Fluorescent images of a representative field of view of
cells incubated on wells coated without spinal cord myelin (no SC)
and on wells coated with spinal cord myelin in the absence (SC) or
presence of 1 nM control IgG4 3207 or anti-LINGO-1 IgG4 4784. Cells
grown on spinal cord myelin in the presence of 4784 have visibly
longer neurites and more neurites per cell than those grown in the
absence of antibody or presence of the control antibody 3207.
[0125] FIG. 3. Disinhibition of spinal cord myelin by anti-LINGO-1
IgG4 antibodies II
[0126] A) P7 CGN cells are incubated for 8 hr on wells coated
without spinal cord myelin (no SC, white bars) or wells coated with
spinal cord myelin in the absence (SC, red bars) or presence of
anti-LINGO-1 IgG4 antibodies or a control anti-lysozyme IgG4
antibody 3207. The experiment is performed in three 96 well plates
with an SC and no SC condition per plate to which the effects of
the antibodies on that plate are compared and mean neurite length
per neuron (.mu.m) is calculated for 500 neurons per well in
replicates of 10. The percentage inhibition (white text) and
disinhibition (black italic text) is calculated as above.
**p<0.01 (one way ANOVA, Holm-Sidak comparison to mean neurite
length/neuron for cells plated on spinal cord myelin in the absence
of antibody).
[0127] FIG. 4. Disinhibition of spinal cord myelin by anti-LINGO-1
IgG4 antibodies III
[0128] P7 CGN cells are incubated for 8 hr on wells coated without
spinal cord myelin (no SC) or wells coated with spinal cord myelin
in the absence (SC) or presence of the indicated concentrations of
anti-LINGO-1 IgG4 antibodies 4784 or 4785, a control anti-lysozyme
IgG4 antibody 3207 or 1 .mu.M Y27632 (ROCK). The experiment is
performed in three 96 well plates with an SC and no SC condition
per plate to which the effects of the antibodies on that plate are
compared and mean neurite length per neuron (.mu.m) is calculated
for 500 neurons per well in replicates of 10. The percentage
inhibition (white text) and disinhibition (black italic text) is
calculated as above. *p<0.05, **p<0.01 (one way ANOVA,
Holm-Sidak comparison to mean neurite length/neuron for cells
plated on spinal cord myelin in the absence of antibody).
[0129] FIG. 5. Anti-LINGO-1 antibodies significantly increase the
differentiation of immature oligodendrocytes
[0130] A) Freshly isolated OPCs are treated with 100 nM 4784, 4785
or control IgG4 3207 for 3 days in DMEM/CNTF/T3 medium followed by
staining with the anti-O4 antibody to visualise immature and mature
oligodendrocytes (larger, more diffuse labeling) and the nucleic
acid dye DAPI (4',6-diamidin-2'-phenyl-indol-dihydrochloride) to
visualise cell nuclei (smaller circular dots). Oligodendrocytes
bearing highly arborised and extended processes and myelin
sheet-like structures are considered to have a mature morphology
and are indicated with white arrows. Anti-LINGO-1 antibody
treatment results in an increase in the proportion of O4-positive
cells with a mature morphology whereas treatment with control IgG4
3207 has no effect.
[0131] B) The proportion of total (left graph) and mature (right
graph) oligodendrocytes is quantified in three independent
experiments (1,2,3). The left bar graph depicts the percentage of
DAPI-stained nuclei associated with O4-staining and the right bar
graph depicts the percentage of O4-positive cells with a mature
morphology (mean of triplicates+STD). In each bar graph, the
leftmost bar is with no treatment, the second to left bar Control
with Control IgG, the next represents treatment with 4784 and the
rightmost treatment with 4785. Anti-LINGO-1 antibodies have no
effect on the proportion of cells that are oligodendrocytes but
significantly increase the proportion of oligodendrocytes with a
mature morphology. * p<0.05, ** p<0.01, one-way ANOVA with a
Holm-Sidak comparison to the proportion of mature oligodendrocytes
in the presence of the control IgG4 3207.
[0132] FIG. 6. Anti-LINGO-1 antibodies downregulate cell surface
LINGO-1
[0133] A) Untransfected CHO-K1 or CHO-K1-hLINGO-1 cells are
incubated at 37.degree. C. for 24 hrs with 100 nM 4784, 4785 and
3207 and LINGO-1 detected at the cell surface by a further
incubation at room temperature for 30 min with the anti-V5
antibody. The cells are fixed with 4% PFA, blocked with BSA and
bound anti-V5 antibody detected using an anti-mouse-IgG (Fc
specific)-POD conjugate that is subsequently developed using a
1-Step.TM. Turbo TMB ELISA kit. The absorbance at 450 nm is taken
as a measure of the amount of LINGO-1 at the cell surface (mean of
triplicates.+-.STD). A very low level of anti-V5 antibody binding
is observed to untransfected CHO-K1 cells. Incubation of
CHO-K1-hLINGO-1 cells with anti-LINGO-1 antibodies but not the
control IgG4 3207 result in a significant reduction in the amount
of LINGO-1 at the cell surface ** p<0.01, one-way ANOVA with a
Holm-Sidak comparison to the absorbance following incubation with
the control IgG4 3207.
[0134] B) Cell surface proteins on untransfected CHO-K1 or
CHO-K1-hLINGO-1 cells are biotinylated at 4.degree. C. and the
cells are incubated at 37.degree. C. for the indicated times with
or without 100 nM 4784, 4785 and 3207. At the end of the incubation
period, LINGO-1 is precipitated from the cell lysate using the
anti-V5 antibody coupled to agarose beads and biotinylated (cell
surface) LINGO-1 detected by Western blot analysis using an
anti-biotin antibody. No signal is detected for biotinylated
LINGO-1 in untransfected CHO-K1 cells. Incubation of
CHO-K1-hLINGO-1 cells with anti-LINGO-1 antibodies increases the
rate of degradation of cell surface LINGO-1.
[0135] FIG. 7. Characterization of anti-LINGO-1 Fabs by ELISA
[0136] Values for ELISA analyses are given as mean values of
relative fluorescence units (RFU). The binding affinities of these
clones are characterized by FACS saturation assays.
[0137] The present invention also provides the use of the binding
molecules of the invention in the promotion of axonal
regeneration/plasticity of a mammalian nervous system, in
particular the human nervous system.
[0138] The invention also provides a method of promoting axonal
regeneration/plasticity of a mammalian nervous system, in
particular human nervous system which comprises administering an
effective amount of the binding molecules of the invention to a
patient in need of such treatment.
[0139] The invention also provides a pharmaceutical composition for
promoting axonal regeneration/plasticity of a mammalian nervous
system, in particular human nervous system which comprises the
binding molecules of the invention and a pharmaceutically
acceptable carrier or diluent.
[0140] In particular, the binding molecules of the invention are
useful for promoting axonal regeneration and plasticity after CNS
injury (the term injury, in the present application, refers
especially to injury caused by mechanical or chemical effects or
due to diseases or disorders that e.g. lead to degeneration of
neurons, especially their structure or form, e.g. in neurological
diseases such as Alzheimer's or Parkinson's Disease or other
disorders or diseases mentioned below). Thus the molecules of the
invention have a wide utility in particular for human subjects. For
example the binding molecule of the invention are useful in the
treatment of various diseases of the peripheral (PNS) and central
(CNS) nervous system, i.e. more particularly in neurodegenerative
diseases such as Alzheimer's disease, Parkinson's disease,
Amyotrophic lateral sclerosis (ALS), Lewy like pathologies or other
dementia in general, diseases following cranial, cerebral or spinal
trauma and stroke. Furthermore, given that LINGO-1 is a negative
regulator of myelination, the binding molecules of the invention
are useful for promoting remyelination in concert with promoting
axonal regeneration/plasticity in demyelinating diseases that
include, but are not limited to, multiple sclerosis, monophasic
demyelination, encephalomyelitis, multifocal leukoencephalopathy,
panencephalitis, Marchiafava-Bignami disease, pontine myelmolysis,
adrenoleukodystrophy, Pelizaeus-Merzbacher disease, Spongy
degeneration, Alexander's disease, Canavan's disease, metachromatic
leukodystrophy and Krabbe's disease. In one example, cells which
express the binding molecules of the invention may be transplanted
to a site spinal cord injury to facilitate axonal growth throughout
the injured site. Such transplanted cells would provide a means for
restoring spinal cord function following injury or trauma. Such
cells could include olfactory ensheathing cells and stem cells of
different lineages of fetal nerve or tissue grafts.
[0141] In addition, the binding molecules of the invention are
useful for the treatment of degenerative ocular disorders which may
directly or indirectly involve the degeneration of retinal or
corneal cells including ischemic retinopathies in general, anterior
ischemic optic neuropathy, all forms of optic neuritis, age-related
macular degeneration, diabetic retinopathy, cystoid macular edema
(CME), retinitis pigmentosa, Stargardt's disease, Best's
vitelliform retinal degeneration, Leber's congenital amaurosis and
other hereditary retinal degenerations, pathologic myopia,
retinopathy of prematurity, and Leber's hereditary optic
neuropathy, the after effects of corneal transplantation or of
refractive corneal surgery, and herpes keratitis.
[0142] Furthermore, the binding molecules of the invention are
useful for the treatment of psychiatric conditions, particularly
schizophrenia and depression.
[0143] For these indications, the appropriate dosage will, of
course, vary depending upon, for example, the particular molecule
of the invention to be employed, the mode of administration and the
nature and severity of the condition being treated. In general, the
dosage preferably will be in the range of 1 .mu.g/kg/day to 1
mg/kg/day. The binding molecules of the invention are conveniently
administered by pumps or injected as therapeutics at the lesioned
site or near it, e.g. they can be administered directly into the
CNS intracranially or into the spine intrathecally to the lesioned
site. However, systemic administration is not excluded here. The
binding molecules of the invention can be provided alone, or in
combination, or in sequential combination with other agents. For
example, the binding molecules of the invention can be administered
in combination with anti-Nogo-A antibodies or anti-inflammatory
agents such as but not limited to corticosteroids following stroke
or spinal cord injury as a means for blocking further neuronal
damage and inhibition of axonal regeneration, neurotrophic factors
such as NGF, BDNF or other drugs for neurodegenerative diseases
such as Exelon.TM. or Levodopa. Other suitable combination partners
for the treatment of stroke are Alteplase and Desmoteplase (DSPA,
e.g. disclosed in WO90/09438). In one embodiment, the present
invention provides a combination comprising a binding molecule of
the invention and Desmoteplase, in particular for the treatment of
stroke as well as pharmaceutical compositions comprising said
combination. As used herein, two agents are said to be administered
in combination when the two agents are administered simultaneously
or are administered independently in a fashion such that the agents
will act at the same time.
[0144] The structure of the active ingredients identified by code
nos., generic or trade names may be taken from the actual edition
of the standard compendium "The Merck Index" or from databases,
e.g. Patents International (e.g. IMS World Publications) or other
databases provided by IMS Health. The corresponding content thereof
is hereby incorporated by reference. Any person skilled in the art
is fully enabled to identify the active ingredients and, based on
these references, likewise enabled to manufacture and test the
pharmaceutical indications and properties in standard test models,
both in vitro and in vivo.
[0145] Pharmaceutical compositions of the invention may be
manufactured in conventional manner. E.g. a composition according
to the invention comprising the molecules of the invention is
preferably provided in lyophilized form. For immediate
administration it is dissolved in a suitable aqueous carrier, for
example sterile water for injection or sterile buffered
physiological saline.
[0146] To aid in making up suitable compositions, the binding
molecules of the invention and optionally a second drug enhancing
the effect of the binding molecules of the invention, may be
packaged separately within the same container, with instructions
for mixing or concomitant administration. Optional second drug
candidates are provided above.
[0147] The synergistic effect of a combination of the binding
molecules of the invention and growth factors such as NGF may be
demonstrated in vivo by the spinal cord injury models.
[0148] The invention will be more fully understood by reference to
the following examples. They should not, however, be construed as
limiting the scope of the invention.
[0149] The monoclonal antibodies of attention in the Examples are
binding molecules according to the present invention containing for
antibody 4784 the variable part of the light chain (SEQ ID NO: 4)
and the variable part of the heavy chain (SEQ ID NO: 5) and
comprising for 4785 the variable part of the light chain (SEQ ID
NO: 6) and the variable part of the heavy chain (SEQ ID NO: 7).
[0150] The following abbreviations are used:
AP human placental alkaline phosphatase CDR complementarity
determining region cDNA complementary DNA ELISA enzyme linked
immuno-sorbant assay FACS fluorescence activated cell sorting FBS
foetal bovine serum HCMV human cytomegalovirus promoter IgG
immunoglobulin isotype G PBS phosphate-buffered saline PCR
polymerase chain reaction PFA paraformaldehyde PNPP
para-nitrophenyl phosphate
Example 1
Generation of CHO-K1 Cells Expressing Full Length Rat, Cynomolgus
Monkey or Human LINGO-1 and Human LINGO-2
[0151] A human cDNA library is generated by RT-PCR of universal
human reference RNA (Stratagene) using random and oligo dT primers.
A cynomolgus monkey brain cDNA library is generated by RT-PCR of
polyA RNA isolated from frozen cynomolgus monkey brain using random
and oligo dT primers. A Marathon-ready rat brain cDNA library is
obtained from Clontech. cDNA encoding the mature sequence (residues
34-614) of human LINGO-1 (SEQ ID NO: 27), cynomolgus monkey LINGO-1
(SEQ ID NO: 28) and rat LINGO-1 (SEQ ID NO: 29) flanked by 5''-XbaI
and 3'-XhoI sites is PCR amplified from the respective library
using the forward primer DM14,
5''-CTACGTCTAGAACGGGCTGCCCGCCCCGCT-3'' (SEQ ID NO: 30), and reverse
primer DM15, 5''-GGTTTCTCGAGTCATATCATCTT CATGTTGAACTTGCGG-3'' (SEQ
ID NO: 31). The PCR product is cleaved with XbaI and XhoI and
inserted into the respective sites of the vector pSecTag2-V5 (SEQ
ID NO: 32) to generate hLINGO-1-pSecTag2-V5, cmLINGO-1-pSecTag2-V5
and rLINGO-1-pSecTag2-V5, respectively. The predicted protein
product is the mature sequence of LINGO-1 fused at the N-terminus
to a 14 amino acid residue V5 epitope tag via a 2 amino acid
residue linker. cDNA encoding the mature sequence (residues 26-606)
of human LINGO-2 (SEQ ID NO: 33) flanked by 5''-XbaI and 3''-XhoI
sites is PCR amplified from a Marathon-ready human brain cDNA
library (Clontech) using the forward primer DM16,
5'-CTACGTCTAGAATTGGCTGCCCCGCTCGCT-3'' (SEQ ID NO: 34), and reverse
primer DM17, 5''-GGTTTCTCGAGTCAAATCATTTTCATGTTGAAC CTCCTG-3' (SEQ
ID NO: 35). The PCR product is cleaved with XbaI and XhoI and
inserted into the respective sites of the vector pSecTag2-V5 to
generate hLINGO-2-pSecTag2-V5. The predicted protein product is the
mature sequence of LINGO-2 fused at the N-terminus to a 14 amino
acid residue V5 epitope tag via a 2 amino acid residue linker.
CHO-K1 cells stably expressing human LINGO-1 (CHO-K1-hLINGO-1),
cynomolgous LINGO-1 (CHO-K1-cmLINGO-1), rat LINGO-1
(CHO-K1-rLINGO-1) and human LINGO-2 (CHO-K1-hLINGO-2) are generated
by transfection of cells with hLINGO-1-pSecTag2-V5,
cmLINGO-1-pSecTag2-V5, rLINGO-1-pSecTag2-V5 and
hLINGO-2-pSecTag2-V5, respectively, using lipofectamine-2000
(Invitrogen) according to the manufacturer's instructions. Stably
expressing transfectants are selected with 1 mg/ml zeocin
(Invivogen) and single clones isolated either by serial dilution
into 96-well plates or by using clonal rings. Expression of the
constructs on the cell surface is confirmed by immunofluorescent
analysis using an anti-V5 antibody (InvitroGen).
Example 2
Generation and Expression of Human LINGO-1-Fc and Human
LINGO-1.DELTA.LRR-Fc
[0152] A MGC mRNA coding for human LINGO-1 (clone MGC:17422
IMAGE:4214343) is used as template for PCR amplification. The
extracellular domain (ECD) preceded by the natural signal sequence
(aa1-550) of human LINGO-1 is amplified by PCR with the Pwo1
polymerase (Roche Diagnostics) and with primers which added a
HindIII restriction site and a Kozak consensus sequence at the 5'
end of the target sequence and an XhoI restriction site immediately
after the last codon of the target sequence at the 3' end. The PCR
product is digested with HindIII and XhoI, gel purified and
inserted into plasmid pRS5a-IgG (SEQ ID NO: 36) previously digested
with the same enzymes. The accuracy of the inserted sequence,
complete Fc and flanking regions in the resulting expression clone
(natleader-hsLINGO-1-Fc/pRS5a, SEQ ID NO: 37) is confirmed by DNA
sequencing.
[0153] The same MGC clone serves as template for the construction
by gene SOEing of the expression plasmid for human LINGO-1 lacking
the LRR domain (aa34-65+aa354-550). The N-terminal region of human
ECD LINGO-1 (aa34-65) is amplified by PCR with primers extending
the 5' end with a partial sequence coding for an heterologous
secretion signal fused to mature LINGO-1 and adding, at the 3' end,
a sequence coding for the first seven amino acids of the C-terminal
fragment. The C-terminal region of human ECD LINGO-1 (aa354-550) is
amplified by PCR with primers extending the 5' end with a sequence
coding for the last seven amino acids of the N-terminal fragment
and adding, at the 3' end, an XhoI site immediately after the last
codon of the target sequence. The two PCR products are gel
purified, mixed and serves as template for a second PCR
amplification using at the 5' end a primer which adds a HindIII
restriction site, a Kozak consensus sequence and completes the
herologous secretion signal sequence and, at the 3' end, the
external primer previously used to amplify the C-terminal fragment.
The PCR product is digested with HindIII and XhoI, gel purified and
inserted into plasmid pRS5a-IgG previously digested with the same
enzymes. The accuracy of the inserted sequence, complete Fc and
flanking regions in the resulting expression clone
(Igleader-hsLINGO-1-.DELTA.LRR-Fc/pRS5a, SEQ ID NO: 38) is
confirmed by DNA sequencing.
[0154] As an initial expression evaluation both constructs are
tested in small scale experiments. HEK.EBNA cells (Invitrogen,
previous cat.no. R620-07) are cultivated in attached mode on tissue
culture flasks in Dulbecco's Modified Eagle Medium (DMEM) buffered
with 25 mM Hepes (Gibco/Life Technologies cat.no. 42430-025) and
additionally enriched with 10% fetal calf serum; the cultures are
maintained at 37.degree. C. and 5% CO.sub.2 in humidified
atmosphere. For small scale transfection experiments,
4.times.10.sup.5 cells are seeded one day prior to transfection
into poly-D-lysine-coated 6-wells (plates). Transfections are
performed using 3 .mu.g of plasmid DNA and 6 .mu.l of
Lipofectamine.sup.2000 reagent (Invitrogen cat.no. 11668-019) per
well, essentially as described by the vendor. Three days
post-transfection the cell supernatants are harvested and the
cell-free supernatant is subjected to protein analysis, i.e. to
immuno-affinity HPLC analysis on Protein G columns. Titers ranging
between 8 mg/l for construct natleader-hsLINGO-1-Fc/pRS5a and 40
mg/l for construct Igleader-hsLINGO-1-.DELTA.LRR-Fc/pRS5a are
determined. Subsequently, for both plasmids large-scale plasmid
preps are prepared to enable transient transfections on the
multi-litre scale in HEK.EBNA suspension cultures.
[0155] For production of natleader-hsLINGO-1-Fc on enlarged scale,
2.9 L of HEK.EBNA cell culture at a density of 1.4.times.10.sup.6
cells/ml is mixed with 1.1 L DNA:PEI solution (1 .mu.g DNA:2 .mu.g
PEI per ml). Following incubation of cells for 4 hrs, the culture
is fed with 4 L of ExCell VPRO medium (SAFC, previously JRH,
Lenexa, Kans.). The cell culture supernatant is harvested after 6
days of cultivation and concentrated by diafiltration down to 1-L
using a disposable Hemoflow F10HPS filter with a 10 kDa cut-off
(Fresenius Medical Care, Germany).
[0156] The second relevant protein production run to generate
Igleader-hsLINGO-1-.DELTA.LRR-Fc protein is done in a similar
fashion. Details on large-scale transfection, DNA:PEI ratio, cell
densities, feeding and harvest are exactly the same as described
above.
a) natleader-hsLINGO-1-Fc
[0157] 1 L concentrate (from 8 L culture supernatant) is
chromatographed on 20 ml Protein A Sepharose. After base-line
washing with 100 mM NaPi, pH 7.3, bound material is eluted with 50
mM citrate, 140 mM NaCl, pH 2.7, neutralized and sterile filtered.
The eluted fraction is further concentrated and gel filtered on
Superdex 75 in PBS yielding 8.2 mg product at a concentration of
1.2 mg/ml.
b) Igleader-hsLINGO-1-.DELTA.LRR-Fc
[0158] 1 L concentrate (from 8 L culture supernatant) is
chromatographed on 20 ml Protein A Sepharose. After base-line
washing with 100 mM NaPi, pH 7.3, bound material is eluted with 50
mM citrate, 140 mM NaCl, pH 2.7, neutralized and sterile filtered
yielding 52.5 mg product at a concentration of 1.5 mg/ml.
[0159] The purified proteins are extensively characterized by
N-terminal sequencing and by MALDI peptide mass analysis after
reduction/alkylation and trypsin digestion.
Example 3
AP-LINGO-1 Binding Assay
[0160] Blocking the binding of LINGO-1 to NgR is expected to
prevent the signaling of three myelin-associated inhibitors of
neurite outgrowth, namely Nogo-66, MAG and OMgp, and hence
attenuate the neurite outgrowth inhibitory activity of CNS myelin
thus leading to increased axonal regeneration/plasticity and
improved functional recovery following acute CNS injury. To
demonstrate that an anti-LINGO-1 antibody blocks the binding of
LINGO-1 to NgR, an assay can be used which measures the binding of
human placental alkaline phosphatase (AP)-tagged rat LINGO-1
ectodomain (AP-LINGO-1) to SH-SY5Y cells stably expressing NgR
(NgR-SH-SY5Y, Walmsley et. al. (2004) J Cell Sci 117, 4591-4602).
cDNA encoding the majority of the rat LINGO-1 ectodomain (residues
34-532) flanked by 5'-Xho I and 3'-Xba I sites is PCR amplified
from rLINGO-1-pSecTag2-V5 using the forward primer DM22,
5''-GGTTATCTCGAGACCGGCTGCCCGCCCC-3' (SEQ ID NO: 24), and reverse
primer DM23, 5''-GGCCCTTCTAGATCACTCGCCTGGCTGGTTGGAGATG-3'' (SEQ ID
NO: 25). The PCR product is cleaved with XhoI and XbaI and inserted
into the respective sites of the vector APtag-5-NHIS (SEQ ID NO:
26) to generate APtag-5-NHIS-solrLINGO-1. The predicted protein
product is the majority of the rat LINGO-1 ectodomain fused at the
N-terminus to residues 23-511 of human placental alkaline
phosphatase via a 3 amino acid residue linker. HEK293T cells are
transfected with APtag-5-NHIS-solrLINGO-1 using
lipofectamine.sup.2000 according to the manufacturer's
instructions. The transfection medium is removed 4 hrs after
transfection and replaced with OptiMEM I without phenol red
(Invitrogen). Medium is harvested after 24 hrs, replaced and
harvested again after another 24 hrs. The medium is clarified by
centrifugation at 13000.times.g for 5 min and the supernatant
concentrated around 15-fold using a Centriprep filter device
(Millipore) according to the manufacturer's instructions. AP
activity of the concentrated supernatant is measured using
1-Step.TM. PNPP (Pierce) as change in absorbance at 405 nm over
time and transformed to a concentration using the following
equation (applies for a 96 well plate format with 200 .mu.l
PNPP/well):
Concentration of AP - fusion ( n M ) = Change in absorbance ( mAU /
min ) 7.945 .times. volume of sample added to PNPP ( .mu. l )
##EQU00001##
[0161] Concentrated supernatant is subjected to SDS-PAGE gel
electrophoresis and Western blotted as described (Walmsley et. al.
(2004) J Cell Sci 117, 4591-4602). AP-LINGO-1 is detected with 0.1%
(v/v) anti-penta-histidine antibody (Qiagen) followed by 0.02%
(v/v) peroxidase-conjugated anti-mouse IgG antibody (Sigma) using
the ECL.TM. system (GE Healthcare). AP-LINGO-1 is visualised as a
band of approximately 110 kDa, similar to its predicted molecular
weight of 112 kDa. No N-terminal degradation products are observed.
NgR:SH-SY5Y cells at 50% confluency are harvested with enzyme-free
dissociation buffer (Invitrogen) to preserve cell surface proteins
such as NgR. 1 nM AP, 1 nM AP-LINGO-1 or 1 nM AP-LINGO-1 in the
presence of 2 .mu.M anti-LINGO-1 Fab or a control Fab 3207 against
lysozyme from hen egg white is pre-incubated for 30 min in OptiMEM
(Invitrogen) and subsequently incubated with constant agitation for
1.5 hr with NgR:SH-SY5Y cells in suspension. Cells are washed 6
times in HBH (20 mM HEPES pH 7.4/1% bovine serum albumin in Hanks
balanced saline) and fixed in 4% paraformaldehyde (PFA)/5% sucrose
in PBS for 15 min. Following inactivation of endogenous AP activity
by incubation at 65.degree. C. for 1 hr in 20 mM HEPES pH 7.4 in
Hanks balanced saline, cell-bound AP activity is quantified as
absorbance at 405 nm after a 30 min incubation with 1-Step.TM. PNPP
(Pierce) according to the manufacturer's instructions.
[0162] The Fabs are used at a concentration of 2 .mu.M in order to
saturate AP-LINGO-1 with bound Fab and thus minimise the influence
of their affinities on their ability to inhibit binding. The reason
for this is to exclude the possibility of prematurely discarding
Fabs from further studies which fail to inhibit binding due to
their low affinity rather than the position of their binding site
as the affinity of such Fabs could be increased at later stages by
affinity maturation and IgG4 conversion. 1 nM AP-LINGO-1 is
pre-incubated with either the control Fab 3207 or anti-LINGO-1 Fabs
4784 and 4785 and then allowed to bind in the presence of the Fab
to NgR:SH-SY5Y cells in suspension (FIG. 1). The percentage
inhibition in specific AP-LINGO-1 binding in the presence of the
anti-LINGO-1 Fabs is normalized to that for Fab 3207. 4784 and 4785
give a significant inhibition (p<0.01, one way ANOVA, Holm-Sidak
comparison to specific binding of AP-LINGO-1 in presence of control
Fab 3207) of AP-LINGO-1 binding to the cells.
[0163] Blocking the binding of LINGO-1 to NgR is predicted to
prevent the signaling of the myelin-associated inhibitors Nogo-66,
MAG and OMgp leading to a reduction in the neurite outgrowth
inhibitory activity of CNS myelin. In that regard, 4784 and 4785
Fabs are converted to the final IgG4 format (see Example 8) and
assessed for their ability to attenuate the inhibition of neurite
outgrowth from postnatal day 7 rat cerebellar granule neurons grown
on adult rat spinal cord myelin.
Example 4
Neurite Outgrowth Inhibition Assay
[0164] The most relevant in vitro assay to predict the effect of
anti-LINGO-1 antibodies on axonal regeneration/plasticity in vivo
is their ability to attenuate the neurite Outgrowth inhibitory
activity of CNS myelin. In this assay, postnatal day 7 rat
cerebellar granule neurons (CGN) are grown in wells coated with
whole spinal cord myelin extracted from adult rats and neurite
outgrowth quantified by an automated ArrayScan.RTM. HCS Reader
(Cellomics).
[0165] The disinhibitory activity of anti-LINGO-1 IgG4 antibodies
4784 and 4785 is assessed in the said neurite outgrowth assay (FIG.
2).
[0166] Fresh rat spinal cord tissue from adult rats is homogenized
in 3 volumes (w/v) extraction buffer (60 mM Chaps, 20 mM Tris pH
8.0, 1 mM EDTA, protease inhibitor cocktail), incubated for 30 min
at 4.degree. C. and clarified by centrifugation at 170000.times.g
for 30 min at 4.degree. C. Each well in a 96 well plate is coated
with 5 .mu.l nitrocellulose in MeOH (5 cm.sup.2 nitrocellulose in
12 ml MeOH), air dried and coated with 100 .mu.l 5 .mu.g/ml
poly-D-lysine by incubation for 4 hr at 37.degree. C. Following
three washes in water, the plates are air dried for 1 hr and then
coated with 60 .mu.g/cm.sup.2 spinal cord extract by incubation
overnight at 37.degree. C. CGN cells are freshly purified from
trypsin dissociates of postnatal day 7 rat cerebellar tissue as
described previously (Schweigreiter et al., 2004). Western blot
analysis to detect LINGO-1 is performed on lysates from CHO-K1
cells expressing V5-tagged rat LINGO-1 or P7 CGN cells using 2
.mu.g/ml (or 13.3 nM) anti-LINGO-1 polyclonal antibody (Upstate)
followed by 0.02% (v/v) peroxidase-conjugated anti-rabbit IgG
antibody (Sigma). CGN cells (35000 cells/well) are incubated for 30
min at 37.degree. C. on wells coated without or with spinal cord
myelin prior to the addition of either 0-100 nM anti-LINGO-1 IgG4
antibody or the control 3207 IgG4 antibody. Following an 8-16 hr
incubation at 37.degree. C., cells are fixed with 4% PFA and
stained with Hoechst 3342 (Invitrogen) for visualisation of the
nucleus and anti-.beta.-tubulin III antibody (R&D Systems)
followed by an Alexa Fluor 546-conjugated anti-mouse IgG antibody
(Invitrogen) to specifically visualize neurons. Parameters of
neurite outgrowth are determined using an ArrayScan.RTM. HCS Reader
(Cellomics). ArrayScan.RTM. II automatically locates, focuses and
exposes fields of cells within a 96-well microtiter plate.
ArrayScan.RTM. consists of a high-resolution optical system, a
multiple bandpass emission filter with matched single band
excitation filter (XF100), a CCD camera with frame grabber, and
proprietary applications software. In this assay, the Extended
Neurite Outgrowth Bioapplication is used.
[0167] An excitation filter wheel and multiple bandpass emission
filters are used to enable multichannel imaging of fluorescence
from two fluorophores in the same cells. Bandpass images of Hoechst
33342-labelled nuclei are acquired to identify discrete cells, and
bandpass images of Alexa Fluor 488 are then acquired to identify
the extent of cells labeled with anti-tubulin antibody (using a
secondary conjugated to Alexa Fluor 488). Inappropriate bodies
within cells are automatically excluded from the analysis, so that
only overlapping Hoechst and beta-tubulin cell bodies are analyzed.
Dual emission images are acquired for 5 discrete 350 .mu.m.sup.2
fields in each well of the plate. Using a 10-x objective, this
results in 400-500 cells per well analyzed. The Extended Neurite
Outgrowth Bioapplication then reports several quantitative measures
of neuronal morphology for single cells, including neurite length
number of neurites per cell, cell body area, and branch and cross
points. The mean neurite length per neuron (.mu.m) is calculated
for 500 neurons per well in replicates of 10.
[0168] In the above neurite outgrowth assay, the anti-LINGO-1 IgG4
antibodies 4784 and 4785 are disinhibitory at 1 and 10 nM, whereas
the control IgG4 against lysozyme gives no disinhibition at both
concentrations (FIG. 2). The mean length of neurites per neuron on
spinal cord myelin in the presence of 4784 and 4785 at both
concentrations is statistically higher than that in the absence of
antibody. The greater level of disinhibition achieved with the ROCK
inhibitor Y27632 compared to the anti-LINGO-1 antibodies 4784 and
4785 is expected as this compound inhibits the signaling pathways
of additional myelin-associated neurite outgrowth inhibitors other
than those that signal through the NgR receptor complex.
[0169] To confirm the above results, the neurite outgrowth assay is
repeated (FIG. 3). Again, the anti-LINGO-1 antibodies 4784 and 4785
are disinhibitory at 1 nM and 10 nM, whereas the control IgG4
against lysozyme gives no disinhibition at both concentrations. The
mean length of neurites per neuron on spinal cord myelin in the
presence of 4784 and 4785 at both concentrations is statistically
higher than that in the absence of antibody.
[0170] To further establish the potency of the anti-LINGO-1
antibodies 4784 and 4785, the effect on neurite outgrowth
inhibition of sub-nM concentrations of the antibody is assessed
(FIG. 4). 4784 and 4785 give a significant disinhibition (38-51%
and 51-57%, respectively) of spinal cord myelin at concentrations
as low as 0.1 nM, whereas the control anti-lysozyme antibody has no
effect. Again, the ROCK inhibitor Y27632 gives a higher degree of
disinhibition (65-74%) than the anti-LINGO-1 IgG4 antibodies as
expected.
Example 5
Primary Oliqodendrocyte Differentiation Assay
[0171] Blockade of LINGO-1 function by genetic means or by
treatment with a receptor-body antagonist has been reported to
increase the proportion of mature oligodendrocytes arising from
purified OPC cultures (Mi et al. (2005) Nat Neurosci 8, 745-751).
To assess the ability of anti-LINGO-1 antibodies to block LINGO-1
function in OPC cultures and promote oligodendrocyte maturation,
freshly isolated rat OPCs are incubated with 4784, 4785 or control
IgG4 3207 for 3 days in DMEM/CNTF/T3 medium followed by staining
with the anti-04 antibody to label both immature and mature
oligodendrocytes (FIG. 5). The degree of oligodendrocyte maturation
is measured as the proportion of O4-positive cells exhibiting a
mature morphology.
[0172] Enriched populations of OPCs are isolated from OFA P3 rats:
Briefly, the brain is dissected and the telencephalons are placed
in ice-cold Hank's buffered saline solution (HBSS, Invitrogen)
containing 0.15% MgSO.sub.4. The tissue is incubated with 1:1
HBBS/trypsin-EDTA (Invitrogen) and 100 .mu.g/ml DNAse I (Roche) for
10 min at 37.degree. C. and the trypsin inactivated by addition of
FCS (Invitrogen) to a final concentration of 10%. The tissue
suspension is centrifuged at 890 rpm for 10 min and the pellet
resuspended in Basal Medium Eagle (BME, Invitrogen) with 10% horse
serum (Invitrogen). The suspension is filtered through a 40 .mu.m
cell strainer (BD Falcon) and the cells plated on poly-D-lysine
pre-coated 80 cm.sup.2 tissue culture flasks (BD Falcon) at 1 brain
per flask. Cells are cultivated at 37.degree. C. for 11 days in
BME/10% horse serum. Microglial cells are killed by adding 5 mM
L-leucine-methyl esther and the flasks are agitated by shaking at
140 rpm for 2 hrs. OPCs are harvested by shaking the flasks
overnight at 200 rpm at 37.degree. C. and any astrocytes remaining
in the supernatant are further separated from the OPCs by
pre-attachment for 2 hrs at 37.degree. C. on 10 cm bacterial
culture dishes. Non-adherent cells are collected, centrifuged for
10 minutes at 890 rpm and plated at approximately 3.times.10.sup.4
cells/well in poly-D-lysine-coated 8-well chamber slides (BD
Falcon). Cultures are maintained for 3 days in either in
DMEM/T3/CNTF medium consisting of DMEM (Invitrogen) containing 10
ng/ml Ciliary Neurotrophic Factor (R&D Systems) and 15 nM
Triiodothyronine (Sigma) or in SATO medium consisting of DMEM
(Invitrogen) containing 10 .mu.g/ml transferrin (Sigma), 10
.mu.g/ml insulin (Sigma), 100 .mu.M putrescine (Sigma), 200 nM
progesterone (Sigma), 520 nM thyroxine (Sigma), 500 .mu.M
Triiodothyronine (Sigma), 220 nM sodium selenite (Sigma), 25
.mu.g/ml gentamycin (Sigma) and 1% HS (Invitrogen). To assess the
purity of the cultures with respect to the oligodendrocyte lineage,
the percentage of cells that are stained with the anti-O4 antibody
is quantified after 7 days of culture in SATO medium. Typically,
80-95% of the cells are stained with the anti-O4 antibody
demonstrating that the majority of the cells in the culture are of
the oligodendrocyte lineage.
[0173] To assess oligodendrocyte maturation based on
oligodendrocyte morphology, freshly isolated OPC cultures are
incubated in DMEM/T3/CNTF medium for 3 days in the absence or
presence of 100 nM 4784, 4785 or control IgG4 3207 followed by
staining with the anti-O4 antibody to label both immature and
mature oligodendrocytes and DAPI to label cell nuclei. O4-positive
cells with clearly defined short processes are considered to
represent immature oligodendrocytes whereas O4-positive cells
bearing extended and highly arborised processes with myelin
sheet-like structures are considered to represent mature
oligodendrocytes. The proportion of O4-positive cells with a mature
morphology is quantified for around 300-1300 cells in triplicate
per treatment and significance determined using one-way ANOVA with
a Holm-Sidak comparison to the proportion of mature
oligodendrocytes in the presence of the control IgG4 3207. To
assess the effect of the antibody treatment on the proportion of
total (immature and mature) oligodendrocytes in the culture, the
proportion of DAPI nuclei associated with O4-staining is
quantified.
[0174] In three independent experiments, treatment with the
anti-LINGO-1 antibodies 4784 and 4785 significantly increases the
proportion of oligodendrocytes with a mature morphology as
represented by cells bearing highly arborised processes that extend
over a wide area and myelin sheet-like structures (FIG. 5).
Treatment with the control IgG4 antibody 3207 has no effect on the
proportion of mature oligodendrocytes in the culture. The
proportion of DAPI-stained nuclei associated with O4-staining is
similar for all treatments, demonstrating that anti-LINGO-1
antibodies have no effect on the proportion of cells corresponding
to both immature and mature oligodendrocytes.
[0175] As anti-LINGO-1 antibody treatment has no effect on the
proportion of total oligodendrocytes, the increase in the
proportion of mature oligodendrocytes most likely arises due to an
increase in the rate of differentiation of immature
oligodendrocytes to mature oligodendrocytes rather than an increase
in the rate of differentiation of OPCs to immature
oligodendrocytes.
Example 6
Anti-LINGO-1 Antibody-Mediated Downrequlation of Cell Surface
LINGO-1
[0176] The binding of multi-valent antibodies to cell surface
targets can lead to the internalisation of the antibody:target
complex and subsequent degradation of the target within the
endocytic pathway (Weinmann et al. (2006) Mol Cell Neurosci 32,
161-173).
[0177] To determine the effect of anti-LINGO-1 antibodies on the
amount of cell surface LINGO-1, untransfected CHO-K1 or
CHO-K1-hLINGO-1 cells (see Example 1) are incubated at 37.degree.
C. for 24 hrs with 100 nM 4784, 4785 or 3207 and cell surface
LINGO-1 is subsequently detected with an anti-V5 antibody followed
by an anti-mouse IgG (Fc specific)-POD conjugate developed with a
1-Step.TM. Turbo TMB-ELISA kit (Pierce) (FIG. 6A).
[0178] The amount of cell surface LINGO-1 in CHO-K1-hLINGO-1 cells
is significantly reduced following a 24 hr incubation with
anti-LINGO-1 antibodies 4784 and 4785, whereas incubation with the
control IgG4 3207 has no effect. In addition, incubation with 4785
reduces cell surface LINGO-1 to a greater extent than 4784.
[0179] To assess the effect of anti-LINGO-1 antibodies on the
degradation of cell surface LINGO-1, cell surface proteins on
untransfected CHO-K1 or CHO-K1-hLINGO-1 cells are biotinylated at
4.degree. C. as described (Walmsley et al. (2004) J Cell Sci 117,
4591-4602) and the cells incubated at 37.degree. C. for various
times over a 180 min period with or without 100 nM 4784, 4785 or
3207 (FIG. 6B). At the end of the incubation period, LINGO-1 is
immunoprecipitated from the cell lysate using anti-V5 antibody
coupled to agarose beads and biotinylated LINGO-1 detected in the
precipitate by Western blot analysis using an anti-biotin antibody
(Sigma).
[0180] The intensity of the band corresponding to biotinylated (and
hence cell surface) LINGO-1 diminishes more rapidly in
CHO-K1-hLINGO-1 cells incubated with the anti-LINGO-1 antibodies
4784 and 4785 than in cells incubated without antibody or with the
control IgG4 3207. In addition, incubation with 4785 increases the
rate of degradation of cell surface LINGO-1 to a greater extent
than 4784.
[0181] These results cumulatively show that anti-LINGO-1 antibodies
4784 and 4785 significantly downregulate LINGO-1 at the cell
surface most likely by augmenting the internalisation and
degradation of the protein. This property is expected to contribute
to the efficacy of these antibodies in blocking LINGO-1
function.
Example 7
Enzyme Linked Immunosorbent Assay (ELISA) and FACS Techniques
[0182] Human recombinant LINGO-1-Fc fusion protein is immobilized
onto Maxisorp plates 96 or 384 well for 1 h at RT indirectly by
capturing of the Fc part via a directly immobilized goat anti-human
IgG Fc antibody (100 .mu.l or 20 .mu.l coated at 10 .mu.g/ml in
PBS).
[0183] After coating of 20 .mu.l of the antigen at 5 .mu.g/ml in
PBS, the wells are blocked with PBS/0.05% Tween (PBS-T)/5% milk
powder for 1 h at RT. After washing the wells with PBS-T
BEL-extracts, purified Fabs or control IgGs are diluted in PBS,
added to the wells and incubated for 1 h at RT. To detect the
primary antibodies, the following secondary antibodies are applied:
alkaline phospatase (AP)-conjugated AffiniPure goat F(ab').sub.2
fragment anti-human IgG or anti-mouse IgG (Jackson ImmunoResearch).
For the detection of AP-conjugates fluorogenic substrates like
AttoPhos (Roche) are used according to the manufacturers'
instructions. Between all incubation steps, the wells of the
microtiter plate are washed with PBS-T five times and five times
after the final incubation with secondary antibody. Fluorescence is
measured in a TECAN Spectrafluor plate reader.
FACS Analysis of Antibody Binding to LINGO-1 Expressed on the Cell
Surface of Transfected CHO-K1 Cells
[0184] All stainings are performed in round bottom 96-well
microtiter plates (NUNC.TM., Wiesbaden, Germany) with
2.times.10.sup.5 cells per well. Cells of the respective cell line
are resuspended in PBS/3% FCS/0.02% NaN.sub.3 (FACS buffer) and
mixed with a) antibody from periplasmic extracts or BEL lysates or
b) purified Fab fragments or c) purified IgG diluted in FACS buffer
and incubated at 4.degree. C. for 30-60 min. Cells are then washed
once with 150 .mu.l FACS buffer/well and taken up in 100 .mu.l
phycoerythrin-labeled secondary antibody (R-PE conjugated goat
anti-human IgG (H+L) (Jackson ImmunoResearch) which has been
diluted 1:200 in FACS buffer. After incubation for 30-60 min at
4.degree. C. cells are washed once with FACS buffer, resuspended in
100 .mu.l FACS buffer and binding of LINGO-1 specific antibodies is
measured via FL2 fluorescence intensity of cells in FACSCalibur.TM.
or FACSArray.TM. (Becton Dickinson). For identification of LINGO-1
specific antibodies, stainings are done in parallel using
CHO-K1-cmLINGO-1 or CHO-K1-rLINGO-1. Untransfected CHO-K1 cells
serve as an additional control. Cynomolgus monkey and rat LINGO-1
expressing cells are chosen for screening as these species
orthologues differ only in a few amino acids from the human LINGO-1
protein. Only those clones are judged as being LINGO-1 specific
which are negative on untransfected CHO-K1 cells and
.gtoreq.5.times. above background on LINGO-1 expressing cell
lines.
[0185] Cross-reactivity to human LINGO-1 and other orthologues
(cynomolgus LINGO-1, rat LINGO-1) and to the human LINGO-2
paralogue is tested sequentially.
[0186] After sequence analysis thirty one (31) unique clones are
identified that show strong binding to cell surface expressed human
LINGO-1 in FACS analysis. Twelve (12) binders show strong binding
to captured human LINGO-1-Fc in ELISA (signal:noise ratio greater
than 10:1) and seven (7) show intermediate binding in ELISA
(signal:noise ratio greater than 5:1). Four (4) of the binders
showed strong binding to captured human NgR-Fc fusion protein
(R&D Systems) in ELISA and are discontinued. Another three (3)
of the binders do not cross-react to all of the three species of
LINGO-1 and are discontinued. The remaining 24 clones that are
cross-reactive to human/cynomolgus monkey/rat LINGO-1 but not to
human NgR-Fc are expressed, purified and tested for their ability
to significantly inhibit the binding of LINGO-1 to NgR (see FIG. 1)
and disinhibit the neurite outgrowth inhibitory activity of spinal
cord myelin in vitro (see FIGS. 2-4) leading to the selection of
Fabs 4784 and 4785 for further analysis. In an ELISA, 4784 and 4785
bind to captured human LINGO-1-Fc but no binding is observed to
human LINGO-1-.DELTA.LRR-Fc or human NgR-Fc compared to an
unrelated Fc control (see Table 1 and FIG. 7). This indicates that
4784 and 4785 have epitopes that are within the LRR region
(residues 66-353) of LINGO-1.
TABLE-US-00002 TABLE 1 Characterization of anti-LINGO-1 Fabs by
ELISA Human Human LINGO-1 Human LINGO- Human Unrelated LINGO-1 Fc
1.DELTA.LRR-Fc NgR-Fc Fc 4784 98 49 68 52 4785 113 8 7 6 Values for
ELISA analyses are given as mean values of relative fluorescence
units.
Affinity Determination of Selected Anti-LINGO-1 Fabs Using FACS
Saturation Analysis
[0187] Cell based affinity of anti-LINGO-1 specific antibodies is
determined by FACS saturation binding experiments. As the
concentration of the antigen present in the sample to stain
influences the apparent K.sub.D values, only 1.25.times.10.sup.4
cells/well in contrast to 2.times.10.sup.5 cells/well are used in
order to reduce the antigen concentration in FACS saturation
experiments. Otherwise the staining procedure is done identical to
the FACS staining procedure described above.
[0188] In detail, CHO-K1-hLINGO-1, CHO-K1-cmLINGO-1 or
CHO-K1-rLINGO-1 are detached from culture flasks by versene, washed
with FACS buffer and resuspended in FACS buffer. Purified
anti-LINGO-1 Fabs are serially diluted in FACS buffer and spread
into round bottom 96-well microtiter plates (NUNC.TM., Wiesbaden,
Germany). For each concentration, duplicate wells are incubated
with 1.25.times.10.sup.4 cells for 30-60 min on ice in a total
volume of 100 .mu.l. After a washing step by applying 150 .mu.l
FACS buffer and centrifugation for 5 min at 400.times.g, the cell
pellets are resuspended in 100 .mu.l phycoerythrin-labeled
secondary antibody (R-PE conjugated goat anti-human IgG (H+L)
(Jackson ImmunoResearch) which has been diluted 1:200 in FACS
buffer. After incubation for 30-60 min at 4.degree. C. cells are
washed once with FACS buffer, resuspended in 100 .mu.l FACS buffer
and binding of LINGO-1 specific antibodies is measured via FL2
fluorescence intensity of cells in FACSArray.TM. (Becton
Dickinson). Apparent K.sub.D values/EC50 values are determined from
the saturation binding curves using GraphPad Prism v3.03 software
or GraphPad Prism v4.03 applying a non-linear regression curve
fit.
[0189] Using this assay the following apparent K.sub.D values can
be determined (Table 2). In Fab format the clone 4784 has rather
weak affinities to human LINGO-1, cynomolgus monkey LINGO-1 and rat
LINGO-1 (14.07 nM, 27.11, and 24.03 nM respectively). However,
clone 4784 does not bind to human LINGO-2 in the Fab format. In Fab
format the clone 4785 shows subnanomolar binding affinities (i.e.
apparent K.sub.D values being less than 1.times.10.sup.-9 M) to
human LINGO-1, cynomolgus monkey LINGO-1 and rat LINGO-1. Clone
4785 shows cross-reactivity to human LINGO-2 in Fab format with low
nanomolar to subnanomolar affinity. The consequence of
cross-reactivity to LINGO-2 cannot be assessed at the time of
writing as LINGO-2 function and distribution are as yet unknown.
However, beneficial effects cannot be excluded.
TABLE-US-00003 TABLE 2 Apparent K.sub.D values of anti-LINGO-1 Fabs
to LINGO-1 or LINGO-2 expressed by CHO-K1 cells Human Human
Cynomolgus Rat LINGO-1 LINGO-2 LINGO-1 LINGO-1 4784 14.07 nb 27.11
24.03 4785 0.35 1.21 0.26 0.260 Values given are mean values of
apparent K.sub.Ds in nM. nb, not binding.
Example 8
Cloning, Expression and Purification of HuCAL.RTM. IgG4
[0190] Conversion into the IgG Format
[0191] In order to express full length immunoglobulin (Ig),
variable domain fragments of heavy (VH) and light chains (VL) are
subcloned from the pMORPH.RTM.X9_MH (SEQ ID NO: 39) Fab expression
vectors either into the pMORPH.RTM._h_Ig (SEQ ID NOS: 40-42) or the
pMORPH.RTM.2_h_Ig (SEQ ID NOS: 43-45) vector series for human
IgG4.
[0192] Restriction enzymes EcoRI, MfeI, and BlpI are used for
subcloning of the VH domain fragment into pMORPH.RTM._h_IgG4 (SEQ
ID NO: 40): the vector backbone is generated by EcoRI/BlpI
digestion and extraction of the 6400 bp fragment whereas the VH
fragment (350 bp) is produced by digestion with MfeI and BlpI and
subsequent purification. Vector and insert are ligated via
compatible overhangs generated by the EcoRI and MfeI digests,
respectively, and via the BlpI site. Thereby, both the EcoRI and
the MfeI restriction site are destroyed.
[0193] Restriction enzymes MfeI and BlpI are used for subcloning of
the VH domain fragment into pMORPH.RTM.2_h_IgG4 (SEQ ID NO: 43). In
this new generation of IgG vectors, upon other modifications, the
EcoRI site (which allowed only sub-cloning via compatible
overhangs) is replaced by the MfeI site thus allowing MfeI/BlpI
digestion of both, vector and insert.
[0194] Subcloning of the VL domain fragment into
pMORPH.RTM._h_Ig.kappa. (SEQ ID NO: 42) and
pMORPH.RTM.2_h_Ig.kappa. (SEQ ID NO: 45) is performed via the EcoRV
and BsiWI sites, whereas subcloning into pMORPH.RTM._h_Ig.lamda.
(SEQ ID NO: 41) and pMORPH.RTM.2_h_Ig.lamda.2 (SEQ ID NO: 43) is
done using EcoRV and HpaI.
Transient Expression and Purification of Human IgG
[0195] HEK293 cells are transfected with an equimolar amount of IgG
heavy and light chain expression vectors. On days 4 or 5
post-transfection the cell culture supernatant is harvested. After
adjusting the pH of the supernatant to 8.0 and sterile filtration,
the solution is subjected to standard protein A column
chromatography (Poros 20A, PE Biosystems).
Example 9
Affinity Determination of Selected Anti-LINGO-1 IgG4s Using FACS
Saturation Analysis
[0196] Cell based affinity of anti-LINGO-1 specific antibodies is
determined by FACS saturation binding experiments. The
determination of the apparent K.sub.D values is carried out
identical to the procedure described above using anti-LINGO-1 Fab
antibodies.
[0197] In detail, CHO-K1-hLINGO-1, CHO-K1-cmLINGO-1 or
CHO-K1-rLINGO-1 are detached from culture flasks by versene, washed
with FACS buffer and resuspended in FACS buffer. Purified
anti-LINGO-1 IgG4s are serially diluted in FACS buffer and spread
into round bottom 96-well microtiter plates (NUNC.TM., Wiesbaden,
Germany). For each concentration, duplicate wells are incubated
with 1.25.times.10.sup.4 cells for 30-60 min on ice in a total
volume of 100 .mu.l. After a washing step by applying 150 .mu.l
FACS buffer and centrifugation for 5 min at 400.times.g, the cell
pellets are resuspended in 10 .mu.l phycoerythrin-labeled secondary
antibody (R-PE conjugated goat anti-human IgG (H+L) (Jackson
ImmunoResearch) which has been diluted 1:200 in FACS buffer. After
incubation for 30-60 min at 4.degree. C. cells are washed once with
FACS buffer, resuspended in 100 .mu.l FACS buffer and binding of
LINGO-1 specific antibodies is measured via FL2 fluorescence
intensity of cells in FACSArray.TM. (Becton Dickinson). Apparent
K.sub.D values/EC50 values are determined from the saturation
binding curves using GraphPad Prism v3.03 software or GraphPad
Prism v4.03 applying a non-linear regression curve fit. Using this
assay the following apparent K.sub.D values can be determined
(Table 3).
[0198] The affinity of 4784 and 4785 IgG4 antibodies produced by
using the pMORPH.RTM.2_h_Ig vector series are shown in Table 3.
4784 and 4785 in the IgG4 format have apparent K.sub.D values
clearly below 1 nM to human, cynomolgus and rat LINGO-1. 4784 has a
far lower cross-reactivity to human LINGO-2 than 4785.
TABLE-US-00004 TABLE 3 Apparent K.sub.D values of anti-LINGO-1
IgG4s to LINGO-1 or LINGO-2 expressed by CHO-K1 cells Human Human
Cynomolgus Rat LINGO-1 LINGO-2 LINGO-1 LINGO-1 4784 0.29 25.94 0.62
0.98 4785 0.07 0.95 0.18 0.07 Values given are mean values of
apparent K.sub.Ds in nM.
Example 10
Influence of Human Cerebro-Spinal Fluid on Binding of Selected
Anti-LINGO-1 IgG4s to Human LINGO-1 Using FACS Analysis
[0199] Influence of human cerebro-spinal fluid on binding of
anti-LINGO-1 IgG4s to human LINGO-1 is tested by FACS saturation
binding experiments. Serial dilutions of the 4784 and 4785 are
prepared. Binding to CHO-K1-hLINGO-1 is tested in the presence of
50% human cerebro-spinal fluid. The cells are stained in the
presence of human CSF with these IgG4 antibodies according to the
FACS stainings described above.
[0200] In detail, CHO-K1-hLINGO-1 are detached from culture flasks
by versene, washed with FACS buffer and resuspended in FACS buffer.
Purified anti-LINGO-1 IgG4s are serially diluted in FACS buffer
plus 50% human serum and incubated for 60 min at 4.degree. C. As
controls, serial dilutions of the candidate binders in IgG4 format
are incubated in FACS buffer with 2.6% BSA resembling protein
content of human cerebro-spinal fluid for 60 min at 4.degree. C.
After incubation the serial dilutions are spread into round bottom
96-well microtiter plates (NUNC.TM., Wiesbaden, Germany). For each
concentration, duplicate wells are incubated with
1.25.times.10.sup.4 cells for 30-60 min on ice in a total volume of
100 .mu.l. After three washing steps by applying 150 .mu.l FACS
buffer and centrifugation for 5 min at 400.times.g, the cell
pellets are resuspended in 100 .mu.l phycoerythrin-labeled
secondary antibody (R-PE conjugated goat anti-human IgG (H+L)
(Jackson ImmunoResearch) which has been diluted 1:200 in FACS
buffer. After incubation for 30-60 min at 4.degree. C. cells are
washed once with FACS buffer, resuspended in 100 .mu.l FACS buffer
and binding of LINGO-1 specific antibodies is measured via FL2
fluorescence intensity of cells in FACSArray.TM. (Becton
Dickinson). Apparent K.sub.D values/EC50 values are determined from
the saturation binding curves using GraphPad Prism v3.03 software
or GraphPad Prism v4.03 applying a non-linear regression curve
fit.
[0201] Using this assay the influence of 50% human cerebrospinal
fluid could be compared to the controls (Table 4). Incubation in
50% human cerebro-spinal fluid leads to a decrease in binding
affinity with all binders being affected differently. The strongest
impact on binding affinity by the presence of human cerebro-spinal
fluid is seen for 4784 which shows a reduction in affinity by 73%
from 0.43 nM to 1.57 nM.
TABLE-US-00005 TABLE 4 Influence of Human Cerebro-spinal Fluid on
Apparent K.sub.D values of anti-LINGO-1 IgG4s to LINGO-1 expressed
by CHO-K1 cells App. K.sub.D w/o 50% App. K.sub.D ratio w/o CSF
App. K.sub.D w/ CSF CSF:w/ CSF 4784 0.43 1.57 0.27 4785 0.19 0.25
0.76 Values given are mean values of apparent K.sub.Ds in nM.
Example 11
Influence of Human Serum on Binding of Selected Anti-LINGO-1 IgG4s
to Human LINGO-1 Using FACS Analysis
[0202] Influence of human serum on binding of anti-LINGO-1 IgG4s to
human LINGO-1 is tested by FACS saturation binding experiments.
Serial dilutions of 4784 and 4785 are prepared in the presence of
50% v/v human serum. After incubation for 60 min cells are stained
with these preincubated IgG4 antibodies according to the FACS
stainings described above.
[0203] In detail, CHO-K1-hLINGO-1 are detached from culture flasks
by versene, washed with FACS buffer and resuspended in FACS buffer.
Purified anti-LINGO-1 IgG4s are serially diluted in FACS buffer
plus 50% human serum and incubated for 60 min at 4.degree. C. As
controls, serial dilutions of the candidate binders in IgG4 format
are incubated in FACS buffer plus 2.6% BSA resembling protein
content of human serum or are incubated in FACS buffer alone for 60
min at 4.degree. C. After incubation the serial dilutions are
spread into round bottom 96-well microtiter plates (NUNC.TM.,
Wiesbaden, Germany). For each concentration, duplicate wells are
incubated with 1.25.times.10.sup.4 cells for 30-60 min on ice in a
total volume of 100 .mu.l. After three washing steps by applying
150 .mu.l FACS buffer and centrifugation for 5 min at 400.times.g,
the cell pellets are resuspended in 100 .mu.l phycoerythrin-labeled
secondary antibody (R-PE conjugated goat anti-human IgG (H+L)
(Jackson ImmunoResearch) which has been diluted 1:200 in FACS
buffer. After incubation for 30-60 min at 4.degree. C. cells are
washed once with FACS buffer, resuspended in 100 .mu.l FACS buffer
and binding of LINGO-1 specific antibodies is measured via FL2
fluorescence intensity of cells in FACSArray.TM. (Becton
Dickinson). Apparent K.sub.D values/EC50 values are determined from
the saturation binding curves using GraphPad Prism v3.03 software
or GraphPad Prism v4.03 applying a non-linear regression curve
fit.
[0204] Using this assay the influence of preincubation in 50% human
serum can be compared to the controls (Table 5). Incubation for 1
hr in the presence of human serum has no effect on the K.sub.D
values of 4784 and 4785. These antibodies are therefore stable in
human serum over this time period and, furthermore, as their
K.sub.D s are unchanged, they do not appear to cross-react with
serum components.
TABLE-US-00006 TABLE 5 Influence of Human Serum on Apparent K.sub.D
values of anti-LINGO-1 IgG4s to LINGO-1 expressed by CHO-K1 cells
FACS FB + 2.6% FB + Buffer (FB) BSA 50% HS 4784 0.28 0.19 0.27 4785
0.08 0.05 0.06 Values given are mean values of apparent K.sub.Ds in
nM.
TABLE-US-00007 List of Sequences with short description SEQ ID NO:
1 Rat mature LINGO-1 ectodomain (residues 34-550)
TGCPPRCECSAQDRAVLCHRKRFVAVPEGIPTETRLLDLGKNRIKTLNQDEFASFPHLEE
LELNENIVSAVEPGAFNNLFNLRTLGLRSNRLKLIPLGVFTGLSNLTKLDISENKIVILL
DYMFQDLYNLKSLEVGDNDLVYISHRAFSGLNSLEQLTLEKCNLTSIPTEALSHLHGLIV
LRLRHLNINAIRDYSFKRLYRLKVLEISHWPYLDTMTPNCLYGLNLTSLSITHCNLTAVP
YLAVRHLVYLRFLNLSYNPIGTIEGSMLHELLRLQEIQLVGGQLAVVEPYAFRGLNYLRV
LNVSGNQLTTLEESAFHSVGNLETLILDSNPLACDCRLLWVFRRRWRLNFNRQQPTCATP
EFVQGKEFKDFPDVLLPNYFTCRRAHIRDRKAQQVFVDEGHTVQFVCRADGDPPPAILWL
SPRKHLVSAKSNGRLTVFPDGTLEVRYAQVQDNGTYLCIAANAGGNDSMPAHLHVRSYSP
DWPHQPNKTFAFISNQPGEGEANSTRATVPFPFDIKT SEQ ID NO: 2 Cynomologus
mature LINGO-1 ectodomain (residues 34-550)
TGCPPRCECSAQDRAVLCHRKRFVAVPEGIPTETRLLDLGKNRIKTLNQDEFASFPHLEE
LELNENIVSAVEPGAFNNLFNLRTLGLRSNRLKLIPLGVFTGLSNLTKLDISENKIVILL
DYMFQDLYNLKSLEVGDNDLVYISHRAFSGLNSLEQLTLEKCNLTSIPTEALSHLHGLIV
LRLRHLNINAIRDYSFKRLYRLKVLEISHWPYLDTMTPNCLYGLNLTSLSITHCNLTAVP
YLAVRHLVYLRFLNLSYNPISTIEGSMLHELLRLQEIQLVGGQLAMVEPYAFRGLNYLRV
LNVSGNQLTTLEESVFHSVGNLETLILDSNPLACDCRLLWVFRRRWRLNFNRQQPTCATP
EFVQGKEFKDFPDVLLPNYFTCRRARIRDRKAQQVFVDEGHTVQFVCRADGDPPPAILWL
SPRKHLVSAKSNGRLTVFPDGTLEVRYAQVQDNGTYLCIAANAGGNDSMPAHLHVRSYSP
DWPHQPNKTFAFIPNQPGEGEANSTRATVPFPFDIKT SEQ ID NO: 3 Human mature
LINGO-1 ectodomain (residues 34-550)
TGCPPRCECSAQDRAVLCHRKRFVAVPEGIPTETRLLDLGKNRIKTLNQDEFASFPHLEE
LELNENIVSAVEPGAFNNLFNLRTLGLRSNRLKLIPLGVFTGLSNLTKLDISENKIVILL
DYMFQDLYNLKSLEVGDNDLVYISHRAFSGLNSLEQLTLEKCNLTSIPTEALSHLHGLIV
LRLRHLNINAIRDYSFKRLYRLKVLEISHWPYLDTMTPNCLYGLNLTSLSITHCNLTAVP
YLAVRHLVYLRFLNLSYNPISTIEGSMLHELLRLQEIQLVGGQLAVVEPYAFRGLNYLRV
LNVSGNQLTTLEESVFHSVGNLETLILDSNPLACDCRLLWVFRRRWRLNFNRQQPTCATP
EFVQGKEFKDFPDVLLPNYFTCRRARIRDRKAQQVFVDEGHTVQFVCRADGDPPPAILWL
SPRKHLVSAKSNGRLTVFPDGTLEVRYAQVQDNGTYLCIAANAGGNDSMPAHLHVRSYSP
DWPHQPNKTFAFISNQPGEGEANSTRATVPFPFDIKT SEQ ID NO: 4 4784 V.sub.L
DIELTQPPSVSVAPGQTARISCSGDNIGNYYVYWYQQKPGQAPVLVIYEDTNRPSGIPERFSGSNSGNTATLTI-
S GTQAEDEADYYCQSYDNLHEQVFGGGTKLTVLG SEQ ID NO: 5 4784 V.sub.H
QVQLKESGPALVKPTQTLTLTCTFSGFSLSSSGVGVGWIRQPPGKALEWLAHIGSDDDKYYSTSLKTRLTISKD-
T SKNQVVLTMTNMDPVDTATYYCARNQQYGDGYPGYFDYWGQGTLVTVSS SEQ ID NO: 6
4785 V.sub.L
DIVLTQPPSVSGAPGQRVTISCSGSSSNIGNNYVSWYQQLPGTAPKLLISRNSKRPSGVPDRFSGSKSGTSASL-
A ITGLQSEDEADYYCSTYDTFSIVFGGGTKLTVLG SEQ ID NO: 7 4785 V.sub.H
QVQLQQSGPGLVKPSQTLSLTCAISGDSVSDNSAAWSWIRQSPGRGLEWLGLIYLRSKWDNDYAVSVKSRITIN-
P DTSKNQFSLQLNSVTPEDTAVYYCARTGRADEFDVWGQGTLVTVSS SEQ ID NO: 8
DNA-4784 V.sub.H
CAGGTGCAATTGAAAGAAAGCGGCCCGGCCCTGGTGAAACCGACCCAAACCCTGACCCTGACCTGTACCTTTTC-
C
GGATTTAGCCTGTCTTCTTCTGGTGTTGGTGTGGGTTGGATTCGCCAGCCGCCTGGGAAAGCCCTCGAGTGGCT-
G
GCTCATATCGGTTCTGATGATGATAAGTATTATAGCACCAGCCTGAAAACGCGTCTGACCATTAGCAAAGATAC-
T
TCGAAAAATCAGGTGGTGCTGACTATGACCAACATGGACCCGGTGGATACGGCCACCTATTATTGCGCGCGTAA-
T
CAGCAGTATGGTGATGGTTATCCTGGTTATTTTGATTATTGGGGCCAAGGCACCCTGGTGACGGTTAGCTCA
SEQ ID NO: 9 DNA-4785 V.sub.H
CAGGTGCAATTGCAACAGTCTGGTCCGGGCCTGGTGAAACCGAGCCAAACCCTGAGCCTGACCTGTGCGATTTC-
C
GGAGATAGCGTGAGCGATAATTCTGCTGCTTGGTCTTGGATTCGCCAGTCTCCTGGGCGTGGCCTCGAGTGGCT-
G
GGCCTTATCTATCTTCGTAGCAAGTGGGATAACGATTATGCGGTGAGCGTGAAAAGCCGGATTACCATCAACCC-
G
GATACTTCGAAAAACCAGTTTAGCCTGCAACTGAACAGCGTGACCCCGGAAGATACGGCCGTGTATTATTGCGC-
G CGTACTGGTCGTGCTGATGAGTTTGATGTTTGGGGCCAAGGCACCCTGGTGACGGTTAGCTCA
SEQ ID NO: 10 DNA-4784 V.sub.L
GATATCGAACTGACCCAGCCGCCTTCAGTGAGCGTTGCACCAGGTCAGACCGCGCGTATCTCGTGTAGCGGCGA-
T
AATATTGGTAATTATTATGTTTATTGGTACCAGCAGAAACCCGGGCAGGCGCCAGTTCTTGTGATTTATGAGGA-
T
ACTAATCGTCCCTCAGGCATCCCGGAACGCTTTAGCGGATCCAACAGCGGCAACACCGCGACCCTGACCATTAG-
C
GGCACTCAGGCGGAAGACGAAGCGGATTATTATTGCCAGTCTTATGATAATCTTCATGAGCAGGTGTTTGGCGG-
C GGCACGAAGTTAACCGTTCTTGGCCAG SEQ ID NO: 11 DNA-4785 V.sub.L
GATATCGTGCTGACCCAGCCGCCTTCAGTGAGTGGCGCACCAGGTCAGCGTGTGACCATCTCGTGTAGCGGCAG-
C
AGCAGCAACATTGGTAATAATTATGTGTCTTGGTACCAGCAGTTGCCCGGGACGGCGCCGAAACTTCTGATTTC-
T
CGTAATTCTAAGCGTCCCTCAGGCGTGCCGGATCGTTTTAGCGGATCCAAAAGCGGCACCAGCGCGAGCCTTGC-
G
ATTACGGGCCTGCAAAGCGAAGACGAAGCGGATTATTATTGCTCTACTTATGATACTTTTTCTATTGTGTTTGG-
C GGCGGCACGAAGTTAACCGTTCTTGGCCAG SEQ ID NO: 12 Antibody 4784 CDR-H1
SSGVGVG SEQ ID NO: 13 Antibody 4784 CDR-H2 HIGSDDDKYYSTSLKT SEQ ID
NO: 14 Antibody 4784 CDR-H3 NQQYGDGYPGYFDY SEQ ID NO: 15 Antibody
4784 CDR-L1 SGDNIGNYYVY SEQ ID NO: 16 Antibody 4784 CDR-L2 EDTNRPS
SEQ ID NO: 17 Antibody 4784 CDR-L3 QSYDNLHEQV SEQ ID NO: 18
Antibody 4785 CDR'-H1 DNSAAWS SEQ ID NO: 19 Antibody 4785 CDR'-H2
LIYLRSKWDNDYAVSVKS SEQ ID NO: 20 Antibody 4785 CDR'-H3 TGRADEFDV
SEQ ID NO: 21 Antibody 4785 CDR'-L1 SGSSSNIGNNYVS SEQ ID NO: 22
Antibody 4785 CDR'-L2 RNSKRPS SEQ ID NO: 23 Antibody 4785 CDR'-L3
STYDTFSIV SEQ ID NO: 24 Forward primer DM22
GGTTATCTCGAGACCGGCTGCCCGCCCC SEQ ID NO: 25 Reverse primer DM23
GGCCCTTCTAGATCACTCGCCTGGCTGGTTGGAGATG SEQ ID NO: 26 APtag-5-NHIS
vector
gacggatcgggagatctcccgatcccctatggtcgactctcagtacaatctgctctgatgccgcatagttaagc-
c
agtatctgctccctgcttgtgtgttggaggtcgctgagtagtgcgcgagcaaaatttaagctacaacaaggcaa-
g
gcttgaccgacaattgcatgaagaatctgcttagggttaggcgttttgcgctgcttcgcgatgtacgggccaga-
t
atacgcgttgacattgattattgactagttattaatagtaatcaattacggggtcattagttcatagcccatat-
a
tggagttccgcgttacataacttacggtaaatggcccgcctggctgaccgcccaacgacccccgcccattgacg-
t
caataatgacgtatgttcccatagtaacgccaatagggactttccattgacgtcaatgggtggactatttacgg-
t
aaactgcccacttggcagtacatcaagtgtatcatatgccaagtacgccccctattgacgtcaatgacggtaaa-
t
ggcccgcctggcattatgcccagtacatgaccttatgggactttcctacttggcagtacatctacgtattagtc-
a
tcgctattaccatggtgatgcggttttggcagtacatcaatgggcgtggatagcggtttgactcacggggattt-
c
caagtctccaccccattgacgtcaatgggagtttgttttggcaccaaaatcaacgggactttccaaaatgtcgt-
a
acaactccgccccattgacgcaaatgggcggtaggcgtgtacggtgggaggtctatataagcagagctctctgg-
c
taactagagaacccactgcttactggcttatcgaaattaatacgactcactatagggagacccaagctggctag-
c
caccatggagacagacacactcctgctatgggtactgctgctctgggttccaggttccactggtgacgcggccc-
a
gccggcccatcatcatcatcatcatgaagcttacgtaagatcttccggaatcatcccagttgaggaggagaacc-
c
ggacttctggaaccgcgaggcagccgaggccctgggtgccgccaagaagctgcagcctgcacagacagccgcca-
a
gaacctcatcatcttcctgggcgatgggatgggggtgtctacggtgacagctgccaggatcctaaaagggcaga-
a
gaaggacaaactggggcctgagatacccctggccatggaccgcttcccatatgtggctctgtccaagacataca-
a
tgtagacaaacatgtgccagacagtggagccacagccacggcctacctgtgcggggtcaagggcaacttccaga-
c
cattggcttgagtgcagccgcccgctttaaccagtgcaacacgacacgcggcaacgaggtcatctccgtgatga-
a
tcgggccaagaaagcagggaagtcagtgggagtggtaaccaccacacgagtgcagcacgcctcgccagccggca-
c
ctacgcccacacggtgaaccgcaactggtactcggacgccgacgtgcctgcctcggcccgccaggaggggtgcc-
a
ggacatcgctacgcagctcatctccaacatggacattgacgtgatcctaggtggaggccgaaagtacatgtttc-
g
catgggaaccccagaccctgagtacccagatgactacagccaaggtgggaccaggctggacgggaagaatctgg-
t
gcaggaatggctggcgaagcgccagggtgcccggtatgtgtggaaccgcactgagctcatgcaggcttccctgg-
a
cccgtctgtgacccatctcatgggtctctttgagcctggagacatgaaatacgagatccaccgagactccacac-
t
ggacccctccctgatggagatgacagaggctgccctgcgcctgctgagcaggaacccccgcggcttcttcctct-
t
cgtggagggtggtcgcatcgaccatggtcatcatgaaagcagggcttaccgggcactgactgagacgatcatgt-
t
cgacgacgccattgagagggcgggccagctcaccagcgaggaggacacgctgagcctcgtcactgccgaccact-
c
ccacgtcttctccttcggaggctaccccctgcgagggagctccatcttcgggctggcccctggcaaggcccggg-
a
caggaaggcctacacggtcctcctatacggaaacggtccaggctatgtgctcaaggacggcgcccggccggatg-
t
taccgagagcgagagcgggagccccgagtatcggcagcagtcagcagtgcccctggacgaagagacccacgcag-
g
cgaggacgtggcggtgttcgcgcgcggcccgcaggcgcacctggttcacggcgtgcaggagcagaccttcatag-
c
gcacgtcatggccttcgccgcctgcctggagccctacaccgcctgcgacctggcgccccccgccggcaccaccg-
a
cgcgcgcacccgggttatctcgaggaagcgctctctctagaagggcccgaacaaaaactcatctcagaagagga
tctgaatagcgccgtcgaccatcatcatcatcatcattgagtttaaacccgctgatcagcctcgactgtgcctt-
c
tagttgccagccatctgttgtttgcccctcccccgtgccttccttgaccctggaaggtgccactcccactgtcc-
t
ttcctaataaaatgaggaaattgcatcgcattgtctgagtaggtgtcattctattctggggggtggggtggggc-
a
ggacagcaagggggaggattgggaagacaatagcaggcatgctggggatgcggtgggctctatggcttctgagg-
c
ggaaagaaccagctggggctctagggggtatccccacgcgccctgtagcggcgcattaagcgcggcgggtgtgg-
t
ggttacgcgcagcgtgaccgctacacttgccagcgccctagcgcccgctcctttcgctttcttcccttcctttc-
t
cgccacgttcgccggctttccccgtcaagctctaaatcggggcatccctttagggttccgatttagtgctttac-
g
gcacctcgaccccaaaaaacttgattagggtgatggttcacgtagtgggccatcgccctgatagacggtttttc-
g
ccctttgacgttggagtccacgttctttaatagtggactcttgttccaaactggaacaacactcaaccctatct-
c
ggtctattcttttgatttataagggattttggggatttcggcctattggttaaaaaatgagctgatttaacaaa-
a
atttaacgcgaattaattctgtggaatgtgtgtcagttagggtgtggaaagtccccaggctccccagcaggcag-
a
agtatgcaaagcatgcatctcaattagtcagcaaccaggtgtggaaagtccccaggctccccagcaggcagaag-
t
atgcaaagcatgcatctcaattagtcagcaaccatagtcccgcccctaactccgcccatcccgcccctaactcc-
g
cccagttccgcccattctccgccccatggctgactaattttttttatttatgcagaggccgaggccgcctctgc-
c
tctgagctattccagaagtagtgaggaggcttttttggaggcctaggcttttgcaaaaagctcccgggagcttg-
t
atatccattttcggatctgatcagcacgtgttgacaattaatcatcggcatagtatatcggcatagtataatac-
g
acaaggtgaggaactaaaccatggccaagttgaccagtgccgttccggtgctcaccgcgcgcgacgtcgccgga-
g
cggtcgagttctggaccgaccggctcgggttctcccgggacttcgtggaggacgacttcgccggtgtggtccgg-
g
acgacgtgaccctgttcatcagcgcggtccaggaccaggtggtgccggacaacaccctggcctgggtgtgggtg-
c
gcggcctggacgagctgtacgccgagtggtcggaggtcgtgtccacgaacttccgggacgcctccgggccggcc-
a
tgaccgagatcggcgagcagccgtgggggcgggagttcgccctgcgcgacccggccggcaactgcgtgcacttc-
g
tggccgaggagcaggactgacacgtgctacgagatttcgattccaccgccgccttctatgaaaggttgggcttc-
g
gaatcgttttccgggacgccggctggatgatcctccagcgcggggatctcatgctggagttcttcgcccacccc-
a
acttgtttattgcagcttataatggttacaaataaagcaatagcatcacaaatttcacaaataaagcatttttt-
t
cactgcattctagttgtggtttgtccaaactcatcaatgtatcttatcatgtctgtataccgtcgacctctagc-
t
agagcttggcgtaatcatggtcatagctgtttcctgtgtgaaattgttatccgctcacaattccacacaacata-
c
gagccggaagcataaagtgtaaagcctggggtgcctaatgagtgagctaactcacattaattgcgttgcgctca-
c
tgcccgctttccagtcgggaaacctgtcgtgccagctgcattaatgaatcggccaacgcgcggggagaggcggt-
t
tgcgtattgggcgctcttccgcttcctcgctcactgactcgctgcgctcggtcgttcggctgcggcgagcggta-
t
cagctcactcaaaggcggtaatacggttatccacagaatcaggggataacgcaggaaagaacatgtgagcaaaa-
g
gccagcaaaaggccaggaaccgtaaaaaggccgcgttgctggcgtttttccataggctccgcccccctgacgag-
c
atcacaaaaatcgacgctcaagtcagaggtggcgaaacccgacaggactataaagataccaggcgtttccccct-
g
gaagctccctcgtgcgctctcctgttccgaccctgccgcttaccggatacctgtccgcctttctcccttcggga-
a
gcgtggcgctttctcaatgctcacgctgtaggtatctcagttcggtgtaggtcgttcgctccaagctgggctgt-
g
tgcacgaaccccccgttcagcccgaccgctgcgccttatccggtaactatcgtcttgagtccaacccggtaaga-
c
acgacttatcgccactggcagcagccactggtaacaggattagcagagcgaggtatgtaggcggtgctacagag-
t
tcttgaagtggtggcctaactacggctacactagaaggacagtatttggtatctgcgctctgctgaagccagtt-
a
ccttcggaaaaagagttggtagctcttgatccggcaaacaaaccaccgctggtagcggtggtttttttgtttgc-
a
agcagcagattacgcgcagaaaaaaaggatctcaagaagatcctttgatcttttctacggggtctgacgctcag-
t
ggaacgaaaactcacgttaagggattttggtcatgagattatcaaaaaggatcttcacctagatccttttaaat-
t
aaaaatgaagttttaaatcaatctaaagtatatatgagtaaacttggtctgacagttaccaatgcttaatcagt-
g
aggcacctatctcagcgatctgtctatttcgttcatccatagttgcctgactccccgtcgtgtagataactacg-
a
tacgggagggcttaccatctggccccagtgctgcaatgataccgcgagacccacgctcaccggctccagattta-
t
cagcaataaaccagccagccggaagggccgagcgcagaagtggtcctgcaactttatccgcctccatccagtct-
a
ttaattgttgccgggaagctagagtaagtagttcgccagttaatagtttgcgcaacgttgttgccattgctaca-
g
gcatcgtggtgtcacgctcgtcgtttggtatggcttcattcagctccggttcccaacgatcaaggcgagttaca-
t
gatcccccatgttgtgcaaaaaagcggttagctccttcggtcctccgatcgttgtcagaagtaagttggccgca-
g
tgttatcactcatggttatggcagcactgcataattctcttactgtcatgccatccgtaagatgcttttctgtg-
a
ctggtgagtactcaaccaagtcattctgagaatagtgtatgcggcgaccgagttgctcttgcccggcgtcaata-
c
gggataataccgcgccacatagcagaactttaaaagtgctcatcattggaaaacgttcttcggggcgaaaactc-
t
caaggatcttaccgctgttgagatccagttcgatgtaacccactcgtgcacccaactgatcttcagcatctttt-
a
ctttcaccagcgtttctgggtgagcaaaaacaggaaggcaaaatgccgcaaaaaagggaataagggcgacacgg-
a
aatgttgaatactcatactcttcctttttcaatattattgaagcatttatcagggttattgtctcatgagcgga-
t
acatatttgaatgtatttagaaaaataaacaaataggggttccgcgcacatttccccgaaaagtgccacctgac-
g tc SEQ ID NO: 27 human LINGO-1 mature DNA sequence
acgggctgcccgccccgctgcgagtgctccgcccaggaccgcgctgtgctgtgccaccgcaagcgctttgtggc-
a
gtccccgagggcatccccaccgagacgcgcctgctggacctaggcaagaaccgcatcaaaacgctcaaccagga-
c
gagttcgccagcttcccgcacctggaggagctggagctcaacgagaacatcgtgagcgccgtggagcccggcgc-
c
ttcaacaacctcttcaacctccggacgctgggtctccgcagcaaccgcctgaagctcatcccgctaggcgtctt-
c
actggcctcagcaacctgaccaagctggacatcagcgagaacaagattgttatcctgctggactacatgtttca-
g
gacctgtacaacctcaagtcactggaggttggcgacaatgacctcgtctacatctctcaccgcgccttcagcgg-
c
ctcaacagcctggagcagctgacgctggagaaatgcaacctgacctccatccccaccgaggcgctgtcccacct-
g
cacggcctcatcgtcctgaggctccggcacctcaacatcaatgccatccgggactactccttcaagaggctcta-
c
cgactcaaggtcttggagatctcccactggccctacttggacaccatgacacccaactgcctctacggcctcaa-
c
ctgacgtccctgtccatcacacactgcaatctgaccgctgtgccctacctggccgtccgccacctagtctatct-
c
cgcttcctcaacctctcctacaaccccatcagcaccattgagggctccatgttgcatgagctgctccggctgca-
g
gagatccagctggtgggcgggcagctggccgtggtggagccctatgccttccgcggcctcaactacctgcgcgt-
g
ctcaatgtctctggcaaccagctgaccacactggaggaatcagtcttccactcggtgggcaacctggagacact-
c
atcctggactccaacccgctggcctgcgactgtcggctcctgtgggtgttccggcgccgctggcggctcaactt-
c
aaccggcagcagcccacgtgcgccacgcccgagtttgtccagggcaaggagttcaaggacttccctgatgtgct-
a
ctgcccaactacttcacctgccgccgcgcccgcatccgggaccgcaaggcccagcaggtgtttgtggacgaggg-
c
cacacggtgcagtttgtgtgccgggccgatggcgacccgccgcccgccatcctctggctctcaccccgaaagca-
c
ctggtctcagccaagagcaatgggcggctcacagtcttccctgatggcacgctggaggtgcgctacgcccaggt-
a
caggacaacggcacgtacctgtgcatcgcggccaacgcgggcggcaacgactccatgcccgcccacctgcatgt-
g
cgcagctactcgcccgactggccccatcagcccaacaagaccttcgctttcatctccaaccagccgggcgaggg-
a
gaggccaacagcacccgcgccactgtgcctttccccttcgacatcaagaccctcatcatcgccaccaccatggg-
c
ttcatctctttcctgggcgtcgtcctcttctgcctggtgctgctgtttctctggagccggggcaagggcaacac-
a
aagcacaacatcgagatcgagtatgtgccccgaaagtcggacgcaggcatcagctccgccgacgcgccccgcaa-
g ttcaacatgaagatgata SEQ ID NO: 28 Cynomolgus monkey LINGO-1 mature
DNA sequence
acgggctgcccgccccgctgcgagtgctccgcccaggaccgggctgtgctctgccaccgcaagcgctttgtggc-
a
gtgcctgagggcatccccacggagacgcgcctgctggacctggggaagaaccgcatcaaaacgctcaaccagga-
c
gagttcgccagcttcccgcacctggaggagctggagctcaacgagaacatcgtgagcgccgtggagcctggcgc-
c
ttcaacaaccttttcaacctccggacgctgggtctccgcagcaaccgcctgaagctcatcccgctgggcgtctt-
c
actggcctcagcaacttgaccaagctggacatcagcgagaacaagatcgttatcctgctggactacatgttcca-
g
gacctgtacaacctcaagtcactggaggttggcgacaatgacctcgtctacatctcccaccgcgccttcagcgg-
c
ctcaacagcctggagcagctgacgctggagaaatgcaacctgacctccatccccaccgaggcgctgtcccacct-
g
cacggcctcatcgtcctgaggctccggcacctcaacatcaatgccatccgggactactccttcaagaggttgta-
c
cgactcaaggtcttggagatctcccactggccctacttggacaccatgacacccaactgcctctacggcctcaa-
c
ctgacgtccctgtccatcacgcactgcaatctgaccgctgtgccctacctggccgtccgccacctggtctatct-
c
cgcttcctcaacctctcctacaaccccatcagcaccattgagggctccatgttgcatgagctgctccggctgca-
g
gagatccagctggtgggcgggcagctggccatggtggagccctatgccttccgcggcctcaactacctgcgcgt-
g
ctcaatgtctctggcaaccagctgaccacgctggaagaatcagtcttccactcggtgggcaacctggagacgct-
c
atcctggactccaacccactggcctgcgactgtcggctcctgtgggtgttccggcgccgctggcggctcaactt-
c
aaccggcagcagcccacgtgcgccacgcccgagttcgtccagggcaaggagttcaaggacttccctgatgtgct-
a
ctgcccaactacttcacctgccgccgcgcccgcatccgggatcgcaaggcccagcaggtgtttgtggatgaggg-
c
cacacggtgcagtttgtgtgccgggccgatggcgacccgccgcccgccatcctctggctctcaccccgaaagca-
c
ctggtctcagccaagagcaatgggcggctcacagtcttccctgatggcacgctggaggtgcgctacgcccaggt-
a
caggacaatggcacgtacctgtgcatcgcggccaatgcaggcggcaacgactccatgcctgcccacctgcatgt-
g
cgcagctactcacccgactggccccatcagcccaacaagaccttcgccttcatccccaaccagccgggcgaggg-
a
gaggccaacagcacccgagccactgtgcctttccccttcgacatcaagaccctcatcatcgccaccaccatggg-
c
ttcatctctttcctgggcgtcgtcctcttctgcctggtgctgctgtttctctggagccggggcaagggcaacac-
g
aagcacaacatcgagatcgagtatgtcccccgaaagtcggacgcaggcatcagctccgccgacgcgccccgcaa-
g ttcaacatgaagatgata SEQ ID NO: 29 Rat LINGO-1 mature DNA sequence
accggctgcccgccccgctgcgagtgctcagcgcaggaccgagcagtgctctgtcaccgcaagcgctttgtggc-
g
gtgcccgagggcatccccaccgagactcgcctgctggacctgggcaaaaaccgcatcaagacactcaaccagga-
c
gagtttgccagtttcccacacctggaggagctagaactcaatgagaacattgtgagcgctgtggagccgggcgc-
c
ttcaacaacctcttcaacctgaggacgctggggcttcgcagcaaccgcctgaagctcatcccgctgggcgtctt-
c
accggcctcagcaacttgaccaagctggacatcagcgagaacaagatcgtcatcctgctagactacatgttcca-
a
gacctatacaacctcaagtcgctggaggtcggcgacaatgacctcgtctacatctcccatcgagccttcagcgg-
c
ctcaacagcctggaacagctgacgctggagaaatgcaatctgacctccatccccactgaggcactctcccacct-
g
catggcctcatcgtcctgcggctacgacacctcaacatcaatgccatacgggactactccttcaagaggctgta-
c
cgactcaaggtcttagagatctcccactggccctacctggacaccatgacccccaactgcctctacggcctcaa-
c
ctgacatccctatctatcacgcactgcaacctgacagccgtgccctatctggcagtgcgccacctggtctatct-
c
cgtttcctcaatctttcctacaaccccatcggtacaatcgagggctccatgctgcatgagctgctgcggttgca-
a
gagatccaactggtgggcgggcagctggccgtggtggagccctacgcctttcgtgggctcaattacctgcgtgt-
g
ctcaatgtttctggcaaccagctgaccaccctggaggagtcagccttccactcggtgggcaacctggagacgct-
c
attctggactccaacccactggcctgtgactgccggctgctgtgggtgttccggcgccgctggcggctcaactt-
c
aacaggcagcagcctacctgcgccacacctgagttcgtccagggcaaggagttcaaggacttccccgatgtgct-
c
ctacccaactacttcacctgccgccgggcccacatccgggaccgcaaggcacagcaggtgtttgtagatgaggg-
c
cacacggtgcagttcgtatgccgggcagatggcgaccctccaccagctatcctttggctctcaccccgcaagca-
c
ttggtctcagccaagagcaatgggcggctcacagtcttccctgatggcacgctggaggtgcgctacgcccaggt-
a
caggacaacggcacgtacctgtgcatcgcagccaatgcaggcggcaacgactccatgcccgcccacttgcatgt-
g
cgcagctactcgcctgactggccccatcaacccaacaagaccttcgccttcatctccaaccagccaggcgaggg-
a
gaggccaacagcacccgcgccactgtgcctttccccttcgacatcaagacgctcatcatcgccaccaccatggg-
c
ttcatctccttcctgggcgtggtcctattctgcctggtgctgctgtttctatggagccggggcaaaggcaacac-
a
aagcacaacatcgaaattgaatatgtgccccggaaatcggacgcaggcatcagctcagctgatgcaccccgcaa-
g ttcaacatgaagatgata SEQ ID NO: 30 Forward primer DM14
CTACGTCTAGAACGGGCTGCCCGCCCCGCT
SEQ ID NO: 31 Reverse primer DM15
GGTTTCTCGAGTCATATCATCTTCATGTTGAACTTGCGG SEQ ID NO: 32 pSecTag2-V5
vector
gacggatcgggagatctcccgatcccctatggtcgactctcagtacaatctgctctgatgccgcatagttaagc-
c
agtatctgctccctgcttgtgtgttggaggtcgctgagtagtgcgcgagcaaaatttaagctacaacaaggcaa-
g
gcttgaccgacaattgcatgaagaatctgcttagggttaggcgttttgcgctgcttcgcgatgtacgggccaga-
t
atacgcgttgacattgattattgactagttattaatagtaatcaattacggggtcattagttcatagcccatat-
a
tggagttccgcgttacataacttacggtaaatggcccgcctggctgaccgcccaacgacccccgcccattgacg-
t
caataatgacgtatgttcccatagtaacgccaatagggactttccattgacgtcaatgggtggactatttacgg-
t
aaactgcccacttggcagtacatcaagtgtatcatatgccaagtacgccccctattgacgtcaatgacggtaaa-
t
ggcccgcctggcattatgcccagtacatgaccttatgggactttcctacttggcagtacatctacgtattagtc-
a
tcgctattaccatggtgatgcggttttggcagtacatcaatgggcgtggatagcggtttgactcacggggattt-
c
caagtctccaccccattgacgtcaatgggagtttgttttggcaccaaaatcaacgggactttccaaaatgtcgt-
a
acaactccgccccattgacgcaaatgggcggtaggcgtgtacggtgggaggtctatataagcagagctctctgg-
c
taactagagaacccactgcttactggcttatcgaaattaatacgactcactatagggagacccaagctggctag-
c
caccatggagacagacacactcctgctatgggtactgctgctctgggttccaggttccactggtgacgcggccc-
a
gcccggtaagcctatccctaaccctctcctcggtctcgattctacgtctagatatcctcgagaaacccgctgat-
c
agcctcgactgtgccttctagttgccagccatctgttgtttgcccctcccccgtgccttccttgaccctggaag-
g
tgccactcccactgtcctttcctaataaaatgaggaaattgcatcgcattgtctgagtaggtgtcattctattc-
t
ggggggtggggtggggcaggacagcaagggggaggattgggaagacaatagcaggcatgctggggatgcggtgg-
g
ctctatggcttctgaggcggaaagaaccagctggggctctagggggtatccccacgcgccctgtagcggcgcat-
t
aagcgcggcgggtgtggtggttacgcgcagcgtgaccgctacacttgccagcgccctagcgcccgctcctttcg-
c
tttcttcccttcctttctcgccacgttcgccggctttccccgtcaagctctaaatcggggcatccctttagggt-
t
ccgatttagtgctttacggcacctcgaccccaaaaaacttgattagggtgatggttcacgtagtgggccatcgc-
c
ctgatagacggtttttcgccctttgacgttggagtccacgttctttaatagtggactcttgttccaaactggaa-
c
aacactcaaccctatctcggtctattcttttgatttataagggattttggggatttcggcctattggttaaaaa-
a
tgagctgatttaacaaaaatttaacgcgaattaattctgtggaatgtgtgtcagttagggtgtggaaagtcccc-
a
ggctccccagcaggcagaagtatgcaaagcatgcatctcaattagtcagcaaccaggtgtggaaagtccccagg-
c
tccccagcaggcagaagtatgcaaagcatgcatctcaattagtcagcaaccatagtcccgcccctaactccgcc-
c
atcccgcccctaactccgcccagttccgcccattctccgccccatggctgactaattttttttatttatgcaga-
g
gccgaggccgcctctgcctctgagctattccagaagtagtgaggaggcttttttggaggcctaggcttttgcaa-
a
aagctcccgggagcttgtatatccattttcggatctgatcagcacgtgttgacaattaatcatcggcatagtat-
a
tcggcatagtataatacgacaaggtgaggaactaaaccatggccaagttgaccagtgccgttccggtgctcacc-
g
cgcgcgacgtcgccggagcggtcgagttctggaccgaccggctcgggttctcccgggacttcgtggaggacgac-
t
tcgccggtgtggtccgggacgacgtgaccctgttcatcagcgcggtccaggaccaggtggtgccggacaacacc-
c
tggcctgggtgtgggtgcgcggcctggacgagctgtacgccgagtggtcggaggtcgtgtccacgaacttccgg-
g
acgcctccgggccggccatgaccgagatcggcgagcagccgtgggggcgggagttcgccctgcgcgacccggcc-
g
gcaactgcgtgcacttcgtggccgaggagcaggactgacacgtgctacgagatttcgattccaccgccgccttc-
t
atgaaaggttgggcttcggaatcgttttccgggacgccggctggatgatcctccagcgcggggatctcatgctg-
g
agttcttcgcccaccccaacttgtttattgcagcttataatggttacaaataaagcaatagcatcacaaatttc-
a
caaataaagcatttttttcactgcattctagttgtggtttgtccaaactcatcaatgtatcttatcatgtctgt-
a
taccgtcgacctctagctagagcttggcgtaatcatggtcatagctgtttcctgtgtgaaattgttatccgctc-
a
caattccacacaacatacgagccggaagcataaagtgtaaagcctggggtgcctaatgagtgagctaactcaca-
t
taattgcgttgcgctcactgcccgctttccagtcgggaaacctgtcgtgccagctgcattaatgaatcggccaa-
c
gcgcggggagaggcggtttgcgtattgggcgctcttccgcttcctcgctcactgactcgctgcgctcggtcgtt-
c
ggctgcggcgagcggtatcagctcactcaaaggcggtaatacggttatccacagaatcaggggataacgcagga-
a
agaacatgtgagcaaaaggccagcaaaaggccaggaaccgtaaaaaggccgcgttgctggcgtttttccatagg-
c
tccgcccccctgacgagcatcacaaaaatcgacgctcaagtcagaggtggcgaaacccgacaggactataaaga-
t
accaggcgtttccccctggaagctccctcgtgcgctctcctgttccgaccctgccgcttaccggatacctgtcc-
g
cctttctcccttcgggaagcgtggcgctttctcaatgctcacgctgtaggtatctcagttcggtgtaggtcgtt-
c
gctccaagctgggctgtgtgcacgaaccccccgttcagcccgaccgctgcgccttatccggtaactatcgtctt-
g
agtccaacccggtaagacacgacttatcgccactggcagcagccactggtaacaggattagcagagcgaggtat-
g
taggcggtgctacagagttcttgaagtggtggcctaactacggctacactagaaggacagtatttggtatctgc-
g
ctctgctgaagccagttaccttcggaaaaagagttggtagctcttgatccggcaaacaaaccaccgctggtagc-
g
gtggtttttttgtttgcaagcagcagattacgcgcagaaaaaaaggatctcaagaagatcctttgatcttttct-
a
cggggtctgacgctcagtggaacgaaaactcacgttaagggattttggtcatgagattatcaaaaaggatcttc-
a
cctagatccttttaaattaaaaatgaagttttaaatcaatctaaagtatatatgagtaaacttggtctgacagt-
t
accaatgcttaatcagtgaggcacctatctcagcgatctgtctatttcgttcatccatagttgcctgactcccc-
g
tcgtgtagataactacgatacgggagggcttaccatctggccccagtgctgcaatgataccgcgagacccacgc-
t
caccggctccagatttatcagcaataaaccagccagccggaagggccgagcgcagaagtggtcctgcaacttta-
t
ccgcctccatccagtctattaattgttgccgggaagctagagtaagtagttcgccagttaatagtttgcgcaac-
g
ttgttgccattgctacaggcatcgtggtgtcacgctcgtcgtttggtatggcttcattcagctccggttcccaa-
c
gatcaaggcgagttacatgatcccccatgttgtgcaaaaaagcggttagctccttcggtcctccgatcgttgtc-
a
gaagtaagttggccgcagtgttatcactcatggttatggcagcactgcataattctcttactgtcatgccatcc-
g
taagatgcttttctgtgactggtgagtactcaaccaagtcattctgagaatagtgtatgcggcgaccgagttgc-
t
cttgcccggcgtcaatacgggataataccgcgccacatagcagaactttaaaagtgctcatcattggaaaacgt-
t
cttcggggcgaaaactctcaaggatcttaccgctgttgagatccagttcgatgtaacccactcgtgcacccaac-
t
gatcttcagcatcttttactttcaccagcgtttctgggtgagcaaaaacaggaaggcaaaatgccgcaaaaaag-
g
gaataagggcgacacggaaatgttgaatactcatactcttcctttttcaatattattgaagcatttatcagggt-
t
attgtctcatgagcggatacatatttgaatgtatttagaaaaataaacaaataggggttccgcgcacatttccc-
c gaaaagtgccacctgacgtc SEQ ID NO: 33 Human LINGO-2 mature DNA
sequence
attggctgccccgctcgctgtgagtgctctgcccagaacaaatctgttagctgtcacagaaggcgattgatcgc-
c
atcccagagggcattcccatcgaaaccaaaatcttggacctcagtaaaaacaggctaaaaagcgtcaaccctga-
a
gaattcatatcatatcctctgctggaagagatagacttgagtgacaacatcattgccaatgtggaaccaggagc-
a
ttcaacaatctctttaacctgcgttccctccgcctaaaaggcaatcgtctaaagctggtccctttgggagtatt-
c
acggggctgtccaatctcactaagcttgacattagtgagaataagattgtcattttactagactacatgttcca-
a
gatctacataacctgaagtctctagaagtgggggacaatgatttggtttatatatcacacagggcattcagtgg-
g
cttcttagcttggagcagctcaccctggagaaatgcaacttaacagcagtaccaacagaagccctctcccacct-
c
cgcagcctcatcagcctgcatctgaagcatctcaatatcaacaatatgcctgtgtatgcctttaaaagattgtt-
c
cacctgaaacacctagagattgactattggcctttactggatatgatgcctgccaatagcctctacggtctcaa-
c
ctcacatccctttcagtcaccaacaccaatctgtctactgtacccttccttgcctttaaacacctggtatacct-
g
actcaccttaacctctcctacaatcccatcagcactattgaagcaggcatgttctctgacctgatccgccttca-
g
gagcttcatatagtgggggcccagcttcgcaccattgagcctcactccttccaagggctccgcttcctacgcgt-
g
ctcaatgtgtctcagaacctgctggaaactttggaagagaatgtcttctcctcccctagggctctggaggtctt-
g
agcattaacaacaaccctctggcctgtgactgccgccttctctggatcttgcagcgacagcccaccctgcagtt-
t
ggtggccagcaacctatgtgtgctggcccagacaccatccgtgagaggtctttcaaggatttccatagcactgc-
c
ctttctttttactttacctgcaaaaaacccaaaatccgtgaaaagaagttgcagcatctgctagtagatgaagg-
g
cagacagtccagctagaatgcagtgcagatggagacccgcagcctgtgatttcctgggtgacaccccgaaggcg-
t
ttcatcaccaccaagtccaatggaagagccaccgtgttgggtgatggcaccttggaaatccgctttgcccagga-
t
caagacagcgggatgtatgtttgcatcgctagcaatgctgctgggaatgataccttcacagcctccttaactgt-
g
aaaggattcgcttcagatcgttttctttatgcgaacaggacccctatgtacatgaccgactccaatgacaccat-
t
tccaatggcaccaatgccaatactttttccctggaccttaaaacaatactggtgtctacagctatgggctgctt-
c
acattcctgggagtggttttattttgttttcttctcctttttgtgtggagccgagggaaaggcaagcacaaaaa-
c
agcattgaccttgagtatgtgcccagaaaaaacaatggtgctgttgtggaaggggaggtagctggacccaggag-
g ttcaacatgaaaatgatt SEQ ID NO: 34 Forward primer DM16
CTACGTCTAGAATTGGCTGCCCCGCTCGCT SEQ ID NO: 35 Reverse primer DM17
GGTTTCTCGAGTCAAATCATTTTCATGTTGAACCTCCTG SEQ ID NO: 36 pRS5a-IgG
tcgacggatcgggagatccgggacatgtacctcccaggggcccaggaagactacgggaggctacaccaacgtca-
a
tcagaggggcctgtgtagctaccgataagcggaccctcaagagggcattagcaatagtgtttataaggccccct-
t
gttaaccctaaacgggtagcatatgcttcccgggtagtagtatatactatccagactaaccctaattcaatagc-
a
tatgttacccaacgggaagcatatgctatcgaattagggttagtaaaagggtcctaaggaacagcgatatctcc-
c
accccatgagctgtcacggttttatttacatggggtcaggattccacgagggtagtgaaccattttagtcacaa-
g
ggcagtggctgaagatcaaggagcgggcagtgaactctcctgaatcttcgcctgcttcttcattctccttcgtt-
t
agctaatagaataactgctgagttgtgaacagtaaggtgtatgtgaggtgctcgaaaacaaggtttcaggtgac-
g
cccccagaataaaatttggacggggggttcagtggtggcattgtgctatgacaccaatataaccctcacaaacc-
c
cttgggcaataaatactagtgtaggaatgaaacattctgaatatctttaacaatagaaatccatggggtgggga-
c
aagccgtaaagactggatgtccatctcacacgaatttatggctatgggcaacacataatcctagtgcaatatga-
t
actggggttattaagatgtgtcccaggcagggaccaagacaggtgaaccatgttgttacactctatttgtaaca-
a
ggggaaagagagtggacgccgacagcagcggactccactggttgtctctaacacccccgaaaattaaacggggc-
t
ccacgccaatggggcccataaacaaagacaagtggccactcttttttttgaaattgtggagtgggggcacgcgt-
c
agcccccacacgccgccctgcggttttggactgtaaaataagggtgtaataacttggctgattgtaaccccgct-
a
accactgcggtcaaaccacttgcccacaaaaccactaatggcaccccggggaatacctgcataagtaggtgggc-
g
ggccaagataggggcgcgattgctgcgatctggaggacaaattacacacacttgcgcctgagcgccaagcacag-
g
gttgttggtcctcatattcacgaggtcgctgagagcacggtgggctaatgttgccatgggtagcatatactacc-
c
aaatatctggatagcatatgctatcctaatctatatctgggtagcataggctatcctaatctatatctgggtag-
c
atatgctatcctaatctatatctgggtagtatatgctatcctaatttatatctgggtagcataggctatcctaa-
t
ctatatctgggtagcatatgctatcctaatctatatctgggtagtatatgctatcctaatctgtatccgggtag-
c
atatgctatcctaatagagattagggtagtatatgctatcctaatttatatctgggtagcatatactacccaaa-
t
atctggatagcatatgctatcctaatctatatctgggtagcatatgctatcctaatctatatctgggtagcata-
g
gctatcctaatctatatctgggtagcatatgctatcctaatctatatctgggtagtatatgctatcctaattta-
t
atctgggtagcataggctatcctaatctatatctgggtagcatatgctatcctaatctatatctgggtagtata-
t
gctatcctaatctgtatccgggtagcatatgctatcctcatgcatatacagtcagcatatgatacccagtagta-
g
agtgggagtgctatcctttgcatatgccgccacctcccaagggggcgtgaattttcgctgcttgtccttttcct-
g
catgcggatcttcaatattggccattagccatattattcattggttatatagcataaatcaatattggctattg-
g
ccattgcatacgttgtatctatatcataatatgtacatttatattggctcatgtccaatatgaccgccatgttg-
g
cattgattattgactagttattaatagtaatcaattacggggtcattagttcatagcccatatatggagttccg-
c
gttacataacttacggtaaatggcccgcctggctgaccgcccaacgacccccgcccattgacgtcaataatgac-
g
tatgttcccatagtaacgccaatagggactttccattgacgtcaatgggtggagtatttacggtaaactgccca-
c
ttggcagtacatcaagtgtatcatatgccaagtccgccccctattgacgtcaatgacggtaaatggcccgcctg-
g
cattatgcccagtacatgaccttacgggactttcctacttggcagtacatctacgtattagtcatcgctattac-
c
atggtgatgcggttttggcagtacaccaatgggcgtggatagcggtttgactcacggggatttccaagtctcca-
c
cccattgacgtcaatgggagtttgttttggcaccaaaatcaacgggactttccaaaatgtcgtaataaccccgc-
c
ccgttgacgcaaatgggcggtaggcgtgtacggtgggaggtctatataagcagagctcgtttagtgaaccgtca-
g
atcgcctggagacgccatccacgctgttttgacctccatagaagacaccgggaccgatccagcctccgcggccg-
g
gaacggtgcattggaacgcggattccccgtgccaagagtgacgtaagtaccgcctatagagtctataggcccac-
c
cccttggcttcgttagaacgcggctacaattaatacataaccttatgtatcatacacatacgatttaggtgaca-
c
tatagaataacatccactttgcctttctctccacaggtgtccactcccaggtccaactgcacggaagcttcaat-
t
gggatccctcgaggttctgttccagggtccgaaatcttgtgacaaaactcacacatgcccaccgtgcccagcac-
c
tgaactcctggggggaccgtcagtcttcctcttccccccaaaacccaaggacaccctcatgatctcccggaccc-
c
tgaggtcacatgcgtggtggtggacgtgagccacgaagaccctgaggtcaagttcaactggtacgtggacggcg-
t
ggaggtgcataatgccaagacaaagccgcgggaggagcagtacaacagcacgtaccgtgtggtcagcgtcctca-
c
cgtcctgcaccaggactggctgaatggcaaggagtacaagtgcaaggtctccaacaaagccctcccagccccca-
t
cgagaaaaccatctccaaagccaaagggcagccccgagaaccacaggtgtacaccctgcccccatcccgggatg-
a
gctgaccaagaaccaggtcagcctgacctgcctggtcaaaggcttctatcccagcgacatcgccgtggagtggg-
a
gagcaatgggcagccggagaacaactacaagaccacgcctcccgtgctggactccgacggctccttcttcctct-
a
cagcaagctcaccgtggacaagagcaggtggcagcaggggaacgtcttctcatgctccgtgatgcatgaggctc-
t
gcacaaccactacacgcagaagagcctctccctgtctccgggtaaatgagatctggtacctcgcgatggcggcc-
g
ctctagagggcccgtttaaacccgctgatcagcctcgactgtgccttctagttgccagccatctgttgtttgcc-
c
ctcccccgtgccttccttgaccctggaaggtgccactcccactgtcctttcctaataaaatgaggaaattgcat-
c
gcattgtctgagtaggtgtcattctattctggggggtggggtggggcaggacagcaagggggaggattgggaag-
a
caatagcaggcatgctggggatgcggtgggctctatggcttctgaggcggaaagaaccagctagctcgatcgag-
g
caggcagaagtatgcaaagcatgcatctcaattagtcagcaaccaggtgtggaaagtccccaggctccccagca-
g
gcagaagtatgcaaagcatgcatctcaattagtcagcaaccatagtcccgcccctaactccgcccatcccgccc-
c
taactccgcccagttccgcccattctccgccccatggctgactaattttttttatttatgcagaggccgaggcc-
g
cctctgcctctgagctattccagaagtagtgaggaggcttttttggaggcctaggcttttgcaaaaagctcccg-
g
gagcttgtatatccattttcggatctgatcagcacgtgttgacaattaatcatcggcatagtatatcggcatag-
t
ataatacgacaaggtgaggaactaaaccatggccaagttgaccagtgccgttccggtgctcaccgcgcgcgacg-
t
cgccggagcggtcgagttctggaccgaccggctcgggttctcccgggacttcgtggaggacgacttcgccggtg-
t
ggtccgggacgacgtgaccctgttcatcagcgcggtccaggaccaggtggtgccggacaacaccctggcctggg-
t
gtgggtgcgcggcctggacgagctgtacgccgagtggtcggaggtcgtgtccacgaacttccgggacgcctccg-
g
gccggccatgaccgagatcggcgagcagccgtgggggcgggagttcgccctgcgcgacccggccggcaactgcg-
t
gcacttcgtggccgaggagcaggactgacacgtgctacgagatttcgattccaccgccgccttctatgaaaggt-
t
gggcttcggaatcgttttccgggacgccggctggatgatcctccagcgcggggatctcatgctggagttcttcg-
c
ccaccccaacttgtttattgcagcttataatggttacaaataaagcaatagcatcacaaatttcacaaataaag-
c
atttttttcactgcattctagttgtggtttgtccaaactcatcaatgtatcttatcatgtctgaattttgcatt-
a
atgaatcggccaacgcgcggggagaggcggtttgcgtattgggcgctcttccgcttcctcgctcactgactcgc-
t
gcgctcggtcgttcggctgcggcgagcggtatcagctcactcaaaggcggtaatacggttatccacagaatcag-
g
ggataacgcaggaaagaacatgtgagcaaaaggccagcaaaaggccaggaaccgtaaaaaggccgcgttgctgg-
c
gtttttccataggctccgcccccctgacgagcatcacaaaaatcgacgctcaagtcagaggtggcgaaacccga-
c
aggactataaagataccaggcgtttccccctggaagctccctcgtgcgctctcctgttccgaccctgccgctta-
c
cggatacctgtccgcctttctcccttcgggaagcgtggcgctttctcaatgctcacgctgtaggtatctcagtt-
c
ggtgtaggtcgttcgctccaagctgggctgtgtgcacgaaccccccgttcagcccgaccgctgcgccttatccg-
g
taactatcgtcttgagtccaacccggtaagacacgacttatcgccactggcagcagccactggtaacaggatta-
g
cagagcgaggtatgtaggcggtgctacagagttcttgaagtggtggcctaactacggctacactagaaggacag-
t
atttggtatctgcgctctgctgaagccagttaccttcggaaaaagagttggtagctcttgatccggcaaacaaa-
c
caccgctggtagcggtggtttttttgtttgcaagcagcagattacgcgcagaaaaaaaggatctcaagaagatc-
c
tttgatcttttctacggggtctgacgctcagtggaacgaaaactcacgttaagggattttggtcatgagattat-
c
aaaaaggatcttcacctagatccttttaaattaaaaatgaagttttaaatcaatctaaagtatatatgagtaaa-
c
ttggtctgacagttaccaatgcttaatcagtgaggcacctatctcagcgatctgtctatttcgttcatccatag-
t
tgcctgactccccgtcgtgtagataactacgatacgggagggcttaccatctggccccagtgctgcaatgatac-
c
gcgagacccacgctcaccggctccagatttatcagcaataaaccagccagccggaagggccgagcgcagaagtg-
g
tcctgcaactttatccgcctccatccagtctattaattgttgccgggaagctagagtaagtagttcgccagtta-
a
tagtttgcgcaacgttgttgccattgctacaggcatcgtggtgtcacgctcgtcgtttggtatggcttcattca-
g
ctccggttcccaacgatcaaggcgagttacatgatcccccatgttgtgcaaaaaagcggttagctccttcggtc-
c
tccgatcgttgtcagaagtaagttggccgcagtgttatcactcatggttatggcagcactgcataattctctta-
c
tgtcatgccatccgtaagatgcttttctgtgactggtgagtactcaaccaagtcattctgagaatagtgtatgc-
g
gcgaccgagttgctcttgcccggcgtcaatacgggataataccgcgccacatagcagaactttaaaagtgctca-
t
cattggaaaacgttcttcggggcgaaaactctcaaggatcttaccgctgttgagatccagttcgatgtaaccca-
c
tcgtgcacccaactgatcttcagcatcttttactttcaccagcgtttctgggtgagcaaaaacaggaaggcaaa-
a
tgccgcaaaaaagggaataagggcgacacggaaatgttgaatactcatactcttcctttttcaatattattgaa-
g
catttatcagggttattgtctcatgagcggatacatatttgaatgtatttagaaaaataaacaaataggggttc-
c gcgcacatttccccgaaaagtgccacctgacgtcga SEQ ID NO: 37
natleader-hsLINGO-1-Fc/pRS5a
tcgacggatcgggagatccgggacatgtacctcccaggggcccaggaagactacgggaggctacaccaacgtca-
a
tcagaggggcctgtgtagctaccgataagcggaccctcaagagggcattagcaatagtgtttataaggccccct-
t
gttaaccctaaacgggtagcatatgcttcccgggtagtagtatatactatccagactaaccctaattcaatagc-
a
tatgttacccaacgggaagcatatgctatcgaattagggttagtaaaagggtcctaaggaacagcgatatctcc-
c
accccatgagctgtcacggttttatttacatggggtcaggattccacgagggtagtgaaccattttagtcacaa-
g
ggcagtggctgaagatcaaggagcgggcagtgaactctcctgaatcttcgcctgcttcttcattctccttcgtt-
t
agctaatagaataactgctgagttgtgaacagtaaggtgtatgtgaggtgctcgaaaacaaggtttcaggtgac-
g
cccccagaataaaatttggacggggggttcagtggtggcattgtgctatgacaccaatataaccctcacaaacc-
c
cttgggcaataaatactagtgtaggaatgaaacattctgaatatctttaacaatagaaatccatggggtgggga-
c
aagccgtaaagactggatgtccatctcacacgaatttatggctatgggcaacacataatcctagtgcaatatga-
t
actggggttattaagatgtgtcccaggcagggaccaagacaggtgaaccatgttgttacactctatttgtaaca-
a
ggggaaagagagtggacgccgacagcagcggactccactggttgtctctaacacccccgaaaattaaacggggc-
t
ccacgccaatggggcccataaacaaagacaagtggccactcttttttttgaaattgtggagtgggggcacgcgt-
c
agcccccacacgccgccctgcggttttggactgtaaaataagggtgtaataacttggctgattgtaaccccgct-
a
accactgcggtcaaaccacttgcccacaaaaccactaatggcaccccggggaatacctgcataagtaggtgggc-
g
ggccaagataggggcgcgattgctgcgatctggaggacaaattacacacacttgcgcctgagcgccaagcacag-
g
gttgttggtcctcatattcacgaggtcgctgagagcacggtgggctaatgttgccatgggtagcatatactacc-
c
aaatatctggatagcatatgctatcctaatctatatctgggtagcataggctatcctaatctatatctgggtag-
c
atatgctatcctaatctatatctgggtagtatatgctatcctaatttatatctgggtagcataggctatcctaa-
t
ctatatctgggtagcatatgctatcctaatctatatctgggtagtatatgctatcctaatctgtatccgggtag-
c
atatgctatcctaatagagattagggtagtatatgctatcctaatttatatctgggtagcatatactacccaaa-
t
atctggatagcatatgctatcctaatctatatctgggtagcatatgctatcctaatctatatctgggtagcata-
g
gctatcctaatctatatctgggtagcatatgctatcctaatctatatctgggtagtatatgctatcctaattta-
t
atctgggtagcataggctatcctaatctatatctgggtagcatatgctatcctaatctatatctgggtagtata-
t
gctatcctaatctgtatccgggtagcatatgctatcctcatgcatatacagtcagcatatgatacccagtagta-
g
agtgggagtgctatcctttgcatatgccgccacctcccaagggggcgtgaattttcgctgcttgtccttttcct-
g
catgcggatcttcaatattggccattagccatattattcattggttatatagcataaatcaatattggctattg-
g
ccattgcatacgttgtatctatatcataatatgtacatttatattggctcatgtccaatatgaccgccatgttg-
g
cattgattattgactagttattaatagtaatcaattacggggtcattagttcatagcccatatatggagttccg-
c
gttacataacttacggtaaatggcccgcctggctgaccgcccaacgacccccgcccattgacgtcaataatgac-
g
tatgttcccatagtaacgccaatagggactttccattgacgtcaatgggtggagtatttacggtaaactgccca-
c
ttggcagtacatcaagtgtatcatatgccaagtccgccccctattgacgtcaatgacggtaaatggcccgcctg-
g
cattatgcccagtacatgaccttacgggactttcctacttggcagtacatctacgtattagtcatcgctattac-
c
atggtgatgcggttttggcagtacaccaatgggcgtggatagcggtttgactcacggggatttccaagtctcca-
c
cccattgacgtcaatgggagtttgttttggcaccaaaatcaacgggactttccaaaatgtcgtaataaccccgc-
c
ccgttgacgcaaatgggcggtaggcgtgtacggtgggaggtctatataagcagagctcgtttagtgaaccgtca-
g
atcgcctggagacgccatccacgctgttttgacctccatagaagacaccgggaccgatccagcctccgcggccg-
g
gaacggtgcattggaacgcggattccccgtgccaagagtgacgtaagtaccgcctatagagtctataggcccac-
c
cccttggcttcgttagaacgcggctacaattaatacataaccttatgtatcatacacatacgatttaggtgaca-
c
tatagaataacatccactttgcctttctctccacaggtgtccactcccaggtccaactgcacggaagcttgccg-
c
caccatgcaggtgagcaagaggatgctggcggggggcgtgaggagcatgcccagccccctcctggcctgctggc-
a
gcccatcctcctgctggtgctgggctcagtgctgtcaggctcggccacgggctgcccgccccgctgcgagtgct-
c
cgcccaggaccgcgctgtgctgtgccaccgcaagcgctttgtggcagtccccgagggcatccccaccgagacgc-
g
cctgctggacctaggcaagaaccgcatcaaaacgctcaaccaggacgagttcgccagcttcccgcacctggagg-
a
gctggagctcaacgagaacatcgtgagcgccgtggagcccggcgccttcaacaacctcttcaacctccggacgc-
t
gggtctccgcagcaaccgcctgaagctcatcccgctaggcgtcttcactggcctcagcaacctgaccaagctgg-
a
catcagcgagaacaagatcgttatcctactggactacatgtttcaggacctgtacaacctcaagtcactggagg-
t
tggcgacaatgacctcgtctacatctctcaccgcgccttcagcggcctcaacagcctggagcagctgacgctgg-
a
gaaatgcaacctgacctccatccccaccgaggcgctgtcccacctgcacggcctcatcgtcctgaggctccggc-
a
cctcaacatcaatgccatccgggactactccttcaagaggctgtaccgactcaaggtcttggagatctcccact-
g
gccctacttggacaccatgacacccaactgcctctacggcctcaacctgacgtccctgtccatcacacactgca-
a
tctgaccgctgtgccctacctggccgtccgccacctagtctatctccgcttcctcaacctctcctacaacccca-
t
cagcaccattgagggctccatgttgcatgagctgctccggctgcaggagatccagctggtgggcgggcagctgg-
c
cgtggtggagccctatgccttccgcggcctcaactacctgcgcgtgctcaatgtctctggcaaccagctgacca-
c
actggaggaatcagtcttccactcggtgggcaacctggagacactcatcctggactccaacccgctggcctgcg-
a
ctgtcggctcctgtgggtgttccggcgccgctggcggctcaacttcaaccggcagcagcccacgtgcgccacgc-
c
cgagtttgtccagggcaaggagttcaaggacttccctgatgtgctactgcccaactacttcacctgccgccgcg-
c
ccgcatccgggaccgcaaggcccagcaggtgtttgtggacgagggccacacggtgcagtttgtgtgccgggccg-
a
tggcgacccgccgcccgccatcctctggctctcaccccgaaagcacctggtctcagccaagagcaatgggcggc-
t
cacagtcttccctgatggcacgctggaggtgcgctacgcccaggtacaggacaacggcacgtacctgtgcatcg-
c
ggccaacgcgggcggcaacgactccatgcccgcccacctgcatgtgcgcagctactcgcccgactggccccatc-
a
gcccaacaagaccttcgctttcatctccaaccagccgggcgagggagaggccaacagcacccgcgccactgtgc-
c
tttccccttcgacatcaagaccctcgaggttctgttccagggtccgaaatcttgtgacaaaactcacacatgcc-
c
accgtgcccagcacctgaactcctggggggaccgtcagtcttcctcttccccccaaaacccaaggacaccctca-
t
gatctcccggacccctgaggtcacatgcgtggtggtggacgtgagccacgaagaccctgaggtcaagttcaact-
g
gtacgtggacggcgtggaggtgcataatgccaagacaaagccgcgggaggagcagtacaacagcacgtaccgtg-
t
ggtcagcgtcctcaccgtcctgcaccaggactggctgaatggcaaggagtacaagtgcaaggtctccaacaaag-
c
cctcccagcccccatcgagaaaaccatctccaaagccaaagggcagccccgagaaccacaggtgtacaccctgc-
c
cccatcccgggatgagctgaccaagaaccaggtcagcctgacctgcctggtcaaaggcttctatcccagcgaca-
t
cgccgtggagtgggagagcaatgggcagccggagaacaactacaagaccacgcctcccgtgctggactccgacg-
g
ctccttcttcctctacagcaagctcaccgtggacaagagcaggtggcagcaggggaacgtcttctcatgctccg-
t
gatgcatgaggctctgcacaaccactacacgcagaagagcctctccctgtctccgggtaaatgagatctggtac-
c
tcgcgatggcggccgctctagagggcccgtttaaacccgctgatcagcctcgactgtgccttctagttgccagc-
c
atctgttgtttgcccctcccccgtgccttccttgaccctggaaggtgccactcccactgtcctttcctaataaa-
a
tgaggaaattgcatcgcattgtctgagtaggtgtcattctattctggggggtggggtggggcaggacagcaagg-
g
ggaggattgggaagacaatagcaggcatgctggggatgcggtgggctctatggcttctgaggcggaaagaacca-
g
ctagctcgatcgaggcaggcagaagtatgcaaagcatgcatctcaattagtcagcaaccaggtgtggaaagtcc-
c
caggctccccagcaggcagaagtatgcaaagcatgcatctcaattagtcagcaaccatagtcccgcccctaact-
c
cgcccatcccgcccctaactccgcccagttccgcccattctccgccccatggctgactaattttttttatttat-
g
cagaggccgaggccgcctctgcctctgagctattccagaagtagtgaggaggcttttttggaggcctaggcttt-
t
gcaaaaagctcccgggagcttgtatatccattttcggatctgatcagcacgtgttgacaattaatcatcggcat-
a
gtatatcggcatagtataatacgacaaggtgaggaactaaaccatggccaagttgaccagtgccgttccggtgc-
t
caccgcgcgcgacgtcgccggagcggtcgagttctggaccgaccggctcgggttctcccgggacttcgtggagg-
a
cgacttcgccggtgtggtccgggacgacgtgaccctgttcatcagcgcggtccaggaccaggtggtgccggaca-
a
caccctggcctgggtgtgggtgcgcggcctggacgagctgtacgccgagtggtcggaggtcgtgtccacgaact-
t
ccgggacgcctccgggccggccatgaccgagatcggcgagcagccgtgggggcgggagttcgccctgcgcgacc-
c
ggccggcaactgcgtgcacttcgtggccgaggagcaggactgacacgtgctacgagatttcgattccaccgccg-
c
cttctatgaaaggttgggcttcggaatcgttttccgggacgccggctggatgatcctccagcgcggggatctca-
t
gctggagttcttcgcccaccccaacttgtttattgcagcttataatggttacaaataaagcaatagcatcacaa-
a
tttcacaaataaagcatttttttcactgcattctagttgtggtttgtccaaactcatcaatgtatcttatcatg-
t
ctgaattttgcattaatgaatcggccaacgcgcggggagaggcggtttgcgtattgggcgctcttccgcttcct-
c
gctcactgactcgctgcgctcggtcgttcggctgcggcgagcggtatcagctcactcaaaggcggtaatacggt-
t
atccacagaatcaggggataacgcaggaaagaacatgtgagcaaaaggccagcaaaaggccaggaaccgtaaaa-
a
ggccgcgttgctggcgtttttccataggctccgcccccctgacgagcatcacaaaaatcgacgctcaagtcaga-
g
gtggcgaaacccgacaggactataaagataccaggcgtttccccctggaagctccctcgtgcgctctcctgttc-
c
gaccctgccgcttaccggatacctgtccgcctttctcccttcgggaagcgtggcgctttctcaatgctcacgct-
g
taggtatctcagttcggtgtaggtcgttcgctccaagctgggctgtgtgcacgaaccccccgttcagcccgacc-
g
ctgcgccttatccggtaactatcgtcttgagtccaacccggtaagacacgacttatcgccactggcagcagcca-
c
tggtaacaggattagcagagcgaggtatgtaggcggtgctacagagttcttgaagtggtggcctaactacggct-
a
cactagaaggacagtatttggtatctgcgctctgctgaagccagttaccttcggaaaaagagttggtagctctt-
g
atccggcaaacaaaccaccgctggtagcggtggtttttttgtttgcaagcagcagattacgcgcagaaaaaaag-
g
atctcaagaagatcctttgatcttttctacggggtctgacgctcagtggaacgaaaactcacgttaagggattt-
t
ggtcatgagattatcaaaaaggatcttcacctagatccttttaaattaaaaatgaagttttaaatcaatctaaa-
g
tatatatgagtaaacttggtctgacagttaccaatgcttaatcagtgaggcacctatctcagcgatctgtctat-
t
tcgttcatccatagttgcctgactccccgtcgtgtagataactacgatacgggagggcttaccatctggcccca-
g
tgctgcaatgataccgcgagacccacgctcaccggctccagatttatcagcaataaaccagccagccggaaggg-
c
cgagcgcagaagtggtcctgcaactttatccgcctccatccagtctattaattgttgccgggaagctagagtaa-
g
tagttcgccagttaatagtttgcgcaacgttgttgccattgctacaggcatcgtggtgtcacgctcgtcgtttg-
g
tatggcttcattcagctccggttcccaacgatcaaggcgagttacatgatcccccatgttgtgcaaaaaagcgg-
t
tagctccttcggtcctccgatcgttgtcagaagtaagttggccgcagtgttatcactcatggttatggcagcac-
t
gcataattctcttactgtcatgccatccgtaagatgcttttctgtgactggtgagtactcaaccaagtcattct-
g
agaatagtgtatgcggcgaccgagttgctcttgcccggcgtcaatacgggataataccgcgccacatagcagaa-
c
tttaaaagtgctcatcattggaaaacgttcttcggggcgaaaactctcaaggatcttaccgctgttgagatcca-
g
ttcgatgtaacccactcgtgcacccaactgatcttcagcatcttttactttcaccagcgtttctgggtgagcaa-
a
aacaggaaggcaaaatgccgcaaaaaagggaataagggcgacacggaaatgttgaatactcatactcttccttt-
t
tcaatattattgaagcatttatcagggttattgtctcatgagcggatacatatttgaatgtatttagaaaaata-
a acaaataggggttccgcgcacatttccccgaaaagtgccacctgacgtcga SEQ ID NO: 38
Igleader-hsLINGO-1-.DELTA.LRR-Fc/pRS5a
tcgacggatcgggagatccgggacatgtacctcccaggggcccaggaagactacgggaggctacaccaacgtca-
a
tcagaggggcctgtgtagctaccgataagcggaccctcaagagggcattagcaatagtgtttataaggccccct-
t
gttaaccctaaacgggtagcatatgcttcccgggtagtagtatatactatccagactaaccctaattcaatagc-
a
tatgttacccaacgggaagcatatgctatcgaattagggttagtaaaagggtcctaaggaacagcgatatctcc-
c
accccatgagctgtcacggttttatttacatggggtcaggattccacgagggtagtgaaccattttagtcacaa-
g
ggcagtggctgaagatcaaggagcgggcagtgaactctcctgaatcttcgcctgcttcttcattctccttcgtt-
t
agctaatagaataactgctgagttgtgaacagtaaggtgtatgtgaggtgctcgaaaacaaggtttcaggtgac-
g
cccccagaataaaatttggacggggggttcagtggtggcattgtgctatgacaccaatataaccctcacaaacc-
c
cttgggcaataaatactagtgtaggaatgaaacattctgaatatctttaacaatagaaatccatggggtgggga-
c
aagccgtaaagactggatgtccatctcacacgaatttatggctatgggcaacacataatcctagtgcaatatga-
t
actggggttattaagatgtgtcccaggcagggaccaagacaggtgaaccatgttgttacactctatttgtaaca-
a
ggggaaagagagtggacgccgacagcagcggactccactggttgtctctaacacccccgaaaattaaacggggc-
t
ccacgccaatggggcccataaacaaagacaagtggccactcttttttttgaaattgtggagtgggggcacgcgt-
c
agcccccacacgccgccctgcggttttggactgtaaaataagggtgtaataacttggctgattgtaaccccgct-
a
accactgcggtcaaaccacttgcccacaaaaccactaatggcaccccggggaatacctgcataagtaggtgggc-
g
ggccaagataggggcgcgattgctgcgatctggaggacaaattacacacacttgcgcctgagcgccaagcacag-
g
gttgttggtcctcatattcacgaggtcgctgagagcacggtgggctaatgttgccatgggtagcatatactacc-
c
aaatatctggatagcatatgctatcctaatctatatctgggtagcataggctatcctaatctatatctgggtag-
c
atatgctatcctaatctatatctgggtagtatatgctatcctaatttatatctgggtagcataggctatcctaa-
t
ctatatctgggtagcatatgctatcctaatctatatctgggtagtatatgctatcctaatctgtatccgggtag-
c
atatgctatcctaatagagattagggtagtatatgctatcctaatttatatctgggtagcatatactacccaaa-
t
atctggatagcatatgctatcctaatctatatctgggtagcatatgctatcctaatctatatctgggtagcata-
g
gctatcctaatctatatctgggtagcatatgctatcctaatctatatctgggtagtatatgctatcctaattta-
t
atctgggtagcataggctatcctaatctatatctgggtagcatatgctatcctaatctatatctgggtagtata-
t
gctatcctaatctgtatccgggtagcatatgctatcctcatgcatatacagtcagcatatgatacccagtagta-
g
agtgggagtgctatcctttgcatatgccgccacctcccaagggggcgtgaattttcgctgcttgtccttttcct-
g
catgcggatcttcaatattggccattagccatattattcattggttatatagcataaatcaatattggctattg-
g
ccattgcatacgttgtatctatatcataatatgtacatttatattggctcatgtccaatatgaccgccatgttg-
g
cattgattattgactagttattaatagtaatcaattacggggtcattagttcatagcccatatatggagttccg-
c
gttacataacttacggtaaatggcccgcctggctgaccgcccaacgacccccgcccattgacgtcaataatgac-
g
tatgttcccatagtaacgccaatagggactttccattgacgtcaatgggtggagtatttacggtaaactgccca-
c
ttggcagtacatcaagtgtatcatatgccaagtccgccccctattgacgtcaatgacggtaaatggcccgcctg-
g
cattatgcccagtacatgaccttacgggactttcctacttggcagtacatctacgtattagtcatcgctattac-
c
atggtgatgcggttttggcagtacaccaatgggcgtggatagcggtttgactcacggggatttccaagtctcca-
c
cccattgacgtcaatgggagtttgttttggcaccaaaatcaacgggactttccaaaatgtcgtaataaccccgc-
c
ccgttgacgcaaatgggcggtaggcgtgtacggtgggaggtctatataagcagagctcgtttagtgaaccgtca-
g
atcgcctggagacgccatccacgctgttttgacctccatagaagacaccgggaccgatccagcctccgcggccg-
g
gaacggtgcattggaacgcggattccccgtgccaagagtgacgtaagtaccgcctatagagtctataggcccac-
c
cccttggcttcgttagaacgcggctacaattaatacataaccttatgtatcatacacatacgatttaggtgaca-
c
tatagaataacatccactttgcctttctctccacaggtgtccactcccaggtccaactgcacggaagcttgccg-
c
caccatgagtgtgctcactcaggtcctggcgttgctgctgctgtggcttacaggtacgcgttgtacgggctgcc-
c
gccccgctgcgagtgctccgcccaggaccgcgctgtgctgtgccaccgcaagcgctttgtggcagtccccgagg-
g
catccccaccaacctggagacactcatcctggactccaacccgctggcctgcgactgtcggctcctgtgggtgt-
t
ccggcgccgctggcggctcaacttcaaccggcagcagcccacgtgcgccacgcccgagtttgtccagggcaagg-
a
gttcaaggacttccctgatgtgctactgcccaactacttcacctgccgccgcgcccgcatccgggaccgcaagg-
c
ccagcaggtgtttgtggacgagggccacacggtgcagtttgtgtgccgggccgatggcgacccgccgcccgcca-
t
cctctggctctcaccccgaaagcacctggtctcagccaagagcaatgggcggctcacagtcttccctgatggca-
c
gctggaggtgcgctacgcccaggtacaggacaacggcacgtacctgtgcatcgcggccaacgcgggcggcaacg-
a
ctccatgcccgcccacctgcatgtgcgcagctactcgcccgactggccccatcagcccaacaagaccttcgctt-
t
catctccaaccagccgggcgagggagaggccaacagcacccgcgccactgtgcctttccccttcgacatcaaga-
c
cctcgaggttctgttccagggtccgaaatcttgtgacaaaactcacacatgcccaccgtgcccagcacctgaac-
t
cctggggggaccgtcagtcttcctcttccccccaaaacccaaggacaccctcatgatctcccggacccctgagg-
t
cacatgcgtggtggtggacgtgagccacgaagaccctgaggtcaagttcaactggtacgtggacggcgtggagg-
t
gcataatgccaagacaaagccgcgggaggagcagtacaacagcacgtaccgtgtggtcagcgtcctcaccgtcc-
t
gcaccaggactggctgaatggcaaggagtacaagtgcaaggtctccaacaaagccctcccagcccccatcgaga-
a
aaccatctccaaagccaaagggcagccccgagaaccacaggtgtacaccctgcccccatcccgggatgagctga-
c
caagaaccaggtcagcctgacctgcctggtcaaaggcttctatcccagcgacatcgccgtggagtgggagagca-
a
tgggcagccggagaacaactacaagaccacgcctcccgtgctggactccgacggctccttcttcctctacagca-
a
gctcaccgtggacaagagcaggtggcagcaggggaacgtcttctcatgctccgtgatgcatgaggctctgcaca-
a
ccactacacgcagaagagcctctccctgtctccgggtaaatgagatctggtacctcgcgatggcggccgctcta-
g
agggcccgtttaaacccgctgatcagcctcgactgtgccttctagttgccagccatctgttgtttgcccctccc-
c
cgtgccttccttgaccctggaaggtgccactcccactgtcctttcctaataaaatgaggaaattgcatcgcatt-
g
tctgagtaggtgtcattctattctggggggtggggtggggcaggacagcaagggggaggattgggaagacaata-
g
caggcatgctggggatgcggtgggctctatggcttctgaggcggaaagaaccagctagctcgatcgaggcaggc-
a
gaagtatgcaaagcatgcatctcaattagtcagcaaccaggtgtggaaagtccccaggctccccagcaggcaga-
a
gtatgcaaagcatgcatctcaattagtcagcaaccatagtcccgcccctaactccgcccatcccgcccctaact-
c
cgcccagttccgcccattctccgccccatggctgactaattttttttatttatgcagaggccgaggccgcctct-
g
cctctgagctattccagaagtagtgaggaggcttttttggaggcctaggcttttgcaaaaagctcccgggagct-
t
gtatatccattttcggatctgatcagcacgtgttgacaattaatcatcggcatagtatatcggcatagtataat-
a
cgacaaggtgaggaactaaaccatggccaagttgaccagtgccgttccggtgctcaccgcgcgcgacgtcgccg-
g
agcggtcgagttctggaccgaccggctcgggttctcccgggacttcgtggaggacgacttcgccggtgtggtcc-
g
ggacgacgtgaccctgttcatcagcgcggtccaggaccaggtggtgccggacaacaccctggcctgggtgtggg-
t
gcgcggcctggacgagctgtacgccgagtggtcggaggtcgtgtccacgaacttccgggacgcctccgggccgg-
c
catgaccgagatcggcgagcagccgtgggggcgggagttcgccctgcgcgacccggccggcaactgcgtgcact-
t
cgtggccgaggagcaggactgacacgtgctacgagatttcgattccaccgccgccttctatgaaaggttgggct-
t
cggaatcgttttccgggacgccggctggatgatcctccagcgcggggatctcatgctggagttcttcgcccacc-
c
caacttgtttattgcagcttataatggttacaaataaagcaatagcatcacaaatttcacaaataaagcatttt-
t
ttcactgcattctagttgtggtttgtccaaactcatcaatgtatcttatcatgtctgaattttgcattaatgaa-
t
cggccaacgcgcggggagaggcggtttgcgtattgggcgctcttccgcttcctcgctcactgactcgctgcgct-
c
ggtcgttcggctgcggcgagcggtatcagctcactcaaaggcggtaatacggttatccacagaatcaggggata-
a
cgcaggaaagaacatgtgagcaaaaggccagcaaaaggccaggaaccgtaaaaaggccgcgttgctggcgtttt-
t
ccataggctccgcccccctgacgagcatcacaaaaatcgacgctcaagtcagaggtggcgaaacccgacaggac-
t
ataaagataccaggcgtttccccctggaagctccctcgtgcgctctcctgttccgaccctgccgcttaccggat-
a
cctgtccgcctttctcccttcgggaagcgtggcgctttctcaatgctcacgctgtaggtatctcagttcggtgt-
a
ggtcgttcgctccaagctgggctgtgtgcacgaaccccccgttcagcccgaccgctgcgccttatccggtaact-
a
tcgtcttgagtccaacccggtaagacacgacttatcgccactggcagcagccactggtaacaggattagcagag-
c
gaggtatgtaggcggtgctacagagttcttgaagtggtggcctaactacggctacactagaaggacagtatttg-
g
tatctgcgctctgctgaagccagttaccttcggaaaaagagttggtagctcttgatccggcaaacaaaccaccg-
c
tggtagcggtggtttttttgtttgcaagcagcagattacgcgcagaaaaaaaggatctcaagaagatcctttga-
t
cttttctacggggtctgacgctcagtggaacgaaaactcacgttaagggattttggtcatgagattatcaaaaa-
g
gatcttcacctagatccttttaaattaaaaatgaagttttaaatcaatctaaagtatatatgagtaaacttggt-
c
tgacagttaccaatgcttaatcagtgaggcacctatctcagcgatctgtctatttcgttcatccatagttgcct-
g
actccccgtcgtgtagataactacgatacgggagggcttaccatctggccccagtgctgcaatgataccgcgag-
a
cccacgctcaccggctccagatttatcagcaataaaccagccagccggaagggccgagcgcagaagtggtcctg-
c
aactttatccgcctccatccagtctattaattgttgccgggaagctagagtaagtagttcgccagttaatagtt-
t
gcgcaacgttgttgccattgctacaggcatcgtggtgtcacgctcgtcgtttggtatggcttcattcagctccg-
g
ttcccaacgatcaaggcgagttacatgatcccccatgttgtgcaaaaaagcggttagctccttcggtcctccga-
t
cgttgtcagaagtaagttggccgcagtgttatcactcatggttatggcagcactgcataattctcttactgtca-
t
gccatccgtaagatgcttttctgtgactggtgagtactcaaccaagtcattctgagaatagtgtatgcggcgac-
c
gagttgctcttgcccggcgtcaatacgggataataccgcgccacatagcagaactttaaaagtgctcatcattg-
g
aaaacgttcttcggggcgaaaactctcaaggatcttaccgctgttgagatccagttcgatgtaacccactcgtg-
c
acccaactgatcttcagcatcttttactttcaccagcgtttctgggtgagcaaaaacaggaaggcaaaatgccg-
c
aaaaaagggaataagggcgacacggaaatgttgaatactcatactcttcctttttcaatattattgaagcattt-
a
tcagggttattgtctcatgagcggatacatatttgaatgtatttagaaaaataaacaaataggggttccgcgca-
c atttccccgaaaagtgccacctgacgtcga SEQ ID NO: 39 Fab Expression
Vector pMORPH .RTM.X9_MH CTAGATAACG AGGGCAAAAA ATGAAAAAGA
CAGCTATCGC GATTGCAGTG GCACTGGCTG GTTTCGCTAC CGTAGCGCAG GCCGATATCG
TGCTGACCCA GCCGCCTTCA GTGAGTGGCG CACCAGGTCA GCGTGTGACC ATCTCGTGTA
GCGGCAGCAG CAGCAACATT GGTAATAATT ATGTGTCTTG GTACCAGCAG TTGCCCGGGA
CGGCGCCGAA ACTTCTGATT TCTCGTAATT CTAAGCGTCC CTCAGGCGTG CCGGATCGTT
TTAGCGGATC CAAAAGCGGC ACCAGCGCGA GCCTTGCGAT TACGGGCCTG CAAAGCGAAG
ACGAAGCGGA TTATTATTGC TCTACTTATG ATACTTTTTC TATTGTGTTT GGCGGCGGCA
CGAAGTTAAC CGTTCTTGGC CAGCCGAAAG CCGCACCGAG TGTGACGCTG TTTCCGCCGA
GCAGCGAAGA ATTGCAGGCG AACAAAGCGA CCCTGGTGTG CCTGATTAGC GACTTTTATC
CGGGAGCCGT GACAGTGGCC TGGAAGGCAG ATAGCAGCCC CGTCAAGGCG GGAGTGGAGA
CCACCACACC CTCCAAACAA AGCAACAACA AGTACGCGGC CAGCAGCTAT CTGAGCCTGA
CGCCTGAGCA GTGGAAGTCC CACAGAAGCT ACAGCTGCCA GGTCACGCAT GAGGGGAGCA
CCGTGGAAAA AACCGTTGCG CCGACTGAGG CCTGATAAGC ATGCGTAGGA GAAAATAAAA
TGAAACAAAG CACTATTGCA CTGGCACTCT TACCGTTGCT CTTCACCCCT GTTACCAAAG
CCCAGGTGCA ATTGCAACAG TCTGGTCCGG GCCTGGTGAA ACCGAGCCAA ACCCTGAGCC
TGACCTGTGC GATTTCCGGA GATAGCGTGA GCGATAATTC TGCTGCTTGG TCTTGGATTC
GCCAGTCTCC TGGGCGTGGC CTCGAGTGGC TGGGCCTTAT CTATCTTCGT AGCAAGTGGG
ATAACGATTA TGCGGTGAGC GTGAAAAGCC GGATTACCAT CAACCCGGAT ACTTCGAAAA
ACCAGTTTAG CCTGCAACTG AACAGCGTGA CCCCGGAAGA TACGGCCGTG TATTATTGCG
CGCGTACTGG TCGTGCTGAT GAGTTTGATG TTTGGGGCCA AGGCACCCTG GTGACGGTTA
GCTCAGCGTC GACCAAAGGT CCAAGCGTGT TTCCGCTGGC TCCGAGCAGC AAAAGCACCA
GCGGCGGCAC GGCTGCCCTG GGCTGCCTGG TTAAAGATTA TTTCCCGGAA CCAGTCACCG
TGAGCTGGAA CAGCGGGGCG CTGACCAGCG GCGTGCATAC CTTTCCGGCG GTGCTGCAAA
GCAGCGGCCT GTATAGCCTG AGCAGCGTTG TGACCGTGCC GAGCAGCAGC TTAGGCACTC
AGACCTATAT TTGCAACGTG AACCATAAAC CGAGCAACAC CAAAGTGGAT AAAAAAGTGG
AACCGAAAAG CGAATTCGAG CAGAAGCTGA TCTCTGAGGA GGATCTGAAC GGCGCGCCGC
ACCATCATCA CCATCACTGA TAAGCTTGAC CTGTGAAGTG AAAAATGGCG CAGATTGTGC
GACATTTTTT TTGTCTGCCG TTTAATTAAA GGGGGGGGGG GGCCGGCCTG GGGGGGGGTG
TACATGAAAT TGTAAACGTT AATATTTTGT TAAAATTCGC GTTAAATTTT TGTTAAATCA
GCTCATTTTT TAACCAATAG GCCGAAATCG GCAAAATCCC TTATAAATCA AAAGAATAGA
CCGAGATAGG GTTGAGTGTT GTTCCAGTTT GGAACAAGAG TCCACTATTA AAGAACGTGG
ACTCCAACGT CAAAGGGCGA AAAACCGTCT ATCAGGGCGA TGGCCCACTA CGAGAACCAT
CACCCTAATC AAGTTTTTTG GGGTCGAGGT GCCGTAAAGC ACTAAATCGG AACCCTAAAG
GGAGCCCCCG ATTTAGAGCT TGACGGGGAA AGCCGGCGAA CGTGGCGAGA AAGGAAGGGA
AGAAAGCGAA AGGAGCGGGC GCTAGGGCGC TGGCAAGTGT AGCGGTCACG CTGCGCGTAA
CCACCACACC CGCCGCGCTT AATGCGCCGC TACAGGGCGC GTGCTAGACT AGTGTTTAAA
CCGGACCGGG GGGGGGCTTA AGTGGGCTGC AAAACAAAAC GGCCTCCTGT CAGGAAGCCG
CTTTTATCGG GTAGCCTCAC TGCCCGCTTT CCAGTCGGGA AACCTGTCGT GCCAGCTGCA
TCAGTGAATC GGCCAACGCG CGGGGAGAGG CGGTTTGCGT ATTGGGAGCC AGGGTGGTTT
TTCTTTTCAC CAGTGAGACG GGCAACAGCT GATTGCCCTT CACCGCCTGG CCCTGAGAGA
GTTGCAGCAA GCGGTCCACG CTGGTTTGCC CCAGCAGGCG AAAATCCTGT TTGATGGTGG
TCAGCGGCGG GATATAACAT GAGCTGTCCT CGGTATCGTC GTATCCCACT ACCGAGATGT
CCGCACCAAC GCGCAGCCCG GACTCGGTAA TGGCACGCAT TGCGCCCAGC GCCATCTGAT
CGTTGGCAAC CAGCATCGCA GTGGGAACGA TGCCCTCATT CAGCATTTGC ATGGTTTGTT
GAAAACCGGA CATGGCACTC CAGTCGCCTT CCCGTTCCGC TATCGGCTGA ATTTGATTGC
GAGTGAGATA TTTATGCCAG CCAGCCAGAC GCAGACGCGC CGAGACAGAA CTTAATGGGC
CAGCTAACAG CGCGATTTGC TGGTGGCCCA ATGCGACCAG ATGCTCCACG CCCAGTCGCG
TACCGTCCTC ATGGGAGAAA ATAATACTGT TGATGGGTGT CTGGTCAGAG ACATCAAGAA
ATAACGCCGG AACATTAGTG CAGGCAGCTT CCACAGCAAT AGCATCCTGG TCATCCAGCG
GATAGTTAAT AATCAGCCCA CTGACACGTT GCGCGAGAAG ATTGTGCACC GCCGCTTTAC
AGGCTTCGAC GCCGCTTCGT TCTACCATCG ACACGACCAC GCTGGCACCC AGTTGATCGG
CGCGAGATTT AATCGCCGCG ACAATTTGCG ACGGCGCGTG CAGGGCCAGA CTGGAGGTGG
CAACGCCAAT CAGCAACGAC TGTTTGCCCG CCAGTTGTTG TGCCACGCGG TTAGGAATGT
AATTCAGCTC CGCCATCGCC GCTTCCACTT TTTCCCGCGT TTTCGCAGAA ACGTGGCTGG
CCTGGTTCAC CACGCGGGAA ACGGTCTGAT AAGAGACACC GGCATACTCT GCGACATCGT
ATAACGTTAC TGGTTTCACA TTCACCACCC TGAATTGACT CTCTTCCGGG CGCTATCATG
CCATACCGCG AAAGGTTTTG CGCCATTCGA TGCTAGCCAT GTGAGCAAAA GGCCAGCAAA
AGGCCAGGAA CCGTAAAAAG GCCGCGTTGC TGGCGTTTTT CCATAGGCTC CGCCCCCCTG
ACGAGCATCA CAAAAATCGA CGCTCAAGTC AGAGGTGGCG AAACCCGACA GGACTATAAA
GATACCAGGC GTTTCCCCCT GGAAGCTCCC TCGTGCGCTC TCCTGTTCCG ACCCTGCCGC
TTACCGGATA CCTGTCCGCC TTTCTCCCTT CGGGAAGCGT GGCGCTTTCT CATAGCTCAC
GCTGTAGGTA TCTCAGTTCG GTGTAGGTCG TTCGCTCCAA GCTGGGCTGT GTGCACGAAC
CCCCCGTTCA GCCCGACCGC TGCGCCTTAT CCGGTAACTA TCGTCTTGAG TCCAACCCGG
TAAGACACGA CTTATCGCCA CTGGCAGCAG CCACTGGTAA CAGGATTAGC AGAGCGAGGT
ATGTAGGCGG TGCTACAGAG TTCTTGAAGT GGTGGCCTAA CTACGGCTAC ACTAGAAGAA
CAGTATTTGG TATCTGCGCT CTGCTGTAGC CAGTTACCTT CGGAAAAAGA GTTGGTAGCT
CTTGATCCGG CAAACAAACC ACCGCTGGTA GCGGTGGTTT TTTTGTTTGC AAGCAGCAGA
TTACGCGCAG AAAAAAAGGA TCTCAAGAAG ATCCTTTGAT CTTTTCTACG GGGTCTGACG
CTCAGTGGAA CGAAAACTCA CGTTAAGGGA TTTTGGTCAG ATCTAGCACC AGGCGTTTAA
GGGCACCAAT AACTGCCTTA AAAAAATTAC GCCCCGCCCT GCCACTCATC GCAGTACTGT
TGTAATTCAT TAAGCATTCT GCCGACATGG AAGCCATCAC AAACGGCATG ATGAACCTGA
ATCGCCAGCG GCATCAGCAC CTTGTCGCCT TGCGTATAAT ATTTGCCCAT AGTGAAAACG
GGGGCGAAGA AGTTGTCCAT ATTGGCTACG TTTAAATCAA AACTGGTGAA ACTCACCCAG
GGATTGGCTG AGACGAAAAA CATATTCTCA ATAAACCCTT TAGGGAAATA GGCCAGGTTT
TCACCGTAAC ACGCCACATC TTGCGAATAT ATGTGTAGAA ACTGCCGGAA ATCGTCGTGG
TATTCACTCC AGAGCGATGA AAACGTTTCA GTTTGCTCAT GGAAAACGGT GTAACAAGGG
TGAACACTAT CCCATATCAC CAGCTCACCG TCTTTCATTG CCATACGGAA CTCCGGGTGA
GCATTCATCA GGCGGGCAAG AATGTGAATA AAGGCCGGAT AAAACTTGTG CTTATTTTTC
TTTACGGTCT TTAAAAAGGC CGTAATATCC AGCTGAACGG TCTGGTTATA GGTACATTGA
GCAACTGACT GAAATGCCTC AAAATGTTCT TTACGATGCC ATTGGGATAT ATCAACGGTG
GTATATCCAG TGATTTTTTT CTCCATTTTA GCTTCCTTAG CTCCTGAAAA TCTCGATAAC
TCAAAAAATA CGCCCGGTAG TGATCTTATT TCATTATGGT GAAAGTTGGA ACCTCACCCG
ACGTCTAATG TGAGTTAGCT CACTCATTAG GCACCCCAGG CTTTACACTT TATGCTTCCG
GCTCGTATGT TGTGTGGAAT TGTGAGCGGA TAACAATTTC ACACAGGAAA CAGCTATGAC
CATGATTACG AATTT SEQ ID NO: 40 IgG4 Expression Vector pMORPH
.RTM._h_Ig.gamma.4 AATTGCATGA AGAATCTGCT TAGGGTTAGG CGTTTTGCGC
TGCTTCGCGA TGTACGGGCC AGATATACGC GTTGACATTG ATTATTGACT AGTTATTAAT
AGTAATCAAT TACGGGGTCA TTAGTTCATA GCCCATATAT GGAGTTCCGC GTTACATAAC
TTACGGTAAA TGGCCCGCCT GGCTGACCGC CCAACGACCC CCGCCCATTG ACGTCAATAA
TGACGTATGT TCCCATAGTA ACGCCAATAG GGACTTTCCA TTGACGTCAA TGGGTGGACT
ATTTACGGTA AACTGCCCAC TTGGCAGTAC ATCAAGTGTA TCATATGCCA AGTACGCCCC
CTATTGACGT CAATGACGGT AAATGGCCCG CCTGGCATTA TGCCCAGTAC ATGACCTTAT
GGGACTTTCC TACTTGGCAG TACATCTACG TATTAGTCAT CGCTATTACC ATGGTGATGC
GGTTTTGGCA GTACATCAAT GGGCGTGGAT AGCGGTTTGA CTCACGGGGA TTTCCAAGTC
TCCACCCCAT TGACGTCAAT GGGAGTTTGT TTTGGCACCA AAATCAACGG GACTTTCCAA
AATGTCGTAA CAACTCCGCC CCATTGACGC AAATGGGCGG TAGGCGTGTA CGGTGGGAGG
TCTATATAAG CAGAGCTCTC TGGCTAACTA GAGAACCCAC TGCTTACTGG CTTATCGAAA
TTAATACGAC TCACTATAGG GAGACCCAAG CTGGCTAGCG CCACCATGAA ACACCTGTGG
TTCTTCCTCC TGCTGGTGGC AGCTCCCAGA TGGGTCCTGT CCCAGGTGGA ATTGCAACAG
TCTGGTCCGG GCCTGGTGAA ACCGAGCCAA ACCCTGAGCC TGACCTGTGC GATTTCCGGA
GATAGCGTGA GCGATAATTC TGCTGCTTGG TCTTGGATTC GCCAGTCTCC TGGGCGTGGC
CTCGAGTGGC TGGGCCTTAT CTATCTTCGT AGCAAGTGGG ATAACGATTA TGCGGTGAGC
GTGAAAAGCC GGATTACCAT CAACCCGGAT ACTTCGAAAA ACCAGTTTAG CCTGCAACTG
AACAGCGTGA CCCCGGAAGA TACGGCCGTG TATTATTGCG CGCGTACTGG TCGTGCTGAT
GAGTTTGATG TTTGGGGCCA AGGCACCCTG GTGACGGTTA GCTCAGCTTC CACCAAGGGA
CCATCCGTCT TCCCCCTGGC GCCCTGCTCC AGGAGCACCT CCGAGAGCAC AGCCGCCCTG
GGCTGCCTGG TCAAGGACTA CTTCCCCGAA CCGGTGACGG TGTCGTGGAA CTCAGGCGCC
CTGACCAGCG GCGTGCACAC CTTCCCGGCT GTCCTACAGT CCTCAGGACT CTACTCCCTC
AGCAGCGTGG TGACCGTGCC CTCCAGCAGC TTGGGCACGA AGACCTACAC CTGCAACGTA
GATCACAAGC CCAGCAACAC CAAGGTGGAC AAGAGAGTTG AGTCCAAATA TGGTCCCCCA
TGCCCATCAT GCCCAGCACC TGAGTTCCTG GGGGGACCAT CAGTCTTCCT GTTCCCCCCA
AAACCCAAGG ACACTCTCAT GATCTCCCGG ACCCCTGAGG TCACGTGCGT GGTGGTGGAC
GTGAGCCAGG AAGACCCCGA GGTCCAGTTC AACTGGTACG TGGATGGCGT GGAGGTGCAT
AATGCCAAGA CAAAGCCGCG GGAGGAGCAG TTCAACAGCA CGTACCGTGT GGTCAGCGTC
CTCACCGTCC TGCACCAGGA CTGGCTGAAC GGCAAGGAGT ACAAGTGCAA GGTCTCCAAC
AAAGGCCTCC CGTCCTCCAT CGAGAAAACC ATCTCCAAAG CCAAAGGGCA GCCCCGAGAG
CCACAGGTGT ACACCCTGCC CCCATCCCAG GAGGAGATGA CCAAGAACCA GGTCAGCCTG
ACCTGCCTGG TCAAAGGCTT CTACCCCAGC GACATCGCCG TGGAGTGGGA GAGCAATGGG
CAGCCGGAGA ACAACTACAA GACCACGCCT CCCGTGCTGG ACTCCGACGG CTCCTTCTTC
CTCTACAGCA GGCTAACCGT GGACAAGAGC AGGTGGCAGG AGGGGAATGT CTTCTCATGC
TCCGTGATGC ATGAGGCTCT GCACAACCAC TACACACAGA AGAGCCTCTC CCTGTCTCTG
GGTAAATGAG GGCCCGTTTA AACCCGCTGA TCAGCCTCGA CTGTGCCTTC TAGTTGCCAG
CCATCTGTTG TTTGCCCCTC CCCCGTGCCT TCCTTGACCC TGGAAGGTGC CACTCCCACT
GTCCTTTCCT AATAAAATGA GGAAATTGCA TCGCATTGTC TGAGTAGGTG TCATTCTATT
CTGGGGGGTG GGGTGGGGCA GGACAGCAAG GGGGAGGATT GGGAAGACAA TAGCAGGCAT
GCTGGGGATG CGGTGGGCTC TATGGCTTCT GAGGCGGAAA GAACCAGCTG GGGCTCTAGG
GGGTATCCCC ACGCGCCCTG TAGCGGCGCA TTAAGCGCGG CGGGTGTGGT GGTTACGCGC
AGCGTGACCG CTACACTTGC CAGCGCCCTA GCGCCCGCTC CTTTCGCTTT CTTCCCTTCC
TTTCTCGCCA CGTTCGCCGG CTTTCCCCGT CAAGCTCTAA ATCGGGGCAT CCCTTTAGGG
TTCCGATTTA GTGCTTTACG GCACCTCGAC CCCAAAAAAC TTGATTAGGG TGATGGTTCA
CGTAGTGGGC CATCGCCCTG ATAGACGGTT TTTCGCCCTT TGACGTTGGA GTCCACGTTC
TTTAATAGTG GACTCTTGTT CCAAACTGGA ACAACACTCA ACCCTATCTC GGTCTATTCT
TTTGATTTAT AAGGGATTTT GGGGATTTCG GCCTATTGGT TAAAAAATGA
GCTGATTTAA CAAAAATTTA ACGCGAATTA ATTCTGTGGA ATGTGTGTCA GTTAGGGTGT
GGAAAGTCCC CAGGCTCCCC AGGCAGGCAG AAGTATGCAA AGCATGCATC TCAATTAGTC
AGCAACCAGG TGTGGAAAGT CCCCAGGCTC CCCAGCAGGC AGAAGTATGC AAAGCATGCA
TCTCAATTAG TCAGCAACCA TAGTCCCGCC CCTAACTCCG CCCATCCCGC CCCTAACTCC
GCCCAGTTCC GCCCATTCTC CGCCCCATGG CTGACTAATT TTTTTTATTT ATGCAGAGGC
CGAGGCCGCC TCTGCCTCTG AGCTATTCCA GAAGTAGTGA GGAGGCTTTT TTGGAGGCCT
AGGCTTTTGC AAAAAGCTCC CGGGAGCTTG TATATCCATT TTCGGATCTG ATCAAGAGAC
AGGATGAGGA TCGTTTCGCA TGATTGAACA AGATGGATTG CACGCAGGTT CTCCGGCCGC
TTGGGTGGAG AGGCTATTCG GCTATGACTG GGCACAACAG ACAATCGGCT GCTCTGATGC
CGCCGTGTTC CGGCTGTCAG CGCAGGGGCG CCCGGTTCTT TTTGTCAAGA CCGACCTGTC
CGGTGCCCTG AATGAACTGC AGGACGAGGC AGCGCGGCTA TCGTGGCTGG CCACGACGGG
CGTTCCTTGC GCAGCTGTGC TCGACGTTGT CACTGAAGCG GGAAGGGACT GGCTGCTATT
GGGCGAAGTG CCGGGGCAGG ATCTCCTGTC ATCTCACCTT GCTCCTGCCG AGAAAGTATC
CATCATGGCT GATGCAATGC GGCGGCTGCA TACGCTTGAT CCGGCTACCT GCCCATTCGA
CCACCAAGCG AAACATCGCA TCGAGCGAGC ACGTACTCGG ATGGAAGCCG GTCTTGTCGA
TCAGGATGAT CTGGACGAAG AGCATCAGGG GCTCGCGCCA GCCGAACTGT TCGCCAGGCT
CAAGGCGCGC ATGCCCGACG GCGAGGATCT CGTCGTGACC CATGGCGATG CCTGCTTGCC
GAATATCATG GTGGAAAATG GCCGCTTTTC TGGATTCATC GACTGTGGCC GGCTGGGTGT
GGCGGACCGC TATCAGGACA TAGCGTTGGC TACCCGTGAT ATTGCTGAAG AGCTTGGCGG
CGAATGGGCT GACCGCTTCC TCGTGCTTTA CGGTATCGCC GCTCCCGATT CGCAGCGCAT
CGCCTTCTAT CGCCTTCTTG ACGAGTTCTT CTGAGCGGGA CTCTGGGGTT CGAAATGACC
GACCAAGCGA CGCCCAACCT GCCATCACGA GATTTCGATT CCACCGCCGC CTTCTATGAA
AGGTTGGGCT TCGGAATCGT TTTCCGGGAC GCCGGCTGGA TGATCCTCCA GCGCGGGGAT
CTCATGCTGG AGTTCTTCGC CCACCCCAAC TTGTTTATTG CAGCTTATAA TGGTTACAAA
TAAAGCAATA GCATCACAAA TTTCACAAAT AAAGCATTTT TTTCACTGCA TTCTAGTTGT
GGTTTGTCCA AACTCATCAA TGTATCTTAT CATGTCTGTA TACCGTCGAC CTCTAGCTAG
AGCTTGGCGT AATCATGGTC ATAGCTGTTT CCTGTGTGAA ATTGTTATCC GCTCACAATT
CCACACAACA TACGAGCCGG AAGCATAAAG TGTAAAGCCT GGGGTGCCTA ATGAGTGAGC
TAACTCACAT TAATTGCGTT GCGCTCACTG CCCGCTTTCC AGTCGGGAAA CCTGTCGTGC
CAGCTGCATT AATGAATCGG CCAACGCGCG GGGAGAGGCG GTTTGCGTAT TGGGCGCTCT
TCCGCTTCCT CGCTCACTGA CTCGCTGCGC TCGGTCGTTC GGCTGCGGCG AGCGGTATCA
GCTCACTCAA AGGCGGTAAT ACGGTTATCC ACAGAATCAG GGGATAACGC AGGAAAGAAC
ATGTGAGCAA AAGGCCAGCA AAAGGCCAGG AACCGTAAAA AGGCCGCGTT GCTGGCGTTT
TTCCATAGGC TCCGCCCCCC TGACGAGCAT CACAAAAATC GACGCTCAAG TCAGAGGTGG
CGAAACCCGA CAGGACTATA AAGATACCAG GCGTTTCCCC CTGGAAGCTC CCTCGTGCGC
TCTCCTGTTC CGACCCTGCC GCTTACCGGA TACCTGTCCG CCTTTCTCCC TTCGGGAAGC
GTGGCGCTTT CTCAATGCTC ACGCTGTAGG TATCTCAGTT CGGTGTAGGT CGTTCGCTCC
AAGCTGGGCT GTGTGCACGA ACCCCCCGTT CAGCCCGACC GCTGCGCCTT ATCCGGTAAC
TATCGTCTTG AGTCCAACCC GGTAAGACAC GACTTATCGC CACTGGCAGC AGCCACTGGT
AACAGGATTA GCAGAGCGAG GTATGTAGGC GGTGCTACAG AGTTCTTGAA GTGGTGGCCT
AACTACGGCT ACACTAGAAG GACAGTATTT GGTATCTGCG CTCTGCTGAA GCCAGTTACC
TTCGGAAAAA GAGTTGGTAG CTCTTGATCC GGCAAACAAA CCACCGCTGG TAGCGGTGGT
TTTTTTGTTT GCAAGCAGCA GATTACGCGC AGAAAAAAAG GATCTCAAGA AGATCCTTTG
ATCTTTTCTA CGGGGTCTGA CGCTCAGTGG AACGAAAACT CACGTTAAGG GATTTTGGTC
ATGAGATTAT CAAAAAGGAT CTTCACCTAG ATCCTTTTAA ATTAAAAATG AAGTTTTAAA
TCAATCTAAA GTATATATGA GTAAACTTGG TCTGACAGTT ACCAATGCTT AATCAGTGAG
GCACCTATCT CAGCGATCTG TCTATTTCGT TCATCCATAG TTGCCTGACT CCCCGTCGTG
TAGATAACTA CGATACGGGA GGGCTTACCA TCTGGCCCCA GTGCTGCAAT GATACCGCGA
GACCCACGCT CACCGGCTCC AGATTTATCA GCAATAAACC AGCCAGCCGG AAGGGCCGAG
CGCAGAAGTG GTCCTGCAAC TTTATCCGCC TCCATCCAGT CTATTAATTG TTGCCGGGAA
GCTAGAGTAA GTAGTTCGCC AGTTAATAGT TTGCGCAACG TTGTTGCCAT TGCTACAGGC
ATCGTGGTGT CACGCTCGTC GTTTGGTATG GCTTCATTCA GCTCCGGTTC CCAACGATCA
AGGCGAGTTA CATGATCCCC CATGTTGTGC AAAAAAGCGG TTAGCTCCTT CGGTCCTCCG
ATCGTTGTCA GAAGTAAGTT GGCCGCAGTG TTATCACTCA TGGTTATGGC AGCACTGCAT
AATTCTCTTA CTGTCATGCC ATCCGTAAGA TGCTTTTCTG TGACTGGTGA GTACTCAACC
AAGTCATTCT GAGAATAGTG TATGCGGCGA CCGAGTTGCT CTTGCCCGGC GTCAATACGG
GATAATACCG CGCCACATAG CAGAACTTTA AAAGTGCTCA TCATTGGAAA ACGTTCTTCG
GGGCGAAAAC TCTCAAGGAT CTTACCGCTG TTGAGATCCA GTTCGATGTA ACCCACTCGT
GCACCCAACT GATCTTCAGC ATCTTTTACT TTCACCAGCG TTTCTGGGTG AGCAAAAACA
GGAAGGCAAA ATGCCGCAAA AAAGGGAATA AGGGCGACAC GGAAATGTTG AATACTCATA
CTCTTCCTTT TTCAATATTA TTGAAGCATT TATCAGGGTT ATTGTCTCAT GAGCGGATAC
ATATTTGAAT GTATTTAGAA AAATAAACAA ATAGGGGTTC CGCGCACATT TCCCCGAAAA
GTGCCACCTG ACGTCGACGG ATCGGGAGAT CTCCCGATCC CCTATGGTCG ACTCTCAGTA
CAATCTGCTC TGATGCCGCA TAGTTAAGCC AGTATCTGCT CCCTGCTTGT GTGTTGGAGG
TCGCTGAGTA GTGCGCGAGC AAAATTTAAG CTACAACAAG GCAAGGCTTG ACCGAC SEQ
ID NO: 41 IgG Lambda Chain Expression Vector pMORPH
.RTM._h_Ig_lambda AATTGCATGA AGAATCTGCT TAGGGTTAGG CGTTTTGCGC
TGCTTCGCGA TGTACGGGCC AGATATACGC GTTGACATTG ATTATTGACT AGTTATTAAT
AGTAATCAAT TACGGGGTCA TTAGTTCATA GCCCATATAT GGAGTTCCGC GTTACATAAC
TTACGGTAAA TGGCCCGCCT GGCTGACCGC CCAACGACCC CCGCCCATTG ACGTCAATAA
TGACGTATGT TCCCATAGTA ACGCCAATAG GGACTTTCCA TTGACGTCAA TGGGTGGACT
ATTTACGGTA AACTGCCCAC TTGGCAGTAC ATCAAGTGTA TCATATGCCA AGTACGCCCC
CTATTGACGT CAATGACGGT AAATGGCCCG CCTGGCATTA TGCCCAGTAC ATGACCTTAT
GGGACTTTCC TACTTGGCAG TACATCTACG TATTAGTCAT CGCTATTACC ATGGTGATGC
GGTTTTGGCA GTACATCAAT GGGCGTGGAT AGCGGTTTGA CTCACGGGGA TTTCCAAGTC
TCCACCCCAT TGACGTCAAT GGGAGTTTGT TTTGGCACCA AAATCAACGG GACTTTCCAA
AATGTCGTAA CAACTCCGCC CCATTGACGC AAATGGGCGG TAGGCGTGTA CGGTGGGAGG
TCTATATAAG CAGAGCTCTC TGGCTAACTA GAGAACCCAC TGCTTACTGG CTTATCGAAA
TTAATACGAC TCACTATAGG GAGACCCAAG CTGGCTAGCG CCACCATGGC CTGGGCTCTG
CTGCTCCTCA CCCTCCTCAC TCAGGGCACA GGATCCTGGG CTGATATCGT GCTGACCCAG
CCGCCTTCAG TGAGTGGCGC ACCAGGTCAG CGTGTGACCA TCTCGTGTAG CGGCAGCAGC
AGCAACATTG GTAATAATTA TGTGTCTTGG TACCAGCAGT TGCCCGGGAC GGCGCCGAAA
CTTCTGATTT CTCGTAATTC TAAGCGTCCC TCAGGCGTGC CGGATCGTTT TAGCGGATCC
AAAAGCGGCA CCAGCGCGAG CCTTGCGATT ACGGGCCTGC AAAGCGAAGA CGAAGCGGAT
TATTATTGCT CTACTTATGA TACTTTTTCT ATTGTGTTTG GCGGCGGCAC GAAGTTAACC
GTCCTAGGTC AGCCCAAGGC TGCCCCCTCG GTCACTCTGT TCCCGCCCTC CTCTGAGGAG
CTTCAAGCCA ACAAGGCCAC ACTGGTGTGT CTCATAAGTG ACTTCTACCC GGGAGCCGTG
ACAGTGGCCT GGAAGGGAGA TAGCAGCCCC GTCAAGGCGG GAGTGGAGAC CACCACACCC
TCCAAACAAA GCAACAACAA GTACGCGGCC AGCAGCTATC TGAGCCTGAC GCCTGAGCAG
TGGAAGTCCC ACAGAAGCTA CAGCTGCCAG GTCACGCATG AAGGGAGCAC CGTGGAGAAG
ACAGTGGCCC CTACAGAATG TTCATAGGGG CCCGTTTAAA CCCGCTGATC AGCCTCGACT
GTGCCTTCTA GTTGCCAGCC ATCTGTTGTT TGCCCCTCCC CCGTGCCTTC CTTGACCCTG
GAAGGTGCCA CTCCCACTGT CCTTTCCTAA TAAAATGAGG AAATTGCATC GCATTGTCTG
AGTAGGTGTC ATTCTATTCT GGGGGGTGGG GTGGGGCAGG ACAGCAAGGG GGAGGATTGG
GAAGACAATA GCAGGCATGC TGGGGATGCG GTGGGCTCTA TGGCTTCTGA GGCGGAAAGA
ACCAGCTGGG GCTCTAGGGG GTATCCCCAC GCGCCCTGTA GCGGCGCATT AAGCGCGGCG
GGTGTGGTGG TTACGCGCAG CGTGACCGCT ACACTTGCCA GCGCCCTAGC GCCCGCTCCT
TTCGCTTTCT TCCCTTCCTT TCTCGCCACG TTCGCCGGCT TTCCCCGTCA AGCTCTAAAT
CGGGGCATCC CTTTAGGGTT CCGATTTAGT GCTTTACGGC ACCTCGACCC CAAAAAACTT
GATTAGGGTG ATGGTTCACG TAGTGGGCCA TCGCCCTGAT AGACGGTTTT TCGCCCTTTG
ACGTTGGAGT CCACGTTCTT TAATAGTGGA CTCTTGTTCC AAACTGGAAC AACACTCAAC
CCTATCTCGG TCTATTCTTT TGATTTATAA GGGATTTTGG GGATTTCGGC CTATTGGTTA
AAAAATGAGC TGATTTAACA AAAATTTAAC GCGAATTAAT TCTGTGGAAT GTGTGTCAGT
TAGGGTGTGG AAAGTCCCCA GGCTCCCCAG GCAGGCAGAA GTATGCAAAG CATGCATCTC
AATTAGTCAG CAACCAGGTG TGGAAAGTCC CCAGGCTCCC CAGCAGGCAG AAGTATGCAA
AGCATGCATC TCAATTAGTC AGCAACCATA GTCCCGCCCC TAACTCCGCC CATCCCGCCC
CTAACTCCGC CCAGTTCCGC CCATTCTCCG CCCCATGGCT GACTAATTTT TTTTATTTAT
GCAGAGGCCG AGGCCGCCTC TGCCTCTGAG CTATTCCAGA AGTAGTGAGG AGGCTTTTTT
GGAGGCCTAG GCTTTTGCAA AAAGCTCCCG GGAGCTTGTA TATCCATTTT CGGATCTGAT
CAGCACGTGT TGACAATTAA TCATCGGCAT AGTATATCGG CATAGTATAA TACGACAAGG
TGAGGAACTA AACCATGGCC AAGTTGACCA GTGCCGTTCC GGTGCTCACC GCGCGCGACG
TCGCCGGAGC GGTCGAGTTC TGGACCGACC GGCTCGGGTT CTCCCGGGAC TTCGTGGAGG
ACGACTTCGC CGGTGTGGTC CGGGACGACG TGACCCTGTT CATCAGCGCG GTCCAGGACC
AGGTGGTGCC GGACAACACC CTGGCCTGGG TGTGGGTGCG CGGCCTGGAC GAGCTGTACG
CCGAGTGGTC GGAGGTCGTG TCCACGAACT TCCGGGACGC CTCCGGGCCG GCCATGACCG
AGATCGGCGA GCAGCCGTGG GGGCGGGAGT TCGCCCTGCG CGACCCGGCC GGCAACTGCG
TGCACTTCGT GGCCGAGGAG CAGGACTGAC ACGTGCTACG AGATTTCGAT TCCACCGCCG
CCTTCTATGA AAGGTTGGGC TTCGGAATCG TTTTCCGGGA CGCCGGCTGG ATGATCCTCC
AGCGCGGGGA TCTCATGCTG GAGTTCTTCG CCCACCCCAA CTTGTTTATT GCAGCTTATA
ATGGTTACAA ATAAAGCAAT AGCATCACAA ATTTCACAAA TAAAGCATTT TTTTCACTGC
ATTCTAGTTG TGGTTTGTCC AAACTCATCA ATGTATCTTA TCATGTCTGT ATACCGTCGA
CCTCTAGCTA GAGCTTGGCG TAATCATGGT CATAGCTGTT TCCTGTGTGA AATTGTTATC
CGCTCACAAT TCCACACAAC ATACGAGCCG GAAGCATAAA GTGTAAAGCC TGGGGTGCCT
AATGAGTGAG CTAACTCACA TTAATTGCGT TGCGCTCACT GCCCGCTTTC CAGTCGGGAA
ACCTGTCGTG CCAGCTGCAT TAATGAATCG GCCAACGCGC GGGGAGAGGC GGTTTGCGTA
TTGGGCGCTC TTCCGCTTCC TCGCTCACTG ACTCGCTGCG CTCGGTCGTT CGGCTGCGGC
GAGCGGTATC AGCTCACTCA AAGGCGGTAA TACGGTTATC CACAGAATCA GGGGATAACG
CAGGAAAGAA CATGTGAGCA AAAGGCCAGC AAAAGGCCAG GAACCGTAAA AAGGCCGCGT
TGCTGGCGTT TTTCCATAGG CTCCGCCCCC CTGACGAGCA TCACAAAAAT CGACGCTCAA
GTCAGAGGTG GCGAAACCCG ACAGGACTAT AAAGATACCA GGCGTTTCCC CCTGGAAGCT
CCCTCGTGCG CTCTCCTGTT CCGACCCTGC CGCTTACCGG ATACCTGTCC GCCTTTCTCC
CTTCGGGAAG CGTGGCGCTT TCTCAATGCT CACGCTGTAG GTATCTCAGT TCGGTGTAGG
TCGTTCGCTC CAAGCTGGGC TGTGTGCACG AACCCCCCGT TCAGCCCGAC CGCTGCGCCT
TATCCGGTAA CTATCGTCTT GAGTCCAACC CGGTAAGACA CGACTTATCG CCACTGGCAG
CAGCCACTGG TAACAGGATT AGCAGAGCGA GGTATGTAGG CGGTGCTACA GAGTTCTTGA
AGTGGTGGCC TAACTACGGC TACACTAGAA GGACAGTATT TGGTATCTGC GCTCTGCTGA
AGCCAGTTAC CTTCGGAAAA AGAGTTGGTA GCTCTTGATC CGGCAAACAA ACCACCGCTG
GTAGCGGTGG TTTTTTTGTT TGCAAGCAGC AGATTACGCG CAGAAAAAAA GGATCTCAAG
AAGATCCTTT GATCTTTTCT ACGGGGTCTG ACGCTCAGTG GAACGAAAAC TCACGTTAAG
GGATTTTGGT CATGAGATTA TCAAAAAGGA TCTTCACCTA GATCCTTTTA AATTAAAAAT
GAAGTTTTAA ATCAATCTAA AGTATATATG AGTAAACTTG GTCTGACAGT TACCAATGCT
TAATCAGTGA GGCACCTATC TCAGCGATCT GTCTATTTCG TTCATCCATA GTTGCCTGAC
TCCCCGTCGT GTAGATAACT ACGATACGGG AGGGCTTACC ATCTGGCCCC AGTGCTGCAA
TGATACCGCG AGACCCACGC TCACCGGCTC CAGATTTATC AGCAATAAAC CAGCCAGCCG
GAAGGGCCGA GCGCAGAAGT GGTCCTGCAA CTTTATCCGC CTCCATCCAG TCTATTAATT
GTTGCCGGGA AGCTAGAGTA AGTAGTTCGC CAGTTAATAG TTTGCGCAAC GTTGTTGCCA
TTGCTACAGG CATCGTGGTG TCACGCTCGT CGTTTGGTAT GGCTTCATTC AGCTCCGGTT
CCCAACGATC AAGGCGAGTT ACATGATCCC CCATGTTGTG CAAAAAAGCG GTTAGCTCCT
TCGGTCCTCC GATCGTTGTC AGAAGTAAGT TGGCCGCAGT GTTATCACTC ATGGTTATGG
CAGCACTGCA TAATTCTCTT ACTGTCATGC CATCCGTAAG ATGCTTTTCT GTGACTGGTG
AGTACTCAAC CAAGTCATTC TGAGAATAGT GTATGCGGCG ACCGAGTTGC TCTTGCCCGG
CGTCAATACG GGATAATACC GCGCCACATA GCAGAACTTT AAAAGTGCTC ATCATTGGAA
AACGTTCTTC GGGGCGAAAA CTCTCAAGGA TCTTACCGCT GTTGAGATCC AGTTCGATGT
AACCCACTCG TGCACCCAAC TGATCTTCAG CATCTTTTAC TTTCACCAGC GTTTCTGGGT
GAGCAAAAAC AGGAAGGCAA AATGCCGCAA AAAAGGGAAT AAGGGCGACA CGGAAATGTT
GAATACTCAT ACTCTTCCTT TTTCAATATT ATTGAAGCAT TTATCAGGGT TATTGTCTCA
TGAGCGGATA CATATTTGAA TGTATTTAGA AAAATAAACA AATAGGGGTT CCGCGCACAT
TTCCCCGAAA AGTGCCACCT GACGTCGACG GATCGGGAGA TCTCCCGATC CCCTATGGTC
GACTCTCAGT ACAATCTGCT CTGATGCCGC ATAGTTAAGC CAGTATCTGC TCCCTGCTTG
TGTGTTGGAG GTCGCTGAGT AGTGCGCGAG CAAAATTTAA GCTACAACAA GGCAAGGCTT
GACCGAC SEQ ID NO: 42 IgG Kappa Chain Expression Vector pMORPH
.RTM._h_Ig_kappa AATTGCATGA AGAATCTGCT TAGGGTTAGG CGTTTTGCGC
TGCTTCGCGA TGTACGGGCC AGATATACGC GTTGACATTG ATTATTGACT AGTTATTAAT
AGTAATCAAT TACGGGGTCA TTAGTTCATA GCCCATATAT GGAGTTCCGC GTTACATAAC
TTACGGTAAA TGGCCCGCCT GGCTGACCGC CCAACGACCC CCGCCCATTG ACGTCAATAA
TGACGTATGT TCCCATAGTA ACGCCAATAG GGACTTTCCA TTGACGTCAA TGGGTGGAGT
ATTTACGGTA AACTGCCCAC TTGGCAGTAC ATCAAGTGTA TCATATGCCA AGTACGCCCC
CTATTGACGT CAATGACGGT AAATGGCCCG CCTGGCATTA TGCCCAGTAC ATGACCTTAT
GGGACTTTCC TACTTGGCAG TACATCTACG TATTAGTCAT CGCTATTACC ATGGTGATGC
GGTTTTGGCA GTACATCAAT GGGCGTGGAT AGCGGTTTGA CTCACGGGGA TTTCCAAGTC
TCCACCCCAT TGACGTCAAT GGGAGTTTGT TTTGGCACCA AAATCAACGG GACTTTCCAA
AATGTCGTAA CAACTCCGCC CCATTGACGC AAATGGGCGG TAGGCGTGTA CGGTGGGAGG
TCTATATAAG CAGAGCTCTC TGGCTAACTA GAGAACCCAC TGCTTACTGG CTTATCGAAA
TTAATACGAC TCACTATAGG GAGACCCAAG CTGGCTAGCG CCACCATGGT GTTGCAGACC
CAGGTCTTCA TTTCTCTGTT GCTCTGGATC TCTGGTGCCT ACGGGGATAT CCAGATGACC
CAGAGCCCGT CTAGCCTGAG CGCGAGCGTG GGTGATCGTG TGACCATTAC CTGCAGAGCG
AGCCAGTCTA TTTCTAATTG GCTGAATTGG TACCAGCAGA AACCAGGTAA AGCACCGAAA
CTATTAATTT ATAAGGCTTC TACTTTGCAA AGCGGGGTCC CGTCCCGTTT TAGCGGCTCT
GGATCCGGCA CTGATTTTAC CCTGACCATT AGCAGCCTGC AACCTGAAGA CTTTGCGACT
TATTATTGCC AGCAGTATGG TAATATTCCT ATTACCTTTG GCCAGGGTAC GAAAGTTGAA
ATTAAACGTA CGGTGGCTGC ACCATCTGTC TTCATCTTCC CGCCATCTGA TGAGCAGTTG
AAATCTGGAA CTGCCTCTGT TGTGTGCCTG CTGAATAACT TCTATCCCAG AGAGGCCAAA
GTACAGTGGA AGGTGGATAA CGCCCTCCAA TCGGGTAACT CCCAGGAGAG TGTCACAGAG
CAGGACAGCA AGGACAGCAC CTACAGCCTC AGCAGCACCC TGACGCTGAG CAAAGCAGAC
TACGAGAAAC ACAAAGTCTA CGCCTGCGAA GTCACCCATC AGGGCCTGAG CTCGCCCGTC
ACAAAGAGCT TCAACAGGGG AGAGTGTTAG GGGCCCGTTT AAACCCGCTG ATCAGCCTCG
ACTGTGCCTT CTAGTTGCCA GCCATCTGTT GTTTGCCCCT CCCCCGTGCC TTCCTTGACC
CTGGAAGGTG CCACTCCCAC TGTCCTTTCC TAATAAAATG AGGAAATTGC ATCGCATTGT
CTGAGTAGGT GTCATTCTAT TCTGGGGGGT GGGGTGGGGC AGGACAGCAA GGGGGAGGAT
TGGGAAGACA ATAGCAGGCA TGCTGGGGAT GCGGTGGGCT CTATGGCTTC TGAGGCGGAA
AGAACCAGCT GGGGCTCTAG GGGGTATCCC CACGCGCCCT GTAGCGGCGC ATTAAGCGCG
GCGGGTGTGG TGGTTACGCG CAGCGTGACC GCTACACTTG CCAGCGCCCT AGCGCCCGCT
CCTTTCGCTT TCTTCCCTTC CTTTCTCGCC ACGTTCGCCG GCTTTCCCCG TCAAGCTCTA
AATCGGGGGC TCCCTTTAGG GTTCCGATTT AGTGCTTTAC GGCACCTCGA CCCCAAAAAA
CTTGATTAGG GTGATGGTTC ACGTAGTGGG CCATCGCCCT GATAGACGGT TTTTCGCCCT
TTGACGTTGG AGTCCACGTT CTTTAATAGT GGACTCTTGT TCCAAACTGG AACAACACTC
AACCCTATCT CGGTCTATTC TTTTGATTTA TAAGGGATTT TGCCGATTTC GGCCTATTGG
TTAAAAAATG AGCTGATTTA ACAAAAATTT AACGCGAATT AATTCTGTGG AATGTGTGTC
AGTTAGGGTG TGGAAAGTCC CCAGGCTCCC CAGCAGGCAG AAGTATGCAA AGCATGCATC
TCAATTAGTC AGCAACCAGG TGTGGAAAGT CCCCAGGCTC CCCAGCAGGC AGAAGTATGC
AAAGCATGCA TCTCAATTAG TCAGCAACCA TAGTCCCGCC CCTAACTCCG CCCATCCCGC
CCCTAACTCC GCCCAGTTCC GCCCATTCTC CGCCCCATGG CTGACTAATT TTTTTTATTT
ATGCAGAGGC CGAGGCCGCC TCTGCCTCTG AGCTATTCCA GAAGTAGTGA GGAGGCTTTT
TTGGAGGCCT AGGCTTTTGC AAAAAGCTCC CGGGAGCTTG TATATCCATT TTCGGATCTG
ATCAGCACGT GTTGACAATT AATCATCGGC ATAGTATATC GGCATAGTAT AATACGACAA
GGTGAGGAAC TAAACCATGG CCAAGTTGAC CAGTGCCGTT CCGGTGCTCA CCGCGCGCGA
CGTCGCCGGA GCGGTCGAGT
TCTGGACCGA CCGGCTCGGG TTCTCCCGGG ACTTCGTGGA GGACGACTTC GCCGGTGTGG
TCCGGGACGA CGTGACCCTG TTCATCAGCG CGGTCCAGGA CCAGGTGGTG CCGGACAACA
CCCTGGCCTG GGTGTGGGTG CGCGGCCTGG ACGAGCTGTA CGCCGAGTGG TCGGAGGTCG
TGTCCACGAA CTTCCGGGAC GCCTCCGGGC CGGCCATGAC CGAGATCGGC GAGCAGCCGT
GGGGGCGGGA GTTCGCCCTG CGCGACCCGG CCGGCAACTG CGTGCACTTC GTGGCCGAGG
AGCAGGACTG ACACGTGCTA CGAGATTTCG ATTCCACCGC CGCCTTCTAT GAAAGGTTGG
GCTTCGGAAT CGTTTTCCGG GACGCCGGCT GGATGATCCT CCAGCGCGGG GATCTCATGC
TGGAGTTCTT CGCCCACCCC AACTTGTTTA TTGCAGCTTA TAATGGTTAC AAATAAAGCA
ATAGCATCAC AAATTTCACA AATAAAGCAT TTTTTTCACT GCATTCTAGT TGTGGTTTGT
CCAAACTCAT CAATGTATCT TATCATGTCT GTATACCGTC GACCTCTAGC TAGAGCTTGG
CGTAATCATG GTCATAGCTG TTTCCTGTGT GAAATTGTTA TCCGCTCACA ATTCCACACA
ACATACGAGC CGGAAGCATA AAGTGTAAAG CCTGGGGTGC CTAATGAGTG AGCTAACTCA
CATTAATTGC GTTGCGCTCA CTGCCCGCTT TCCAGTCGGG AAACCTGTCG TGCCAGCTGC
ATTAATGAAT CGGCCAACGC GCGGGGAGAG GCGGTTTGCG TATTGGGCGC TCTTCCGCTT
CCTCGCTCAC TGACTCGCTG CGCTCGGTCG TTCGGCTGCG GCGAGCGGTA TCAGCTCACT
CAAAGGCGGT AATACGGTTA TCCACAGAAT CAGGGGATAA CGCAGGAAAG AACATGTGAG
CAAAAGGCCA GCAAAAGGCC AGGAACCGTA AAAAGGCCGC GTTGCTGGCG TTTTTCCATA
GGCTCCGCCC CCCTGACGAG CATCACAAAA ATCGACGCTC AAGTCAGAGG TGGCGAAACC
CGACAGGACT ATAAAGATAC CAGGCGTTTC CCCCTGGAAG CTCCCTCGTG CGCTCTCCTG
TTCCGACCCT GCCGCTTACC GGATACCTGT CCGCCTTTCT CCCTTCGGGA AGCGTGGCGC
TTTCTCATAG CTCACGCTGT AGGTATCTCA GTTCGGTGTA GGTCGTTCGC TCCAAGCTGG
GCTGTGTGCA CGAACCCCCC GTTCAGCCCG ACCGCTGCGC CTTATCCGGT AACTATCGTC
TTGAGTCCAA CCCGGTAAGA CACGACTTAT CGCCACTGGC AGCAGCCACT GGTAACAGGA
TTAGCAGAGC GAGGTATGTA GGCGGTGCTA CAGAGTTCTT GAAGTGGTGG CCTAACTACG
GCTACACTAG AAGAACAGTA TTTGGTATCT GCGCTCTGCT GAAGCCAGTT ACCTTCGGAA
AAAGAGTTGG TAGCTCTTGA TCCGGCAAAC AAACCACCGC TGGTAGCGGT GGTTTTTTTG
TTTGCAAGCA GCAGATTACG CGCAGAAAAA AAGGATCTCA AGAAGATCCT TTGATCTTTT
CTACGGGGTC TGACGCTCAG TGGAACGAAA ACTCACGTTA AGGGATTTTG GTCATGAGAT
TATCAAAAAG GATCTTCACC TAGATCCTTT TAAATTAAAA ATGAAGTTTT AAATCAATCT
AAAGTATATA TGAGTAAACT TGGTCTGACA GTTACCAATG CTTAATCAGT GAGGCACCTA
TCTCAGCGAT CTGTCTATTT CGTTCATCCA TAGTTGCCTG ACTCCCCGTC GTGTAGATAA
CTACGATACG GGAGGGCTTA CCATCTGGCC CCAGTGCTGC AATGATACCG CGAGACCCAC
GCTCACCGGC TCCAGATTTA TCAGCAATAA ACCAGCCAGC CGGAAGGGCC GAGCGCAGAA
GTGGTCCTGC AACTTTATCC GCCTCCATCC AGTCTATTAA TTGTTGCCGG GAAGCTAGAG
TAAGTAGTTC GCCAGTTAAT AGTTTGCGCA ACGTTGTTGC CATTGCTACA GGCATCGTGG
TGTCACGCTC GTCGTTTGGT ATGGCTTCAT TCAGCTCCGG TTCCCAACGA TCAAGGCGAG
TTACATGATC CCCCATGTTG TGCAAAAAAG CGGTTAGCTC CTTCGGTCCT CCGATCGTTG
TCAGAAGTAA GTTGGCCGCA GTGTTATCAC TCATGGTTAT GGCAGCACTG CATAATTCTC
TTACTGTCAT GCCATCCGTA AGATGCTTTT CTGTGACTGG TGAGTACTCA ACCAAGTCAT
TCTGAGAATA GTGTATGCGG CGACCGAGTT GCTCTTGCCC GGCGTCAATA CGGGATAATA
CCGCGCCACA TAGCAGAACT TTAAAAGTGC TCATCATTGG AAAACGTTCT TCGGGGCGAA
AACTCTCAAG GATCTTACCG CTGTTGAGAT CCAGTTCGAT GTAACCCACT CGTGCACCCA
ACTGATCTTC AGCATCTTTT ACTTTCACCA GCGTTTCTGG GTGAGCAAAA ACAGGAAGGC
AAAATGCCGC AAAAAAGGGA ATAAGGGCGA CACGGAAATG TTGAATACTC ATACTCTTCC
TTTTTCAATA TTATTGAAGC ATTTATCAGG GTTATTGTCT CATGAGCGGA TACATATTTG
AATGTATTTA GAAAAATAAA CAAATAGGGG TTCCGCGCAC ATTTCCCCGA AAAGTGCCAC
CTGACGTCGA CGGATCGGGA GATCTCCCGA TCCCCTATGG TGCACTCTCA GTACAATCTG
CTCTGATGCC GCATAGTTAA GCCAGTATCT GCTCCCTGCT TGTGTGTTGG AGGTCGCTGA
GTAGTGCGCG AGCAAAATTT AAGCTACAAC AAGGCAAGGC TTGACCGAC SEQ ID NO: 43
IgG4 Expression Vector pMORPH2 .RTM._h_Ig.gamma.4 TAATACGACT
CACTATAGGG AGACCCAAGC TGGCTAGCGC CACCATGAAA CACCTGTGGT TCTTCCTCCT
GCTGGTGGCA GCTCCCAGAT GGGTCCTGTC CCAGGTGCAA TTGCAACAGT CTGGTCCGGG
CCTGGTGAAA CCGAGCCAAA CCCTGAGCCT GACCTGTGCG ATTTCCGGAG ATAGCGTGAG
CGATAATTCT GCTGCTTGGT CTTGGATTCG CCAGTCTCCT GGGCGTGGCC TCGAGTGGCT
GGGCCTTATC TATCTTCGTA GCAAGTGGGA TAACGATTAT GCGGTGAGCG TGAAAAGCCG
GATTACCATC AACCCGGATA CTTCGAAAAA CCAGTTTAGC CTGCAACTGA ACAGCGTGAC
CCCGGAAGAT ACGGCCGTGT ATTATTGCGC GCGTACTGGT CGTGCTGATG AGTTTGATGT
TTGGGGCCAA GGCACCCTGG TGACGGTTAG CTCAGCTTCC ACCAAGGGAC CATCCGTCTT
CCCCCTGGCG CCCTGCTCCA GGAGCACCTC CGAGAGCACA GCCGCCCTGG GCTGCCTGGT
CAAGGACTAC TTCCCCGAAC CGGTGACGGT GTCGTGGAAC TCAGGCGCCC TGACCAGCGG
CGTGCACACC TTCCCGGCTG TCCTACAGTC CTCAGGACTC TACTCCCTCA GCAGCGTGGT
GACCGTGCCC TCCAGCAGCT TGGGCACGAA GACCTACACC TGCAACGTAG ATCACAAGCC
CAGCAACACC AAGGTGGACA AGAGAGTTGA GTCCAAATAT GGTCCCCCAT GCCCATCATG
CCCAGCACCT GAGTTCCTGG GGGGACCATC AGTCTTCCTG TTCCCCCCAA AACCCAAGGA
CACTCTCATG ATCTCCCGGA CCCCTGAGGT CACGTGCGTG GTGGTGGACG TGAGCCAGGA
AGACCCCGAG GTCCAGTTCA ACTGGTACGT GGATGGCGTG GAGGTGCATA ATGCCAAGAC
AAAGCCGCGG GAGGAGCAGT TCAACAGCAC GTACCGTGTG GTCAGCGTCC TCACCGTCCT
GCACCAGGAC TGGCTGAACG GCAAGGAGTA CAAGTGCAAG GTCTCCAACA AAGGCCTCCC
GTCCTCCATC GAGAAAACCA TCTCCAAAGC CAAAGGGCAG CCCCGAGAGC CACAGGTGTA
CACCCTGCCC CCATCCCAGG AGGAGATGAC CAAGAACCAG GTCAGCCTGA CCTGCCTGGT
CAAAGGCTTC TACCCCAGCG ACATCGCCGT GGAGTGGGAG AGCAATGGGC AGCCGGAGAA
CAACTACAAG ACCACGCCTC CCGTGCTGGA CTCCGACGGC TCCTTCTTCC TCTACAGCAG
GCTAACCGTG GACAAGAGCA GGTGGCAGGA GGGGAATGTC TTCTCATGCT CCGTGATGCA
TGAGGCTCTG CACAACCACT ACACACAGAA GAGCCTCTCC CTGTCTCTGG GTAAATGAGG
GCCCGTTTAA ACGGGTGGCA TCCCTGTGAC CCCTCCCCAG TGCCTCTCCT GGCCCTGGAA
GTTGCCACTC CAGTGCCCAC CAGCCTTGTC CTAATAAAAT TAAGTTGCAT CATTTTGTCT
GACTAGGTGT CCTTCTATAA TATTATGGGG TGGAGGGGGG TGGTATGGAG CAAGGGGCAA
GTTGGGAAGA CAACCTGTAG GGCCTGCGGG GTCTATTGGG AACCAAGCTG GAGTGCAGTG
GCACAATCTT GGCTCACTGC AATCTCCGCC TCCTGGGTTC AAGCGATTCT CCTGCCTCAG
CCTCCCGAGT TGTTGGGATT CCAGGCATGC ATGACCAGGC TCACCTAATT TTTGTTTTTT
TGGTAGAGAC GGGGTTTCAC CATATTGGCC AGGCTGGTCT CCAACTCCTA ATCTCAGGTG
ATCTACCCAC CTTGGCCTCC CAAATTGCTG GGATTACAGG CGTGAACCAC TGCTCCCTTC
CCTGTCCTTC TGATTTTAAA ATAACTATAC CAGCAGGAGG ACGTCCAGAC ACAGCATAGG
CTACCTGGCC ATGCCCAACC GGTGGGACAT TTGAGTTGCT TGCTTGGCAC TGTCCTCTCA
TGCGTTGGGT CCACTCAGTA GATGCCTGTT GAATTGGGTA CGCGGCATCG ATTCCACGCG
CCCTGTAGCG GCGCATTAAG CGCGGCGGGT GTGGTGGTTA CGCGCAGCGT GACCGCTACA
CTTGCCAGCG CCCTAGCGCC CGCTCCTTTC GCTTTCTTCC CTTCCTTTCT CGCCACGTTC
GCCGGCTTTC CCCGTCAAGC TCTAAATCGG GGGCTCCCTT TAGGGTTCCG ATTTAGTGCT
TTACGGCACC TCGACCCCAA AAAACTTGAT TAGGGTGATG GTTCACGTAG TGGGCCATCG
CCCTGATAGA CGGTTTTTCG CCCTTTGACG TTGGAGTCCA CGTTCTTTAA TAGTGGACTC
TTGTTCCAAA CTGGAACAAC ACTCAACCCT ATCTCGGTCT ATTCTTTTGA TTTATAAGGG
ATTTTGCCGA TTTCGGCCTA TTGGTTAAAA AATGAGCTGA TTTAACAAAA ATTTAACGCG
AATTAATTCT GTGGAATGTG TGTCAGTTAG GGTGTGGAAA GTCCCCAGGC TCCCCAGCAG
GCAGAAGTAT GCAAAGCATG CATCTCAATT AGTCAGCAAC CAGGTGTGGA AAGTCCCCAG
GCTCCCCAGC AGGCAGAAGT ATGCAAAGCA TGCATCTCAA TTAGTCAGCA ACCATAGTCC
CGCCCCTAAC TCCGCCCATC CCGCCCCTAA CTCCGCCCAG TTCCGCCCAT TCTCCGCCCC
ATGGCTGACT AATTTTTTTT ATTTATGCAG AGGCCGAGGC CGCCTCTGCC TCTGAGCTAT
TCCAGAAGTA GTGAGGAGGC TTTTTTGGAG GCCTAGGCTT TTGCAAAAAG CTCCCGGGAG
CTTGTATATC CATTTTCGGA TCTGATCAAG AGACAGGATG AGGATCGTTT CGCATGATTG
AACAAGATGG ATTGCACGCA GGTTCTCCGG CCGCTTGGGT GGAGAGGCTA TTCGGCTATG
ACTGGGCACA ACAGACAATC GGCTGCTCTG ATGCCGCCGT GTTCCGGCTG TCAGCGCAGG
GGCGCCCGGT TCTTTTTGTC AAGACCGACC TGTCCGGTGC CCTGAATGAA CTGCAGGACG
AGGCAGCGCG GCTATCGTGG CTGGCCACGA CGGGCGTTCC TTGCGCAGCT GTGCTCGACG
TTGTCACTGA AGCGGGAAGG GACTGGCTGC TATTGGGCGA AGTGCCGGGG CAGGATCTCC
TGTCATCTCA CCTTGCTCCT GCCGAGAAAG TATCCATCAT GGCTGATGCA ATGCGGCGGC
TGCATACGCT TGATCCGGCT ACCTGCCCAT TCGACCACCA AGCGAAACAT CGCATCGAGC
GAGCACGTAC TCGGATGGAA GCCGGTCTTG TCGATCAGGA TGATCTGGAC GAAGAGCATC
AGGGGCTCGC GCCAGCCGAA CTGTTCGCCA GGCTCAAGGC GCGCATGCCC GACGGCGAGG
ATCTCGTCGT GACCCATGGC GATGCCTGCT TGCCGAATAT CATGGTGGAA AATGGCCGCT
TTTCTGGATT CATCGACTGT GGCCGGCTGG GTGTGGCGGA CCGCTATCAG GACATAGCGT
TGGCTACCCG TGATATTGCT GAAGAGCTTG GCGGCGAATG GGCTGACCGC TTCCTCGTGC
TTTACGGTAT CGCCGCTCCC GATTCGCAGC GCATCGCCTT CTATCGCCTT CTTGACGAGT
TCTTCTGAGC GGGACTCTGG GGTTCGAAAT GACCGACCAA GCGACGCCCA ACCTGCCATC
ACGAGATTTC GATTCCACCG CCGCCTTCTA TGAAAGGTTG GGCTTCGGAA TCGTTTTCCG
GGACGCCGGC TGGATGATCC TCCAGCGCGG GGATCTCATG CTGGAGTTCT TCGCCCACCC
CAACTTGTTT ATTGCAGCTT ATAATGGTTA CAAATAAAGC AATAGCATCA CAAATTTCAC
AAATAAAGCA TTTTTTTCAC TGCATTCTAG TTGTGGTTTG TCCAAACTCA TCAATGTATC
TTATCATGTC TGTATACCGT CGACCTCTAG CTAGAGCTTG GCGTAATCAT GGTCATAGCT
GTTTCCTGTG TGAAATTGTT ATCCGCTCAC AATTCCACAC AACATACGAG CCGGAAGCAT
AAAGTGTAAA GCCTGGGGTG CCTAATGAGT GAGCTAACTC ACATTAATTG CGTTGCGCTC
ACTGCCCGCT TTCCAGTCGG GAAACCTGTC GTGCCAGCTG CATTAATGAA TCGGCCAACG
CGCGGGGAGA GGCGGTTTGC GTATTGGGCG CTCTTCCGCT TCCTCGCTCA CTGACTCGCT
GCGCTCGGTC GTTCGGCTGC GGCGAGCGGT ATCAGCTCAC TCAAAGGCGG TAATACGGTT
ATCCACAGAA TCAGGGGATA ACGCAGGAAA GAACATGTGA GCAAAAGGCC AGCAAAAGGC
CAGGAACCGT AAAAAGGCCG CGTTGCTGGC GTTTTTCCAT AGGCTCCGCC CCCCTGACGA
GCATCACAAA AATCGACGCT CAAGTCAGAG GTGGCGAAAC CCGACAGGAC TATAAAGATA
CCAGGCGTTT CCCCCTGGAA GCTCCCTCGT GCGCTCTCCT GTTCCGACCC TGCCGCTTAC
CGGATACCTG TCCGCCTTTC TCCCTTCGGG AAGCGTGGCG CTTTCTCATA GCTCACGCTG
TAGGTATCTC AGTTCGGTGT AGGTCGTTCG CTCCAAGCTG GGCTGTGTGC ACGAACCCCC
CGTTCAGCCC GACCGCTGCG CCTTATCCGG TAACTATCGT CTTGAGTCCA ACCCGGTAAG
ACACGACTTA TCGCCACTGG CAGCAGCCAC TGGTAACAGG ATTAGCAGAG CGAGGTATGT
AGGCGGTGCT ACAGAGTTCT TGAAGTGGTG GCCTAACTAC GGCTACACTA GAAGAACAGT
ATTTGGTATC TGCGCTCTGC TGAAGCCAGT TACCTTCGGA AAAAGAGTTG GTAGCTCTTG
ATCCGGCAAA CAAACCACCG CTGGTAGCGG TGGTTTTTTT GTTTGCAAGC AGCAGATTAC
GCGCAGAAAA AAAGGATCTC AAGAAGATCC TTTGATCTTT TCTACGGGGT CTGACGCTCA
GTGGAACGAA AACTCACGTT AAGGGATTTT GGTCATGAGA TTATCAAAAA GGATCTTCAC
CTAGATCCTT TTAAATTAAA AATGAAGTTT TAAATCAATC TAAAGTATAT ATGAGTAAAC
TTGGTCTGAC AGTTACCAAT GCTTAATCAG TGAGGCACCT ATCTCAGCGA TCTGTCTATT
TCGTTCATCC ATAGTTGCCT GACTCCCCGT CGTGTAGATA ACTACGATAC GGGAGGGCTT
ACCATCTGGC CCCAGTGCTG CAATGATACC GCGAGACCCA CGCTCACCGG CTCCAGATTT
ATCAGCAATA AACCAGCCAG CCGGAAGGGC CGAGCGCAGA AGTGGTCCTG CAACTTTATC
CGCCTCCATC CAGTCTATTA ATTGTTGCCG GGAAGCTAGA GTAAGTAGTT CGCCAGTTAA
TAGTTTGCGC AACGTTGTTG CCATTGCTAC AGGCATCGTG GTGTCACGCT CGTCGTTTGG
TATGGCTTCA TTCAGCTCCG GTTCCCAACG ATCAAGGCGA GTTACATGAT CCCCCATGTT
GTGCAAAAAA GCGGTTAGCT CCTTCGGTCC TCCGATCGTT GTCAGAAGTA AGTTGGCCGC
AGTGTTATCA CTCATGGTTA TGGCAGCACT GCATAATTCT CTTACTGTCA TGCCATCCGT
AAGATGCTTT TCTGTGACTG GTGAGTACTC AACCAAGTCA TTCTGAGAAT AGTGTATGCG
GCGACCGAGT TGCTCTTGCC CGGCGTCAAT ACGGGATAAT ACCGCGCCAC ATAGCAGAAC
TTTAAAAGTG CTCATCATTG GAAAACGTTC TTCGGGGCGA AAACTCTCAA GGATCTTACC
GCTGTTGAGA TCCAGTTCGA TGTAACCCAC TCGTGCACCC AACTGATCTT CAGCATCTTT
TACTTTCACC AGCGTTTCTG GGTGAGCAAA AACAGGAAGG CAAAATGCCG CAAAAAAGGG
AATAAGGGCG ACACGGAAAT GTTGAATACT CATACTCTTC CTTTTTCAAT ATTATTGAAG
CATTTATCAG GGTTATTGTC TCATGAGCGG ATACATATTT GAATGTATTT AGAAAAATAA
ACAAATAGGG GTTCCGCGCA CATTTCCCCG AAAAGTGCCA CCTGACGTCG ACGGATCGGG
AGATCTCCCG ATCCCCTATG GTGCACTCTC AGTACAATCT GCTCTGATGC CGCATAGTTA
AGCCAGTATC TGCTCCCTGC TTGTGTGTTG GAGGTCGCTG AGTAGTGCGC GAGCAAAATT
TAAGCTACAA CAAGGCAAGG CTTGACCGAC ATTTGCATGA AGAATCTGCT TAGGGTTAGG
CGTTTTGCGC TGCTTCGCGA TGTACGGGCC AGATATACGC GTTGACATTG ATTATTGACT
AGTTATTAAT AGTAATCAAT TACGGGGTCA TTAGTTCATA GCCCATATAT GGAGTTCCGC
GTTACATAAC TTACGGTAAA TGGCCCGCCT GGCTGACCGC CCAACGACCC CCGCCCATTG
ACGTCAATAA TGACGTATGT TCCCATAGTA ACGCCAATAG GGACTTTCCA TTGACGTCAA
TGGGTGGAGT ATTTACGGTA AACTGCCCAC TTGGCAGTAC ATCAAGTGTA TCATATGCCA
AGTACGCCCC CTATTGACGT CAATGACGGT AAATGGCCCG CCTGGCATTA TGCCCAGTAC
ATGACCTTAT GGGACTTTCC TACTTGGCAG TACATCTACG TATTAGTCAT CGCTATTACC
ATGGTGATGC GGTTTTGGCA GTACATCAAT GGGCGTGGAT AGCGGTTTGA CTCACGGGGA
TTTCCAAGTC TCCACCCCAT TGACGTCAAT GGGAGTTTGT TTTGGCACCA AAATCAACGG
GACTTTCCAA AATGTCGTAA CAACTCCGCC CCATTGACGC AAATGGGCGG TAGGCGTGTA
CGGTGGGAGG TCTATATAAG CAGAGCTCTC TGGCTAACTA GAGAACCCAC TGCTTACTGG
CTTATCGAAA T SEQ ID NO: 44 IgG Lambda Chain Expression Vector
pMORPH .RTM.2_h_Ig_lambda2 TAATACGACT CACTATAGGG AGACCCAAGC
TGGCTAGCGC CACCATGGCC TGGGCTCTGC TGCTCCTCAC CCTCCTCACT CAGGGCACAG
GATCCTGGGC TGATATCGTG CTGACCCAGC CGCCTTCAGT GAGTGGCGCA CCAGGTCAGC
GTGTGACCAT CTCGTGTAGC GGCAGCAGCA GCAACATTGG TAATAATTAT GTGTCTTGGT
ACCAGCAGTT GCCCGGGACG GCGCCGAAAC TTCTGATTTC TCGTAATTCT AAGCGTCCCT
CAGGCGTGCC GGATCGTTTT AGCGGATCCA AAAGCGGCAC CAGCGCGAGC CTTGCGATTA
CGGGCCTGCA AAGCGAAGAC GAAGCGGATT ATTATTGCTC TACTTATGAT ACTTTTTCTA
TTGTGTTTGG CGGCGGCACG AAGTTAACCG TCCTAGGTCA GCCCAAGGCT GCCCCCTCGG
TCACTCTGTT CCCGCCCTCC TCTGAGGAGC TTCAAGCCAA CAAGGCCACA CTGGTGTGTC
TCATAAGTGA CTTCTACCCG GGAGCCGTGA CAGTGGCCTG GAAGGCAGAT AGCAGCCCCG
TCAAGGCGGG AGTGGAGACC ACCACACCCT CCAAACAAAG CAACAACAAG TACGCGGCCA
GCAGCTATCT GAGCCTGACG CCTGAGCAGT GGAAGTCCCA CAGAAGCTAC AGCTGCCAGG
TCACGCATGA AGGGAGCACC GTGGAGAAGA CAGTGGCCCC TACAGAATGT TCATAGGGGC
CCGTTTAAAC GGGTGGCATC CCTGTGACCC CTCCCCAGTG CCTCTCCTGG CCCTGGAAGT
TGCCACTCCA GTGCCCACCA GCCTTGTCCT AATAAAATTA AGTTGCATCA TTTTGTCTGA
CTAGGTGTCC TTCTATAATA TTATGGGGTG GAGGGGGGTG GTATGGAGCA AGGGGCAAGT
TGGGAAGACA ACCTGTAGGG CCTGCGGGGT CTATTGGGAA CCAAGCTGGA GTGCAGTGGC
ACAATCTTGG CTCACTGCAA TCTCCGCCTC CTGGGTTCAA GCGATTCTCC TGCCTCAGCC
TCCCGAGTTG TTGGGATTCC AGGCATGCAT GACCAGGCTC ACCTAATTTT TGTTTTTTTG
GTAGAGACGG GGTTTCACCA TATTGGCCAG GCTGGTCTCC AACTCCTAAT CTCAGGTGAT
CTACCCACCT TGGCCTCCCA AATTGCTGGG ATTACAGGCG TGAACCACTG CTCCCTTCCC
TGTCCTTCTG ATTTTAAAAT AACTATACCA GCAGGAGGAC GTCCAGACAC AGCATAGGCT
ACCTGGCCAT GCCCAACCGG TGGGACATTT GAGTTGCTTG CTTGGCACTG TCCTCTCATG
CGTTGGGTCC ACTCAGTAGA TGCCTGTTGA ATTGGGTACG CGGCATCGAT TCCACGCGCC
CTGTAGCGGC GCATTAAGCG CGGCGGGTGT GGTGGTTACG CGCAGCGTGA CCGCTACACT
TGCCAGCGCC CTAGCGCCCG CTCCTTTCGC TTTCTTCCCT TCCTTTCTCG CCACGTTCGC
CGGCTTTCCC CGTCAAGCTC TAAATCGGGG GCTCCCTTTA GGGTTCCGAT TTAGTGCTTT
ACGGCACCTC GACCCCAAAA AACTTGATTA GGGTGATGGT TCACGTAGTG GGCCATCGCC
CTGATAGACG GTTTTTCGCC CTTTGACGTT GGAGTCCACG TTCTTTAATA GTGGACTCTT
GTTCCAAACT GGAACAACAC TCAACCCTAT CTCGGTCTAT TCTTTTGATT TATAAGGGAT
TTTGGGGATT TCGGCCTATT GGTTAAAAAA TGAGCTGATT TAACAAAAAT TTAACGCGAA
TTAATTCTGT GGAATGTGTG TCAGTTAGGG TGTGGAAAGT CCCCAGGCTC CCCAGGCAGG
CAGAAGTATG CAAAGCATGC ATCTCAATTA GTCAGCAACC AGGTGTGGAA AGTCCCCAGG
CTCCCCAGCA GGCAGAAGTA TGCAAAGCAT GCATCTCAAT TAGTCAGCAA CCATAGTCCC
GCCCCTAACT CCGCCCATCC CGCCCCTAAC TCCGCCCAGT TCCGCCCATT CTCCGCCCCA
TGGCTGACTA ATTTTTTTTA TTTATGCAGA GGCCGAGGCC GCCTCTGCCT CTGAGCTATT
CCAGAAGTAG TGAGGAGGCT
TTTTTGGAGG CCTAGGCTTT TGCAAAAAGC TCCCGGGAGC TTGTATATCC ATTTTCGGAT
CTGATCAGCA CGTGTTGACA ATTAATCATC GGCATAGTAT ATCGGCATAG TATAATACGA
CAAGGTGAGG AACTAAACCA TGGCCAAGTT GACCAGTGCC GTTCCGGTGC TCACCGCGCG
CGACGTCGCC GGAGCGGTCG AGTTCTGGAC CGACCGGCTC GGGTTCTCCC GGGACTTCGT
GGAGGACGAC TTCGCCGGTG TGGTCCGGGA CGACGTGACC CTGTTCATCA GCGCGGTCCA
GGACCAGGTG GTGCCGGACA ACACCCTGGC CTGGGTGTGG GTGCGCGGCC TGGACGAGCT
GTACGCCGAG TGGTCGGAGG TCGTGTCCAC GAACTTCCGG GACGCCTCCG GGCCGGCCAT
GACCGAGATC GGCGAGCAGC CGTGGGGGCG GGAGTTCGCC CTGCGCGACC CGGCCGGCAA
CTGCGTGCAC TTCGTGGCCG AGGAGCAGGA CTGACACGTG CTACGAGATT TCGATTCCAC
CGCCGCCTTC TATGAAAGGT TGGGCTTCGG AATCGTTTTC CGGGACGCCG GCTGGATGAT
CCTCCAGCGC GGGGATCTCA TGCTGGAGTT CTTCGCCCAC CCCAACTTGT TTATTGCAGC
TTATAATGGT TACAAATAAA GCAATAGCAT CACAAATTTC ACAAATAAAG CATTTTTTTC
ACTGCATTCT AGTTGTGGTT TGTCCAAACT CATCAATGTA TCTTATCATG TCTGTATACC
GTCGACCTCT AGCTAGAGCT TGGCGTAATC ATGGTCATAG CTGTTTCCTG TGTGAAATTG
TTATCCGCTC ACAATTCCAC ACAACATACG AGCCGGAAGC ATAAAGTGTA AAGCCTGGGG
TGCCTAATGA GTGAGCTAAC TCACATTAAT TGCGTTGCGC TCACTGCCCG CTTTCCAGTC
GGGAAACCTG TCGTGCCAGC TGCATTAATG AATCGGCCAA CGCGCGGGGA GAGGCGGTTT
GCGTATTGGG CGCTCTTCCG CTTCCTCGCT CACTGACTCG CTGCGCTCGG TCGTTCGGCT
GCGGCGAGCG GTATCAGCTC ACTCAAAGGC GGTAATACGG TTATCCACAG AATCAGGGGA
TAACGCAGGA AAGAACATGT GAGCAAAAGG CCAGCAAAAG GCCAGGAACC GTAAAAAGGC
CGCGTTGCTG GCGTTTTTCC ATAGGCTCCG CCCCCCTGAC GAGCATCACA AAAATCGACG
CTCAAGTCAG AGGTGGCGAA ACCCGACAGG ACTATAAAGA TACCAGGCGT TTCCCCCTGG
AAGCTCCCTC GTGCGCTCTC CTGTTCCGAC CCTGCCGCTT ACCGGATACC TGTCCGCCTT
TCTCCCTTCG GGAAGCGTGG CGCTTTCTCA ATGCTCACGC TGTAGGTATC TCAGTTCGGT
GTAGGTCGTT CGCTCCAAGC TGGGCTGTGT GCACGAACCC CCCGTTCAGC CCGACCGCTG
CGCCTTATCC GGTAACTATC GTCTTGAGTC CAACCCGGTA AGACACGACT TATCGCCACT
GGCAGCAGCC ACTGGTAACA GGATTAGCAG AGCGAGGTAT GTAGGCGGTG CTACAGAGTT
CTTGAAGTGG TGGCCTAACT ACGGCTACAC TAGAAGGACA GTATTTGGTA TCTGCGCTCT
GCTGAAGCCA GTTACCTTCG GAAAAAGAGT TGGTAGCTCT TGATCCGGCA AACAAACCAC
CGCTGGTAGC GGTGGTTTTT TTGTTTGCAA GCAGCAGATT ACGCGCAGAA AAAAAGGATC
TCAAGAAGAT CCTTTGATCT TTTCTACGGG GTCTGACGCT CAGTGGAACG AAAACTCACG
TTAAGGGATT TTGGTCATGA GATTATCAAA AAGGATCTTC ACCTAGATCC TTTTAAATTA
AAAATGAAGT TTTAAATCAA TCTAAAGTAT ATATGAGTAA ACTTGGTCTG ACAGTTACCA
ATGCTTAATC AGTGAGGCAC CTATCTCAGC GATCTGTCTA TTTCGTTCAT CCATAGTTGC
CTGACTCCCC GTCGTGTAGA TAACTACGAT ACGGGAGGGC TTACCATCTG GCCCCAGTGC
TGCAATGATA CCGCGAGACC CACGCTCACC GGCTCCAGAT TTATCAGCAA TAAACCAGCC
AGCCGGAAGG GCCGAGCGCA GAAGTGGTCC TGCAACTTTA TCCGCCTCCA TCCAGTCTAT
TAATTGTTGC CGGGAAGCTA GAGTAAGTAG TTCGCCAGTT AATAGTTTGC GCAACGTTGT
TGCCATTGCT ACAGGCATCG TGGTGTCACG CTCGTCGTTT GGTATGGCTT CATTCAGCTC
CGGTTCCCAA CGATCAAGGC GAGTTACATG ATCCCCCATG TTGTGCAAAA AAGCGGTTAG
CTCCTTCGGT CCTCCGATCG TTGTCAGAAG TAAGTTGGCC GCAGTGTTAT CACTCATGGT
TATGGCAGCA CTGCATAATT CTCTTACTGT CATGCCATCC GTAAGATGCT TTTCTGTGAC
TGGTGAGTAC TCAACCAAGT CATTCTGAGA ATAGTGTATG CGGCGACCGA GTTGCTCTTG
CCCGGCGTCA ATACGGGATA ATACCGCGCC ACATAGCAGA ACTTTAAAAG TGCTCATCAT
TGGAAAACGT TCTTCGGGGC GAAAACTCTC AAGGATCTTA CCGCTGTTGA GATCCAGTTC
GATGTAACCC ACTCGTGCAC CCAACTGATC TTCAGCATCT TTTACTTTCA CCAGCGTTTC
TGGGTGAGCA AAAACAGGAA GGCAAAATGC CGCAAAAAAG GGAATAAGGG CGACACGGAA
ATGTTGAATA CTCATACTCT TCCTTTTTCA ATATTATTGA AGCATTTATC AGGGTTATTG
TCTCATGAGC GGATACATAT TTGAATGTAT TTAGAAAAAT AAACAAATAG GGGTTCCGCG
CACATTTCCC CGAAAAGTGC CACCTGACGT CGACGGATCG GGAGATCTCC CGATCCCCTA
TGGTCGACTC TCAGTACAAT CTGCTCTGAT GCCGCATAGT TAAGCCAGTA TCTGCTCCCT
GCTTGTGTGT TGGAGGTCGC TGAGTAGTGC GCGAGCAAAA TTTAAGCTAC AACAAGGCAA
GGCTTGACCG ACAATTGCAT GAAGAATCTG CTTAGGGTTA GGCGTTTTGC GCTGCTTCGC
GATGTACGGG CCAGATATAC GCGTTGACAT TGATTATTGA CTAGTTATTA ATAGTAATCA
ATTACGGGGT CATTAGTTCA TAGCCCATAT ATGGAGTTCC GCGTTACATA ACTTACGGTA
AATGGCCCGC CTGGCTGACC GCCCAACGAC CCCCGCCCAT TGACGTCAAT AATGACGTAT
GTTCCCATAG TAACGCCAAT AGGGACTTTC CATTGACGTC AATGGGTGGA CTATTTACGG
TAAACTGCCC ACTTGGCAGT ACATCAAGTG TATCATATGC CAAGTACGCC CCCTATTGAC
GTCAATGACG GTAAATGGCC CGCCTGGCAT TATGCCCAGT ACATGACCTT ATGGGACTTT
CCTACTTGGC AGTACATCTA CGTATTAGTC ATCGCTATTA CCATGGTGAT GCGGTTTTGG
CAGTACATCA ATGGGCGTGG ATAGCGGTTT GACTCACGGG GATTTCCAAG TCTCCACCCC
ATTGACGTCA ATGGGAGTTT GTTTTGGCAC CAAAATCAAC GGGACTTTCC AAAATGTCGT
AACAACTCCG CCCCATTGAC GCAAATGGGC GGTAGGCGTG TACGGTGGGA GGTCTATATA
AGCAGAGCTC TCTGGCTAAC TAGAGAACCC ACTGCTTACT GGCTTATCGA AAT SEQ ID
NO: 45 IgG kappa Chain Expression Vector pMORPH .RTM.2_h_Ig_kappa
TAATACGACT CACTATAGGG AGACCCAAGC TGGCTAGCGC CACCATGGTG TTGCAGACCC
AGGTCTTCAT TTCTCTGTTG CTCTGGATCT CTGGTGCCTA CGGGGATATC CAGATGACCC
AGAGCCCGTC TAGCCTGAGC GCGAGCGTGG GTGATCGTGT GACCATTACC TGCAGAGCGA
GCCAGTCTAT TTCTAATTGG CTGAATTGGT ACCAGCAGAA ACCAGGTAAA GCACCGAAAC
TATTAATTTA TAAGGCTTCT ACTTTGCAAA GCGGGGTCCC GTCCCGTTTT AGCGGCTCTG
GATCCGGCAC TGATTTTACC CTGACCATTA GCAGCCTGCA ACCTGAAGAC TTTGCGACTT
ATTATTGCCA GCAGTATGGT AATATTCCTA TTACCTTTGG CCAGGGTACG AAAGTTGAAA
TTAAACGTAC GGTGGCTGCA CCATCTGTCT TCATCTTCCC GCCATCTGAT GAGCAGTTGA
AATCTGGAAC TGCCTCTGTT GTGTGCCTGC TGAATAACTT CTATCCCAGA GAGGCCAAAG
TACAGTGGAA GGTGGATAAC GCCCTCCAAT CGGGTAACTC CCAGGAGAGT GTCACAGAGC
AGGACAGCAA GGACAGCACC TACAGCCTCA GCAGCACCCT GACGCTGAGC AAAGCAGACT
ACGAGAAACA CAAAGTCTAC GCCTGCGAAG TCACCCATCA GGGCCTGAGC TCGCCCGTCA
CAAAGAGCTT CAACAGGGGA GAGTGTTAGG GGCCCGTTTA AACGGGTGGC ATCCCTGTGA
CCCCTCCCCA GTGCCTCTCC TGGCCCTGGA AGTTGCCACT CCAGTGCCCA CCAGCCTTGT
CCTAATAAAA TTAAGTTGCA TCATTTTGTC TGACTAGGTG TCCTTCTATA ATATTATGGG
GTGGAGGGGG GTGGTATGGA GCAAGGGGCA AGTTGGGAAG ACAACCTGTA GGGCCTGCGG
GGTCTATTGG GAACCAAGCT GGAGTGCAGT GGCACAATCT TGGCTCACTG CAATCTCCGC
CTCCTGGGTT CAAGCGATTC TCCTGCCTCA GCCTCCCGAG TTGTTGGGAT TCCAGGCATG
CATGACCAGG CTCACCTAAT TTTTGTTTTT TTGGTAGAGA CGGGGTTTCA CCATATTGGC
CAGGCTGGTC TCCAACTCCT AATCTCAGGT GATCTACCCA CCTTGGCCTC CCAAATTGCT
GGGATTACAG GCGTGAACCA CTGCTCCCTT CCCTGTCCTT CTGATTTTAA AATAACTATA
CCAGCAGGAG GACGTCCAGA CACAGCATAG GCTACCTGGC CATGCCCAAC CGGTGGGACA
TTTGAGTTGC TTGCTTGGCA CTGTCCTCTC ATGCGTTGGG TCCACTCAGT AGATGCCTGT
TGAATTGGGT ACGCGGCATC GATTCCACGC GCCCTGTAGC GGCGCATTAA GCGCGGCGGG
TGTGGTGGTT ACGCGCAGCG TGACCGCTAC ACTTGCCAGC GCCCTAGCGC CCGCTCCTTT
CGCTTTCTTC CCTTCCTTTC TCGCCACGTT CGCCGGCTTT CCCCGTCAAG CTCTAAATCG
GGGGCTCCCT TTAGGGTTCC GATTTAGTGC TTTACGGCAC CTCGACCCCA AAAAACTTGA
TTAGGGTGAT GGTTCACGTA GTGGGCCATC GCCCTGATAG ACGGTTTTTC GCCCTTTGAC
GTTGGAGTCC ACGTTCTTTA ATAGTGGACT CTTGTTCCAA ACTGGAACAA CACTCAACCC
TATCTCGGTC TATTCTTTTG ATTTATAAGG GATTTTGGGG ATTTCGGCCT ATTGGTTAAA
AAATGAGCTG ATTTAACAAA AATTTAACGC GAATTAATTC TGTGGAATGT GTGTCAGTTA
GGGTGTGGAA AGTCCCCAGG CTCCCCAGGC AGGCAGAAGT ATGCAAAGCA TGCATCTCAA
TTAGTCAGCA ACCAGGTGTG GAAAGTCCCC AGGCTCCCCA GCAGGCAGAA GTATGCAAAG
CATGCATCTC AATTAGTCAG CAACCATAGT CCCGCCCCTA ACTCCGCCCA TCCCGCCCCT
AACTCCGCCC AGTTCCGCCC ATTCTCCGCC CCATGGCTGA CTAATTTTTT TTATTTATGC
AGAGGCCGAG GCCGCCTCTG CCTCTGAGCT ATTCCAGAAG TAGTGAGGAG GCTTTTTTGG
AGGCCTAGGC TTTTGCAAAA AGCTCCCGGG AGCTTGTATA TCCATTTTCG GATCTGATCA
GCACGTGTTG ACAATTAATC ATCGGCATAG TATATCGGCA TAGTATAATA CGACAAGGTG
AGGAACTAAA CCATGGCCAA GTTGACCAGT GCCGTTCCGG TGCTCACCGC GCGCGACGTC
GCCGGAGCGG TCGAGTTCTG GACCGACCGG CTCGGGTTCT CCCGGGACTT CGTGGAGGAC
GACTTCGCCG GTGTGGTCCG GGACGACGTG ACCCTGTTCA TCAGCGCGGT CCAGGACCAG
GTGGTGCCGG ACAACACCCT GGCCTGGGTG TGGGTGCGCG GCCTGGACGA GCTGTACGCC
GAGTGGTCGG AGGTCGTGTC CACGAACTTC CGGGACGCCT CCGGGCCGGC CATGACCGAG
ATCGGCGAGC AGCCGTGGGG GCGGGAGTTC GCCCTGCGCG ACCCGGCCGG CAACTGCGTG
CACTTCGTGG CCGAGGAGCA GGACTGACAC GTGCTACGAG ATTTCGATTC CACCGCCGCC
TTCTATGAAA GGTTGGGCTT CGGAATCGTT TTCCGGGACG CCGGCTGGAT GATCCTCCAG
CGCGGGGATC TCATGCTGGA GTTCTTCGCC CACCCCAACT TGTTTATTGC AGCTTATAAT
GGTTACAAAT AAAGCAATAG CATCACAAAT TTCACAAATA AAGCATTTTT TTCACTGCAT
TCTAGTTGTG GTTTGTCCAA ACTCATCAAT GTATCTTATC ATGTCTGTAT ACCGTCGACC
TCTAGCTAGA GCTTGGCGTA ATCATGGTCA TAGCTGTTTC CTGTGTGAAA TTGTTATCCG
CTCACAATTC CACACAACAT ACGAGCCGGA AGCATAAAGT GTAAAGCCTG GGGTGCCTAA
TGAGTGAGCT AACTCACATT AATTGCGTTG CGCTCACTGC CCGCTTTCCA GTCGGGAAAC
CTGTCGTGCC AGCTGCATTA ATGAATCGGC CAACGCGCGG GGAGAGGCGG TTTGCGTATT
GGGCGCTCTT CCGCTTCCTC GCTCACTGAC TCGCTGCGCT CGGTCGTTCG GCTGCGGCGA
GCGGTATCAG CTCACTCAAA GGCGGTAATA CGGTTATCCA CAGAATCAGG GGATAACGCA
GGAAAGAACA TGTGAGCAAA AGGCCAGCAA AAGGCCAGGA ACCGTAAAAA GGCCGCGTTG
CTGGCGTTTT TCCATAGGCT CCGCCCCCCT GACGAGCATC ACAAAAATCG ACGCTCAAGT
CAGAGGTGGC GAAACCCGAC AGGACTATAA AGATACCAGG CGTTTCCCCC TGGAAGCTCC
CTCGTGCGCT CTCCTGTTCC GACCCTGCCG CTTACCGGAT ACCTGTCCGC CTTTCTCCCT
TCGGGAAGCG TGGCGCTTTC TCAATGCTCA CGCTGTAGGT ATCTCAGTTC GGTGTAGGTC
GTTCGCTCCA AGCTGGGCTG TGTGCACGAA CCCCCCGTTC AGCCCGACCG CTGCGCCTTA
TCCGGTAACT ATCGTCTTGA GTCCAACCCG GTAAGACACG ACTTATCGCC ACTGGCAGCA
GCCACTGGTA ACAGGATTAG CAGAGCGAGG TATGTAGGCG GTGCTACAGA GTTCTTGAAG
TGGTGGCCTA ACTACGGCTA CACTAGAAGG ACAGTATTTG GTATCTGCGC TCTGCTGAAG
CCAGTTACCT TCGGAAAAAG AGTTGGTAGC TCTTGATCCG GCAAACAAAC CACCGCTGGT
AGCGGTGGTT TTTTTGTTTG CAAGCAGCAG ATTACGCGCA GAAAAAAAGG ATCTCAAGAA
GATCCTTTGA TCTTTTCTAC GGGGTCTGAC GCTCAGTGGA ACGAAAACTC ACGTTAAGGG
ATTTTGGTCA TGAGATTATC AAAAAGGATC TTCACCTAGA TCCTTTTAAA TTAAAAATGA
AGTTTTAAAT CAATCTAAAG TATATATGAG TAAACTTGGT CTGACAGTTA CCAATGCTTA
ATCAGTGAGG CACCTATCTC AGCGATCTGT CTATTTCGTT CATCCATAGT TGCCTGACTC
CCCGTCGTGT AGATAACTAC GATACGGGAG GGCTTACCAT CTGGCCCCAG TGCTGCAATG
ATACCGCGAG ACCCACGCTC ACCGGCTCCA GATTTATCAG CAATAAACCA GCCAGCCGGA
AGGGCCGAGC GCAGAAGTGG TCCTGCAACT TTATCCGCCT CCATCCAGTC TATTAATTGT
TGCCGGGAAG CTAGAGTAAG TAGTTCGCCA GTTAATAGTT TGCGCAACGT TGTTGCCATT
GCTACAGGCA TCGTGGTGTC ACGCTCGTCG TTTGGTATGG CTTCATTCAG CTCCGGTTCC
CAACGATCAA GGCGAGTTAC ATGATCCCCC ATGTTGTGCA AAAAAGCGGT TAGCTCCTTC
GGTCCTCCGA TCGTTGTCAG AAGTAAGTTG GCCGCAGTGT TATCACTCAT GGTTATGGCA
GCACTGCATA ATTCTCTTAC TGTCATGCCA TCCGTAAGAT GCTTTTCTGT GACTGGTGAG
TACTCAACCA AGTCATTCTG AGAATAGTGT ATGCGGCGAC CGAGTTGCTC TTGCCCGGCG
TCAATACGGG ATAATACCGC GCCACATAGC AGAACTTTAA AAGTGCTCAT CATTGGAAAA
CGTTCTTCGG GGCGAAAACT CTCAAGGATC TTACCGCTGT TGAGATCCAG TTCGATGTAA
CCCACTCGTG CACCCAACTG ATCTTCAGCA TCTTTTACTT TCACCAGCGT TTCTGGGTGA
GCAAAAACAG GAAGGCAAAA TGCCGCAAAA AAGGGAATAA GGGCGACACG GAAATGTTGA
ATACTCATAC TCTTCCTTTT TCAATATTAT TGAAGCATTT ATCAGGGTTA TTGTCTCATG
AGCGGATACA TATTTGAATG TATTTAGAAA AATAAACAAA TAGGGGTTCC GCGCACATTT
CCCCGAAAAG TGCCACCTGA CGTCGACGGA TCGGGAGATC TCCCGATCCC CTATGGTCGA
CTCTCAGTAC AATCTGCTCT GATGCCGCAT AGTTAAGCCA GTATCTGCTC CCTGCTTGTG
TGTTGGAGGT CGCTGAGTAG TGCGCGAGCA AAATTTAAGC TACAACAAGG CAAGGCTTGA
CCGACAATTG CATGAAGAAT CTGCTTAGGG TTAGGCGTTT TGCGCTGCTT CGCGATGTAC
GGGCCAGATA TACGCGTTGA CATTGATTAT TGACTAGTTA TTAATAGTAA TCAATTACGG
GGTCATTAGT TCATAGCCCA TATATGGAGT TCCGCGTTAC ATAACTTACG GTAAATGGCC
CGCCTGGCTG ACCGCCCAAC GACCCCCGCC CATTGACGTC AATAATGACG TATGTTCCCA
TAGTAACGCC AATAGGGACT TTCCATTGAC GTCAATGGGT GGACTATTTA CGGTAAACTG
CCCACTTGGC AGTACATCAA GTGTATCATA TGCCAAGTAC GCCCCCTATT GACGTCAATG
ACGGTAAATG GCCCGCCTGG CATTATGCCC AGTACATGAC CTTATGGGAC TTTCCTACTT
GGCAGTACAT CTACGTATTA GTCATCGCTA TTACCATGGT GATGCGGTTT TGGCAGTACA
TCAATGGGCG TGGATAGCGG TTTGACTCAC GGGGATTTCC AAGTCTCCAC CCCATTGACG
TCAATGGGAG TTTGTTTTGG CACCAAAATC AACGGGACTT TCCAAAATGT CGTAACAACT
CCGCCCCATT GACGCAAATG GGCGGTAGGC GTGTACGGTG GGAGGTCTAT ATAAGCAGAG
CTCTCTGGCT AACTAGAGAA CCCACTGCTT ACTGGCTTAT CGAAAT
Sequence CWU 1
1
511517PRTRattus sp. 1Thr Gly Cys Pro Pro Arg Cys Glu Cys Ser Ala
Gln Asp Arg Ala Val 1 5 10 15 Leu Cys His Arg Lys Arg Phe Val Ala
Val Pro Glu Gly Ile Pro Thr 20 25 30 Glu Thr Arg Leu Leu Asp Leu
Gly Lys Asn Arg Ile Lys Thr Leu Asn 35 40 45 Gln Asp Glu Phe Ala
Ser Phe Pro His Leu Glu Glu Leu Glu Leu Asn 50 55 60 Glu Asn Ile
Val Ser Ala Val Glu Pro Gly Ala Phe Asn Asn Leu Phe 65 70 75 80 Asn
Leu Arg Thr Leu Gly Leu Arg Ser Asn Arg Leu Lys Leu Ile Pro 85 90
95 Leu Gly Val Phe Thr Gly Leu Ser Asn Leu Thr Lys Leu Asp Ile Ser
100 105 110 Glu Asn Lys Ile Val Ile Leu Leu Asp Tyr Met Phe Gln Asp
Leu Tyr 115 120 125 Asn Leu Lys Ser Leu Glu Val Gly Asp Asn Asp Leu
Val Tyr Ile Ser 130 135 140 His Arg Ala Phe Ser Gly Leu Asn Ser Leu
Glu Gln Leu Thr Leu Glu 145 150 155 160 Lys Cys Asn Leu Thr Ser Ile
Pro Thr Glu Ala Leu Ser His Leu His 165 170 175 Gly Leu Ile Val Leu
Arg Leu Arg His Leu Asn Ile Asn Ala Ile Arg 180 185 190 Asp Tyr Ser
Phe Lys Arg Leu Tyr Arg Leu Lys Val Leu Glu Ile Ser 195 200 205 His
Trp Pro Tyr Leu Asp Thr Met Thr Pro Asn Cys Leu Tyr Gly Leu 210 215
220 Asn Leu Thr Ser Leu Ser Ile Thr His Cys Asn Leu Thr Ala Val Pro
225 230 235 240 Tyr Leu Ala Val Arg His Leu Val Tyr Leu Arg Phe Leu
Asn Leu Ser 245 250 255 Tyr Asn Pro Ile Gly Thr Ile Glu Gly Ser Met
Leu His Glu Leu Leu 260 265 270 Arg Leu Gln Glu Ile Gln Leu Val Gly
Gly Gln Leu Ala Val Val Glu 275 280 285 Pro Tyr Ala Phe Arg Gly Leu
Asn Tyr Leu Arg Val Leu Asn Val Ser 290 295 300 Gly Asn Gln Leu Thr
Thr Leu Glu Glu Ser Ala Phe His Ser Val Gly 305 310 315 320 Asn Leu
Glu Thr Leu Ile Leu Asp Ser Asn Pro Leu Ala Cys Asp Cys 325 330 335
Arg Leu Leu Trp Val Phe Arg Arg Arg Trp Arg Leu Asn Phe Asn Arg 340
345 350 Gln Gln Pro Thr Cys Ala Thr Pro Glu Phe Val Gln Gly Lys Glu
Phe 355 360 365 Lys Asp Phe Pro Asp Val Leu Leu Pro Asn Tyr Phe Thr
Cys Arg Arg 370 375 380 Ala His Ile Arg Asp Arg Lys Ala Gln Gln Val
Phe Val Asp Glu Gly 385 390 395 400 His Thr Val Gln Phe Val Cys Arg
Ala Asp Gly Asp Pro Pro Pro Ala 405 410 415 Ile Leu Trp Leu Ser Pro
Arg Lys His Leu Val Ser Ala Lys Ser Asn 420 425 430 Gly Arg Leu Thr
Val Phe Pro Asp Gly Thr Leu Glu Val Arg Tyr Ala 435 440 445 Gln Val
Gln Asp Asn Gly Thr Tyr Leu Cys Ile Ala Ala Asn Ala Gly 450 455 460
Gly Asn Asp Ser Met Pro Ala His Leu His Val Arg Ser Tyr Ser Pro 465
470 475 480 Asp Trp Pro His Gln Pro Asn Lys Thr Phe Ala Phe Ile Ser
Asn Gln 485 490 495 Pro Gly Glu Gly Glu Ala Asn Ser Thr Arg Ala Thr
Val Pro Phe Pro 500 505 510 Phe Asp Ile Lys Thr 515 2517PRTMacaca
fascicularis 2Thr Gly Cys Pro Pro Arg Cys Glu Cys Ser Ala Gln Asp
Arg Ala Val 1 5 10 15 Leu Cys His Arg Lys Arg Phe Val Ala Val Pro
Glu Gly Ile Pro Thr 20 25 30 Glu Thr Arg Leu Leu Asp Leu Gly Lys
Asn Arg Ile Lys Thr Leu Asn 35 40 45 Gln Asp Glu Phe Ala Ser Phe
Pro His Leu Glu Glu Leu Glu Leu Asn 50 55 60 Glu Asn Ile Val Ser
Ala Val Glu Pro Gly Ala Phe Asn Asn Leu Phe 65 70 75 80 Asn Leu Arg
Thr Leu Gly Leu Arg Ser Asn Arg Leu Lys Leu Ile Pro 85 90 95 Leu
Gly Val Phe Thr Gly Leu Ser Asn Leu Thr Lys Leu Asp Ile Ser 100 105
110 Glu Asn Lys Ile Val Ile Leu Leu Asp Tyr Met Phe Gln Asp Leu Tyr
115 120 125 Asn Leu Lys Ser Leu Glu Val Gly Asp Asn Asp Leu Val Tyr
Ile Ser 130 135 140 His Arg Ala Phe Ser Gly Leu Asn Ser Leu Glu Gln
Leu Thr Leu Glu 145 150 155 160 Lys Cys Asn Leu Thr Ser Ile Pro Thr
Glu Ala Leu Ser His Leu His 165 170 175 Gly Leu Ile Val Leu Arg Leu
Arg His Leu Asn Ile Asn Ala Ile Arg 180 185 190 Asp Tyr Ser Phe Lys
Arg Leu Tyr Arg Leu Lys Val Leu Glu Ile Ser 195 200 205 His Trp Pro
Tyr Leu Asp Thr Met Thr Pro Asn Cys Leu Tyr Gly Leu 210 215 220 Asn
Leu Thr Ser Leu Ser Ile Thr His Cys Asn Leu Thr Ala Val Pro 225 230
235 240 Tyr Leu Ala Val Arg His Leu Val Tyr Leu Arg Phe Leu Asn Leu
Ser 245 250 255 Tyr Asn Pro Ile Ser Thr Ile Glu Gly Ser Met Leu His
Glu Leu Leu 260 265 270 Arg Leu Gln Glu Ile Gln Leu Val Gly Gly Gln
Leu Ala Met Val Glu 275 280 285 Pro Tyr Ala Phe Arg Gly Leu Asn Tyr
Leu Arg Val Leu Asn Val Ser 290 295 300 Gly Asn Gln Leu Thr Thr Leu
Glu Glu Ser Val Phe His Ser Val Gly 305 310 315 320 Asn Leu Glu Thr
Leu Ile Leu Asp Ser Asn Pro Leu Ala Cys Asp Cys 325 330 335 Arg Leu
Leu Trp Val Phe Arg Arg Arg Trp Arg Leu Asn Phe Asn Arg 340 345 350
Gln Gln Pro Thr Cys Ala Thr Pro Glu Phe Val Gln Gly Lys Glu Phe 355
360 365 Lys Asp Phe Pro Asp Val Leu Leu Pro Asn Tyr Phe Thr Cys Arg
Arg 370 375 380 Ala Arg Ile Arg Asp Arg Lys Ala Gln Gln Val Phe Val
Asp Glu Gly 385 390 395 400 His Thr Val Gln Phe Val Cys Arg Ala Asp
Gly Asp Pro Pro Pro Ala 405 410 415 Ile Leu Trp Leu Ser Pro Arg Lys
His Leu Val Ser Ala Lys Ser Asn 420 425 430 Gly Arg Leu Thr Val Phe
Pro Asp Gly Thr Leu Glu Val Arg Tyr Ala 435 440 445 Gln Val Gln Asp
Asn Gly Thr Tyr Leu Cys Ile Ala Ala Asn Ala Gly 450 455 460 Gly Asn
Asp Ser Met Pro Ala His Leu His Val Arg Ser Tyr Ser Pro 465 470 475
480 Asp Trp Pro His Gln Pro Asn Lys Thr Phe Ala Phe Ile Pro Asn Gln
485 490 495 Pro Gly Glu Gly Glu Ala Asn Ser Thr Arg Ala Thr Val Pro
Phe Pro 500 505 510 Phe Asp Ile Lys Thr 515 3517PRTHomo sapiens
3Thr Gly Cys Pro Pro Arg Cys Glu Cys Ser Ala Gln Asp Arg Ala Val 1
5 10 15 Leu Cys His Arg Lys Arg Phe Val Ala Val Pro Glu Gly Ile Pro
Thr 20 25 30 Glu Thr Arg Leu Leu Asp Leu Gly Lys Asn Arg Ile Lys
Thr Leu Asn 35 40 45 Gln Asp Glu Phe Ala Ser Phe Pro His Leu Glu
Glu Leu Glu Leu Asn 50 55 60 Glu Asn Ile Val Ser Ala Val Glu Pro
Gly Ala Phe Asn Asn Leu Phe 65 70 75 80 Asn Leu Arg Thr Leu Gly Leu
Arg Ser Asn Arg Leu Lys Leu Ile Pro 85 90 95 Leu Gly Val Phe Thr
Gly Leu Ser Asn Leu Thr Lys Leu Asp Ile Ser 100 105 110 Glu Asn Lys
Ile Val Ile Leu Leu Asp Tyr Met Phe Gln Asp Leu Tyr 115 120 125 Asn
Leu Lys Ser Leu Glu Val Gly Asp Asn Asp Leu Val Tyr Ile Ser 130 135
140 His Arg Ala Phe Ser Gly Leu Asn Ser Leu Glu Gln Leu Thr Leu Glu
145 150 155 160 Lys Cys Asn Leu Thr Ser Ile Pro Thr Glu Ala Leu Ser
His Leu His 165 170 175 Gly Leu Ile Val Leu Arg Leu Arg His Leu Asn
Ile Asn Ala Ile Arg 180 185 190 Asp Tyr Ser Phe Lys Arg Leu Tyr Arg
Leu Lys Val Leu Glu Ile Ser 195 200 205 His Trp Pro Tyr Leu Asp Thr
Met Thr Pro Asn Cys Leu Tyr Gly Leu 210 215 220 Asn Leu Thr Ser Leu
Ser Ile Thr His Cys Asn Leu Thr Ala Val Pro 225 230 235 240 Tyr Leu
Ala Val Arg His Leu Val Tyr Leu Arg Phe Leu Asn Leu Ser 245 250 255
Tyr Asn Pro Ile Ser Thr Ile Glu Gly Ser Met Leu His Glu Leu Leu 260
265 270 Arg Leu Gln Glu Ile Gln Leu Val Gly Gly Gln Leu Ala Val Val
Glu 275 280 285 Pro Tyr Ala Phe Arg Gly Leu Asn Tyr Leu Arg Val Leu
Asn Val Ser 290 295 300 Gly Asn Gln Leu Thr Thr Leu Glu Glu Ser Val
Phe His Ser Val Gly 305 310 315 320 Asn Leu Glu Thr Leu Ile Leu Asp
Ser Asn Pro Leu Ala Cys Asp Cys 325 330 335 Arg Leu Leu Trp Val Phe
Arg Arg Arg Trp Arg Leu Asn Phe Asn Arg 340 345 350 Gln Gln Pro Thr
Cys Ala Thr Pro Glu Phe Val Gln Gly Lys Glu Phe 355 360 365 Lys Asp
Phe Pro Asp Val Leu Leu Pro Asn Tyr Phe Thr Cys Arg Arg 370 375 380
Ala Arg Ile Arg Asp Arg Lys Ala Gln Gln Val Phe Val Asp Glu Gly 385
390 395 400 His Thr Val Gln Phe Val Cys Arg Ala Asp Gly Asp Pro Pro
Pro Ala 405 410 415 Ile Leu Trp Leu Ser Pro Arg Lys His Leu Val Ser
Ala Lys Ser Asn 420 425 430 Gly Arg Leu Thr Val Phe Pro Asp Gly Thr
Leu Glu Val Arg Tyr Ala 435 440 445 Gln Val Gln Asp Asn Gly Thr Tyr
Leu Cys Ile Ala Ala Asn Ala Gly 450 455 460 Gly Asn Asp Ser Met Pro
Ala His Leu His Val Arg Ser Tyr Ser Pro 465 470 475 480 Asp Trp Pro
His Gln Pro Asn Lys Thr Phe Ala Phe Ile Ser Asn Gln 485 490 495 Pro
Gly Glu Gly Glu Ala Asn Ser Thr Arg Ala Thr Val Pro Phe Pro 500 505
510 Phe Asp Ile Lys Thr 515 4108PRTartificial4784 VL 4Asp Ile Glu
Leu Thr Gln Pro Pro Ser Val Ser Val Ala Pro Gly Gln 1 5 10 15 Thr
Ala Arg Ile Ser Cys Ser Gly Asp Asn Ile Gly Asn Tyr Tyr Val 20 25
30 Tyr Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Val Leu Val Ile Tyr
35 40 45 Glu Asp Thr Asn Arg Pro Ser Gly Ile Pro Glu Arg Phe Ser
Gly Ser 50 55 60 Asn Ser Gly Asn Thr Ala Thr Leu Thr Ile Ser Gly
Thr Gln Ala Glu 65 70 75 80 Asp Glu Ala Asp Tyr Tyr Cys Gln Ser Tyr
Asp Asn Leu His Glu Gln 85 90 95 Val Phe Gly Gly Gly Thr Lys Leu
Thr Val Leu Gly 100 105 5124PRTArtificial4784 VH 5Gln Val Gln Leu
Lys Glu Ser Gly Pro Ala Leu Val Lys Pro Thr Gln 1 5 10 15 Thr Leu
Thr Leu Thr Cys Thr Phe Ser Gly Phe Ser Leu Ser Ser Ser 20 25 30
Gly Val Gly Val Gly Trp Ile Arg Gln Pro Pro Gly Lys Ala Leu Glu 35
40 45 Trp Leu Ala His Ile Gly Ser Asp Asp Asp Lys Tyr Tyr Ser Thr
Ser 50 55 60 Leu Lys Thr Arg Leu Thr Ile Ser Lys Asp Thr Ser Lys
Asn Gln Val 65 70 75 80 Val Leu Thr Met Thr Asn Met Asp Pro Val Asp
Thr Ala Thr Tyr Tyr 85 90 95 Cys Ala Arg Asn Gln Gln Tyr Gly Asp
Gly Tyr Pro Gly Tyr Phe Asp 100 105 110 Tyr Trp Gly Gln Gly Thr Leu
Val Thr Val Ser Ser 115 120 6109PRTArtificial4785 VL 6Asp Ile Val
Leu Thr Gln Pro Pro Ser Val Ser Gly Ala Pro Gly Gln 1 5 10 15 Arg
Val Thr Ile Ser Cys Ser Gly Ser Ser Ser Asn Ile Gly Asn Asn 20 25
30 Tyr Val Ser Trp Tyr Gln Gln Leu Pro Gly Thr Ala Pro Lys Leu Leu
35 40 45 Ile Ser Arg Asn Ser Lys Arg Pro Ser Gly Val Pro Asp Arg
Phe Ser 50 55 60 Gly Ser Lys Ser Gly Thr Ser Ala Ser Leu Ala Ile
Thr Gly Leu Gln 65 70 75 80 Ser Glu Asp Glu Ala Asp Tyr Tyr Cys Ser
Thr Tyr Asp Thr Phe Ser 85 90 95 Ile Val Phe Gly Gly Gly Thr Lys
Leu Thr Val Leu Gly 100 105 7121PRTartificial4785 VH 7Gln Val Gln
Leu Gln Gln Ser Gly Pro Gly Leu Val Lys Pro Ser Gln 1 5 10 15 Thr
Leu Ser Leu Thr Cys Ala Ile Ser Gly Asp Ser Val Ser Asp Asn 20 25
30 Ser Ala Ala Trp Ser Trp Ile Arg Gln Ser Pro Gly Arg Gly Leu Glu
35 40 45 Trp Leu Gly Leu Ile Tyr Leu Arg Ser Lys Trp Asp Asn Asp
Tyr Ala 50 55 60 Val Ser Val Lys Ser Arg Ile Thr Ile Asn Pro Asp
Thr Ser Lys Asn 65 70 75 80 Gln Phe Ser Leu Gln Leu Asn Ser Val Thr
Pro Glu Asp Thr Ala Val 85 90 95 Tyr Tyr Cys Ala Arg Thr Gly Arg
Ala Asp Glu Phe Asp Val Trp Gly 100 105 110 Gln Gly Thr Leu Val Thr
Val Ser Ser 115 120 8372DNAartificialDNA 4784 VH 8caggtgcaat
tgaaagaaag cggcccggcc ctggtgaaac cgacccaaac cctgaccctg 60acctgtacct
tttccggatt tagcctgtct tcttctggtg ttggtgtggg ttggattcgc
120cagccgcctg ggaaagccct cgagtggctg gctcatatcg gttctgatga
tgataagtat 180tatagcacca gcctgaaaac gcgtctgacc attagcaaag
atacttcgaa aaatcaggtg 240gtgctgacta tgaccaacat ggacccggtg
gatacggcca cctattattg cgcgcgtaat 300cagcagtatg gtgatggtta
tcctggttat tttgattatt ggggccaagg caccctggtg 360acggttagct ca
3729363DNAartificialDNA 4785 VH 9caggtgcaat tgcaacagtc tggtccgggc
ctggtgaaac cgagccaaac cctgagcctg 60acctgtgcga tttccggaga tagcgtgagc
gataattctg ctgcttggtc ttggattcgc 120cagtctcctg ggcgtggcct
cgagtggctg ggccttatct atcttcgtag caagtgggat 180aacgattatg
cggtgagcgt gaaaagccgg attaccatca acccggatac ttcgaaaaac
240cagtttagcc tgcaactgaa cagcgtgacc ccggaagata cggccgtgta
ttattgcgcg 300cgtactggtc gtgctgatga gtttgatgtt tggggccaag
gcaccctggt gacggttagc 360tca 36310327DNAartificialDNA 4784 VL
10gatatcgaac tgacccagcc gccttcagtg agcgttgcac caggtcagac cgcgcgtatc
60tcgtgtagcg gcgataatat tggtaattat tatgtttatt ggtaccagca gaaacccggg
120caggcgccag ttcttgtgat ttatgaggat actaatcgtc cctcaggcat
cccggaacgc 180tttagcggat ccaacagcgg caacaccgcg accctgacca
ttagcggcac tcaggcggaa 240gacgaagcgg attattattg ccagtcttat
gataatcttc atgagcaggt gtttggcggc 300ggcacgaagt taaccgttct tggccag
32711330DNAartificialDNA 4785 VL 11gatatcgtgc tgacccagcc gccttcagtg
agtggcgcac caggtcagcg tgtgaccatc 60tcgtgtagcg gcagcagcag caacattggt
aataattatg tgtcttggta ccagcagttg 120cccgggacgg cgccgaaact
tctgatttct cgtaattcta agcgtccctc aggcgtgccg 180gatcgtttta
gcggatccaa aagcggcacc agcgcgagcc ttgcgattac gggcctgcaa
240agcgaagacg aagcggatta ttattgctct acttatgata ctttttctat
tgtgtttggc 300ggcggcacga agttaaccgt tcttggccag
330127PRTartificialAntibody 4784 CDR-H1 12Ser Ser Gly Val Gly Val
Gly 1
5 1316PRTartificialAntibody 4784 CDR-H2 13His Ile Gly Ser Asp Asp
Asp Lys Tyr Tyr Ser Thr Ser Leu Lys Thr 1 5 10 15
1414PRTartificialAntibody 4784 CDR-H3 14Asn Gln Gln Tyr Gly Asp Gly
Tyr Pro Gly Tyr Phe Asp Tyr 1 5 10 1511PRTartificialAntibody 4784
CDR-L1 15Ser Gly Asp Asn Ile Gly Asn Tyr Tyr Val Tyr 1 5 10
167PRTartificialAntibody 4784 CDR-L2 16Glu Asp Thr Asn Arg Pro Ser
1 5 1710PRTartificialAntibody 4784 CDR-L3 17Gln Ser Tyr Asp Asn Leu
His Glu Gln Val 1 5 10 187PRTartificialAntibody 4785 CDR'-H1 18Asp
Asn Ser Ala Ala Trp Ser 1 5 1918PRTartificialAntibody 4785 CDR'-H2
19Leu Ile Tyr Leu Arg Ser Lys Trp Asp Asn Asp Tyr Ala Val Ser Val 1
5 10 15 Lys Ser 209PRTartificialAntibody 4785 CDR'-H3 20Thr Gly Arg
Ala Asp Glu Phe Asp Val 1 5 2113PRTartificialAntibody 4785 CDR'-L1
21Ser Gly Ser Ser Ser Asn Ile Gly Asn Asn Tyr Val Ser 1 5 10
227PRTartificialAntibody 4785 CDR'-L2 22Arg Asn Ser Lys Arg Pro Ser
1 5 239PRTartificialAntibody 4785 CDR'-L3 23Ser Thr Tyr Asp Thr Phe
Ser Ile Val 1 5 2428DNAartificialDM22 24ggttatctcg agaccggctg
cccgcccc 282537DNAartificialDM23 25ggcccttcta gatcactcgc ctggctggtt
ggagatg 37266602DNAartificialAPtag-5-NHIS vector 26gacggatcgg
gagatctccc gatcccctat ggtcgactct cagtacaatc tgctctgatg 60ccgcatagtt
aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
120cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg
aagaatctgc 180ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc
cagatatacg cgttgacatt 240gattattgac tagttattaa tagtaatcaa
ttacggggtc attagttcat agcccatata 300tggagttccg cgttacataa
cttacggtaa atggcccgcc tggctgaccg cccaacgacc 360cccgcccatt
gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
420attgacgtca atgggtggac tatttacggt aaactgccca cttggcagta
catcaagtgt 480atcatatgcc aagtacgccc cctattgacg tcaatgacgg
taaatggccc gcctggcatt 540atgcccagta catgacctta tgggactttc
ctacttggca gtacatctac gtattagtca 600tcgctattac catggtgatg
cggttttggc agtacatcaa tgggcgtgga tagcggtttg 660actcacgggg
atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
720aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg
caaatgggcg 780gtaggcgtgt acggtgggag gtctatataa gcagagctct
ctggctaact agagaaccca 840ctgcttactg gcttatcgaa attaatacga
ctcactatag ggagacccaa gctggctagc 900caccatggag acagacacac
tcctgctatg ggtactgctg ctctgggttc caggttccac 960tggtgacgcg
gcccagccgg cccatcatca tcatcatcat gaagcttacg taagatcttc
1020cggaatcatc ccagttgagg aggagaaccc ggacttctgg aaccgcgagg
cagccgaggc 1080cctgggtgcc gccaagaagc tgcagcctgc acagacagcc
gccaagaacc tcatcatctt 1140cctgggcgat gggatggggg tgtctacggt
gacagctgcc aggatcctaa aagggcagaa 1200gaaggacaaa ctggggcctg
agatacccct ggccatggac cgcttcccat atgtggctct 1260gtccaagaca
tacaatgtag acaaacatgt gccagacagt ggagccacag ccacggccta
1320cctgtgcggg gtcaagggca acttccagac cattggcttg agtgcagccg
cccgctttaa 1380ccagtgcaac acgacacgcg gcaacgaggt catctccgtg
atgaatcggg ccaagaaagc 1440agggaagtca gtgggagtgg taaccaccac
acgagtgcag cacgcctcgc cagccggcac 1500ctacgcccac acggtgaacc
gcaactggta ctcggacgcc gacgtgcctg cctcggcccg 1560ccaggagggg
tgccaggaca tcgctacgca gctcatctcc aacatggaca ttgacgtgat
1620cctaggtgga ggccgaaagt acatgtttcg catgggaacc ccagaccctg
agtacccaga 1680tgactacagc caaggtggga ccaggctgga cgggaagaat
ctggtgcagg aatggctggc 1740gaagcgccag ggtgcccggt atgtgtggaa
ccgcactgag ctcatgcagg cttccctgga 1800cccgtctgtg acccatctca
tgggtctctt tgagcctgga gacatgaaat acgagatcca 1860ccgagactcc
acactggacc cctccctgat ggagatgaca gaggctgccc tgcgcctgct
1920gagcaggaac ccccgcggct tcttcctctt cgtggagggt ggtcgcatcg
accatggtca 1980tcatgaaagc agggcttacc gggcactgac tgagacgatc
atgttcgacg acgccattga 2040gagggcgggc cagctcacca gcgaggagga
cacgctgagc ctcgtcactg ccgaccactc 2100ccacgtcttc tccttcggag
gctaccccct gcgagggagc tccatcttcg ggctggcccc 2160tggcaaggcc
cgggacagga aggcctacac ggtcctccta tacggaaacg gtccaggcta
2220tgtgctcaag gacggcgccc ggccggatgt taccgagagc gagagcggga
gccccgagta 2280tcggcagcag tcagcagtgc ccctggacga agagacccac
gcaggcgagg acgtggcggt 2340gttcgcgcgc ggcccgcagg cgcacctggt
tcacggcgtg caggagcaga ccttcatagc 2400gcacgtcatg gccttcgccg
cctgcctgga gccctacacc gcctgcgacc tggcgccccc 2460cgccggcacc
accgacgccg cgcacccggg ttatctcgag gaagcgctct ctctagaagg
2520gcccgaacaa aaactcatct cagaagagga tctgaatagc gccgtcgacc
atcatcatca 2580tcatcattga gtttaaaccc gctgatcagc ctcgactgtg
ccttctagtt gccagccatc 2640tgttgtttgc ccctcccccg tgccttcctt
gaccctggaa ggtgccactc ccactgtcct 2700ttcctaataa aatgaggaaa
ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg 2760gggtggggtg
gggcaggaca gcaaggggga ggattgggaa gacaatagca ggcatgctgg
2820ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc agctggggct
ctagggggta 2880tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt
gtggtggtta cgcgcagcgt 2940gaccgctaca cttgccagcg ccctagcgcc
cgctcctttc gctttcttcc cttcctttct 3000cgccacgttc gccggctttc
cccgtcaagc tctaaatcgg ggcatccctt tagggttccg 3060atttagtgct
ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag
3120tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca
cgttctttaa 3180tagtggactc ttgttccaaa ctggaacaac actcaaccct
atctcggtct attcttttga 3240tttataaggg attttgggga tttcggccta
ttggttaaaa aatgagctga tttaacaaaa 3300atttaacgcg aattaattct
gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc 3360tccccagcag
gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga
3420aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
ttagtcagca 3480accatagtcc cgcccctaac tccgcccatc ccgcccctaa
ctccgcccag ttccgcccat 3540tctccgcccc atggctgact aatttttttt
atttatgcag aggccgaggc cgcctctgcc 3600tctgagctat tccagaagta
gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag 3660ctcccgggag
cttgtatatc cattttcgga tctgatcagc acgtgttgac aattaatcat
3720cggcatagta tatcggcata gtataatacg acaaggtgag gaactaaacc
atggccaagt 3780tgaccagtgc cgttccggtg ctcaccgcgc gcgacgtcgc
cggagcggtc gagttctgga 3840ccgaccggct cgggttctcc cgggacttcg
tggaggacga cttcgccggt gtggtccggg 3900acgacgtgac cctgttcatc
agcgcggtcc aggaccaggt ggtgccggac aacaccctgg 3960cctgggtgtg
ggtgcgcggc ctggacgagc tgtacgccga gtggtcggag gtcgtgtcca
4020cgaacttccg ggacgcctcc gggccggcca tgaccgagat cggcgagcag
ccgtgggggc 4080gggagttcgc cctgcgcgac ccggccggca actgcgtgca
cttcgtggcc gaggagcagg 4140actgacacgt gctacgagat ttcgattcca
ccgccgcctt ctatgaaagg ttgggcttcg 4200gaatcgtttt ccgggacgcc
ggctggatga tcctccagcg cggggatctc atgctggagt 4260tcttcgccca
ccccaacttg tttattgcag cttataatgg ttacaaataa agcaatagca
4320tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt
ttgtccaaac 4380tcatcaatgt atcttatcat gtctgtatac cgtcgacctc
tagctagagc ttggcgtaat 4440catggtcata gctgtttcct gtgtgaaatt
gttatccgct cacaattcca cacaacatac 4500gagccggaag cataaagtgt
aaagcctggg gtgcctaatg agtgagctaa ctcacattaa 4560ttgcgttgcg
ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat
4620gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
gcttcctcgc 4680tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
ggtatcagct cactcaaagg 4740cggtaatacg gttatccaca gaatcagggg
ataacgcagg aaagaacatg tgagcaaaag 4800gccagcaaaa ggccaggaac
cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc 4860gcccccctga
cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
4920gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
cctgttccga 4980ccctgccgct taccggatac ctgtccgcct ttctcccttc
gggaagcgtg gcgctttctc 5040aatgctcacg ctgtaggtat ctcagttcgg
tgtaggtcgt tcgctccaag ctgggctgtg 5100tgcacgaacc ccccgttcag
cccgaccgct gcgccttatc cggtaactat cgtcttgagt 5160ccaacccggt
aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
5220gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
tacggctaca 5280ctagaaggac agtatttggt atctgcgctc tgctgaagcc
agttaccttc ggaaaaagag 5340ttggtagctc ttgatccggc aaacaaacca
ccgctggtag cggtggtttt tttgtttgca 5400agcagcagat tacgcgcaga
aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg 5460ggtctgacgc
tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
5520aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
atctaaagta 5580tatatgagta aacttggtct gacagttacc aatgcttaat
cagtgaggca cctatctcag 5640cgatctgtct atttcgttca tccatagttg
cctgactccc cgtcgtgtag ataactacga 5700tacgggaggg cttaccatct
ggccccagtg ctgcaatgat accgcgagac ccacgctcac 5760cggctccaga
tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
5820ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
agagtaagta 5880gttcgccagt taatagtttg cgcaacgttg ttgccattgc
tacaggcatc gtggtgtcac 5940gctcgtcgtt tggtatggct tcattcagct
ccggttccca acgatcaagg cgagttacat 6000gatcccccat gttgtgcaaa
aaagcggtta gctccttcgg tcctccgatc gttgtcagaa 6060gtaagttggc
cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
6120tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
tcattctgag 6180aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
aatacgggat aataccgcgc 6240cacatagcag aactttaaaa gtgctcatca
ttggaaaacg ttcttcgggg cgaaaactct 6300caaggatctt accgctgttg
agatccagtt cgatgtaacc cactcgtgca cccaactgat 6360cttcagcatc
ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
6420ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
ttcctttttc 6480aatattattg aagcatttat cagggttatt gtctcatgag
cggatacata tttgaatgta 6540tttagaaaaa taaacaaata ggggttccgc
gcacatttcc ccgaaaagtg ccacctgacg 6600tc 6602271743DNAHomo sapiens
27acgggctgcc cgccccgctg cgagtgctcc gcccaggacc gcgctgtgct gtgccaccgc
60aagcgctttg tggcagtccc cgagggcatc cccaccgaga cgcgcctgct ggacctaggc
120aagaaccgca tcaaaacgct caaccaggac gagttcgcca gcttcccgca
cctggaggag 180ctggagctca acgagaacat cgtgagcgcc gtggagcccg
gcgccttcaa caacctcttc 240aacctccgga cgctgggtct ccgcagcaac
cgcctgaagc tcatcccgct aggcgtcttc 300actggcctca gcaacctgac
caagctggac atcagcgaga acaagattgt tatcctgctg 360gactacatgt
ttcaggacct gtacaacctc aagtcactgg aggttggcga caatgacctc
420gtctacatct ctcaccgcgc cttcagcggc ctcaacagcc tggagcagct
gacgctggag 480aaatgcaacc tgacctccat ccccaccgag gcgctgtccc
acctgcacgg cctcatcgtc 540ctgaggctcc ggcacctcaa catcaatgcc
atccgggact actccttcaa gaggctctac 600cgactcaagg tcttggagat
ctcccactgg ccctacttgg acaccatgac acccaactgc 660ctctacggcc
tcaacctgac gtccctgtcc atcacacact gcaatctgac cgctgtgccc
720tacctggccg tccgccacct agtctatctc cgcttcctca acctctccta
caaccccatc 780agcaccattg agggctccat gttgcatgag ctgctccggc
tgcaggagat ccagctggtg 840ggcgggcagc tggccgtggt ggagccctat
gccttccgcg gcctcaacta cctgcgcgtg 900ctcaatgtct ctggcaacca
gctgaccaca ctggaggaat cagtcttcca ctcggtgggc 960aacctggaga
cactcatcct ggactccaac ccgctggcct gcgactgtcg gctcctgtgg
1020gtgttccggc gccgctggcg gctcaacttc aaccggcagc agcccacgtg
cgccacgccc 1080gagtttgtcc agggcaagga gttcaaggac ttccctgatg
tgctactgcc caactacttc 1140acctgccgcc gcgcccgcat ccgggaccgc
aaggcccagc aggtgtttgt ggacgagggc 1200cacacggtgc agtttgtgtg
ccgggccgat ggcgacccgc cgcccgccat cctctggctc 1260tcaccccgaa
agcacctggt ctcagccaag agcaatgggc ggctcacagt cttccctgat
1320ggcacgctgg aggtgcgcta cgcccaggta caggacaacg gcacgtacct
gtgcatcgcg 1380gccaacgcgg gcggcaacga ctccatgccc gcccacctgc
atgtgcgcag ctactcgccc 1440gactggcccc atcagcccaa caagaccttc
gctttcatct ccaaccagcc gggcgaggga 1500gaggccaaca gcacccgcgc
cactgtgcct ttccccttcg acatcaagac cctcatcatc 1560gccaccacca
tgggcttcat ctctttcctg ggcgtcgtcc tcttctgcct ggtgctgctg
1620tttctctgga gccggggcaa gggcaacaca aagcacaaca tcgagatcga
gtatgtgccc 1680cgaaagtcgg acgcaggcat cagctccgcc gacgcgcccc
gcaagttcaa catgaagatg 1740ata 1743281743DNAMacaca fascicularis
28acgggctgcc cgccccgctg cgagtgctcc gcccaggacc gggctgtgct ctgccaccgc
60aagcgctttg tggcagtgcc tgagggcatc cccacggaga cgcgcctgct ggacctgggg
120aagaaccgca tcaaaacgct caaccaggac gagttcgcca gcttcccgca
cctggaggag 180ctggagctca acgagaacat cgtgagcgcc gtggagcctg
gcgccttcaa caaccttttc 240aacctccgga cgctgggtct ccgcagcaac
cgcctgaagc tcatcccgct gggcgtcttc 300actggcctca gcaacttgac
caagctggac atcagcgaga acaagatcgt tatcctgctg 360gactacatgt
tccaggacct gtacaacctc aagtcactgg aggttggcga caatgacctc
420gtctacatct cccaccgcgc cttcagcggc ctcaacagcc tggagcagct
gacgctggag 480aaatgcaacc tgacctccat ccccaccgag gcgctgtccc
acctgcacgg cctcatcgtc 540ctgaggctcc ggcacctcaa catcaatgcc
atccgggact actccttcaa gaggttgtac 600cgactcaagg tcttggagat
ctcccactgg ccctacttgg acaccatgac acccaactgc 660ctctacggcc
tcaacctgac gtccctgtcc atcacgcact gcaatctgac cgctgtgccc
720tacctggccg tccgccacct ggtctatctc cgcttcctca acctctccta
caaccccatc 780agcaccattg agggctccat gttgcatgag ctgctccggc
tgcaggagat ccagctggtg 840ggcgggcagc tggccatggt ggagccctat
gccttccgcg gcctcaacta cctgcgcgtg 900ctcaatgtct ctggcaacca
gctgaccacg ctggaagaat cagtcttcca ctcggtgggc 960aacctggaga
cgctcatcct ggactccaac ccactggcct gcgactgtcg gctcctgtgg
1020gtgttccggc gccgctggcg gctcaacttc aaccggcagc agcccacgtg
cgccacgccc 1080gagttcgtcc agggcaagga gttcaaggac ttccctgatg
tgctactgcc caactacttc 1140acctgccgcc gcgcccgcat ccgggatcgc
aaggcccagc aggtgtttgt ggatgagggc 1200cacacggtgc agtttgtgtg
ccgggccgat ggcgacccgc cgcccgccat cctctggctc 1260tcaccccgaa
agcacctggt ctcagccaag agcaatgggc ggctcacagt cttccctgat
1320ggcacgctgg aggtgcgcta cgcccaggta caggacaatg gcacgtacct
gtgcatcgcg 1380gccaatgcag gcggcaacga ctccatgcct gcccacctgc
atgtgcgcag ctactcaccc 1440gactggcccc atcagcccaa caagaccttc
gccttcatcc ccaaccagcc gggcgaggga 1500gaggccaaca gcacccgagc
cactgtgcct ttccccttcg acatcaagac cctcatcatc 1560gccaccacca
tgggcttcat ctctttcctg ggcgtcgtcc tcttctgcct ggtgctgctg
1620tttctctgga gccggggcaa gggcaacacg aagcacaaca tcgagatcga
gtatgtcccc 1680cgaaagtcgg acgcaggcat cagctccgcc gacgcgcccc
gcaagttcaa catgaagatg 1740ata 1743291743DNARattus sp. 29accggctgcc
cgccccgctg cgagtgctca gcgcaggacc gagcagtgct ctgtcaccgc 60aagcgctttg
tggcggtgcc cgagggcatc cccaccgaga ctcgcctgct ggacctgggc
120aaaaaccgca tcaagacact caaccaggac gagtttgcca gtttcccaca
cctggaggag 180ctagaactca atgagaacat tgtgagcgct gtggagccgg
gcgccttcaa caacctcttc 240aacctgagga cgctggggct tcgcagcaac
cgcctgaagc tcatcccgct gggcgtcttc 300accggcctca gcaacttgac
caagctggac atcagcgaga acaagatcgt catcctgcta 360gactacatgt
tccaagacct atacaacctc aagtcgctgg aggtcggcga caatgacctc
420gtctacatct cccatcgagc cttcagcggc ctcaacagcc tggaacagct
gacgctggag 480aaatgcaatc tgacctccat ccccactgag gcactctccc
acctgcatgg cctcatcgtc 540ctgcggctac gacacctcaa catcaatgcc
atacgggact actccttcaa gaggctgtac 600cgactcaagg tcttagagat
ctcccactgg ccctacctgg acaccatgac ccccaactgc 660ctctacggcc
tcaacctgac atccctatct atcacgcact gcaacctgac agccgtgccc
720tatctggcag tgcgccacct ggtctatctc cgtttcctca atctttccta
caaccccatc 780ggtacaatcg agggctccat gctgcatgag ctgctgcggt
tgcaagagat ccaactggtg 840ggcgggcagc tggccgtggt ggagccctac
gcctttcgtg ggctcaatta cctgcgtgtg 900ctcaatgttt ctggcaacca
gctgaccacc ctggaggagt cagccttcca ctcggtgggc 960aacctggaga
cgctcattct ggactccaac ccactggcct gtgactgccg gctgctgtgg
1020gtgttccggc gccgctggcg gctcaacttc aacaggcagc agcctacctg
cgccacacct 1080gagttcgtcc agggcaagga gttcaaggac ttccccgatg
tgctcctacc caactacttc 1140acctgccgcc gggcccacat ccgggaccgc
aaggcacagc aggtgtttgt agatgagggc 1200cacacggtgc agttcgtatg
ccgggcagat ggcgaccctc caccagctat cctttggctc 1260tcaccccgca
agcacttggt ctcagccaag agcaatgggc ggctcacagt cttccctgat
1320ggcacgctgg aggtgcgcta cgcccaggta caggacaacg gcacgtacct
gtgcatcgca 1380gccaatgcag gcggcaacga ctccatgccc gcccacttgc
atgtgcgcag ctactcgcct 1440gactggcccc atcaacccaa caagaccttc
gccttcatct ccaaccagcc aggcgaggga 1500gaggccaaca gcacccgcgc
cactgtgcct ttccccttcg acatcaagac gctcatcatc 1560gccaccacca
tgggcttcat ctccttcctg ggcgtggtcc tattctgcct ggtgctgctg
1620tttctatgga gccggggcaa aggcaacaca aagcacaaca tcgaaattga
atatgtgccc 1680cggaaatcgg acgcaggcat cagctcagct gatgcacccc
gcaagttcaa catgaagatg 1740ata 17433030DNAartificialDM14
30ctacgtctag aacgggctgc ccgccccgct 303130DNAartificialDM15
31ctacgtctag aacgggctgc ccgccccgct 30325045DNAartificialpSecTag2-V5
vector 32gacggatcgg gagatctccc gatcccctat ggtcgactct cagtacaatc
tgctctgatg 60ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct
gagtagtgcg 120cgagcaaaat ttaagctaca acaaggcaag gcttgaccga
caattgcatg aagaatctgc 180ttagggttag gcgttttgcg ctgcttcgcg
atgtacgggc cagatatacg cgttgacatt 240gattattgac tagttattaa
tagtaatcaa ttacggggtc attagttcat agcccatata 300tggagttccg
cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
360cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata
gggactttcc 420attgacgtca atgggtggac tatttacggt aaactgccca
cttggcagta catcaagtgt 480atcatatgcc aagtacgccc cctattgacg
tcaatgacgg taaatggccc gcctggcatt 540atgcccagta catgacctta
tgggactttc ctacttggca gtacatctac gtattagtca 600tcgctattac
catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
660actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg
ttttggcacc 720aaaatcaacg ggactttcca aaatgtcgta acaactccgc
cccattgacg caaatgggcg 780gtaggcgtgt acggtgggag gtctatataa
gcagagctct ctggctaact agagaaccca 840ctgcttactg gcttatcgaa
attaatacga ctcactatag ggagacccaa gctggctagc 900caccatggag
acagacacac tcctgctatg ggtactgctg ctctgggttc caggttccac
960tggtgacgcg gcccagcccg gtaagcctat ccctaaccct ctcctcggtc
tcgattctac 1020gtctagatat cctcgagaaa cccgctgatc agcctcgact
gtgccttcta gttgccagcc 1080atctgttgtt tgcccctccc ccgtgccttc
cttgaccctg gaaggtgcca ctcccactgt 1140cctttcctaa taaaatgagg
aaattgcatc gcattgtctg agtaggtgtc attctattct 1200ggggggtggg
gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatgc
1260tggggatgcg gtgggctcta
tggcttctga ggcggaaaga accagctggg gctctagggg 1320gtatccccac
gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag
1380cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct
tcccttcctt 1440tctcgccacg ttcgccggct ttccccgtca agctctaaat
cggggcatcc ctttagggtt 1500ccgatttagt gctttacggc acctcgaccc
caaaaaactt gattagggtg atggttcacg 1560tagtgggcca tcgccctgat
agacggtttt tcgccctttg acgttggagt ccacgttctt 1620taatagtgga
ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt
1680tgatttataa gggattttgg ggatttcggc ctattggtta aaaaatgagc
tgatttaaca 1740aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt
tagggtgtgg aaagtcccca 1800ggctccccag caggcagaag tatgcaaagc
atgcatctca attagtcagc aaccaggtgt 1860ggaaagtccc caggctcccc
agcaggcaga agtatgcaaa gcatgcatct caattagtca 1920gcaaccatag
tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc
1980cattctccgc cccatggctg actaattttt tttatttatg cagaggccga
ggccgcctct 2040gcctctgagc tattccagaa gtagtgagga ggcttttttg
gaggcctagg cttttgcaaa 2100aagctcccgg gagcttgtat atccattttc
ggatctgatc agcacgtgtt gacaattaat 2160catcggcata gtatatcggc
atagtataat acgacaaggt gaggaactaa accatggcca 2220agttgaccag
tgccgttccg gtgctcaccg cgcgcgacgt cgccggagcg gtcgagttct
2280ggaccgaccg gctcgggttc tcccgggact tcgtggagga cgacttcgcc
ggtgtggtcc 2340gggacgacgt gaccctgttc atcagcgcgg tccaggacca
ggtggtgccg gacaacaccc 2400tggcctgggt gtgggtgcgc ggcctggacg
agctgtacgc cgagtggtcg gaggtcgtgt 2460ccacgaactt ccgggacgcc
tccgggccgg ccatgaccga gatcggcgag cagccgtggg 2520ggcgggagtt
cgccctgcgc gacccggccg gcaactgcgt gcacttcgtg gccgaggagc
2580aggactgaca cgtgctacga gatttcgatt ccaccgccgc cttctatgaa
aggttgggct 2640tcggaatcgt tttccgggac gccggctgga tgatcctcca
gcgcggggat ctcatgctgg 2700agttcttcgc ccaccccaac ttgtttattg
cagcttataa tggttacaaa taaagcaata 2760gcatcacaaa tttcacaaat
aaagcatttt tttcactgca ttctagttgt ggtttgtcca 2820aactcatcaa
tgtatcttat catgtctgta taccgtcgac ctctagctag agcttggcgt
2880aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt
ccacacaaca 2940tacgagccgg aagcataaag tgtaaagcct ggggtgccta
atgagtgagc taactcacat 3000taattgcgtt gcgctcactg cccgctttcc
agtcgggaaa cctgtcgtgc cagctgcatt 3060aatgaatcgg ccaacgcgcg
gggagaggcg gtttgcgtat tgggcgctct tccgcttcct 3120cgctcactga
ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
3180aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
atgtgagcaa 3240aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
gctggcgttt ttccataggc 3300tccgcccccc tgacgagcat cacaaaaatc
gacgctcaag tcagaggtgg cgaaacccga 3360caggactata aagataccag
gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc 3420cgaccctgcc
gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
3480ctcaatgctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
aagctgggct 3540gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
atccggtaac tatcgtcttg 3600agtccaaccc ggtaagacac gacttatcgc
cactggcagc agccactggt aacaggatta 3660gcagagcgag gtatgtaggc
ggtgctacag agttcttgaa gtggtggcct aactacggct 3720acactagaag
gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
3780gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
ttttttgttt 3840gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
agatcctttg atcttttcta 3900cggggtctga cgctcagtgg aacgaaaact
cacgttaagg gattttggtc atgagattat 3960caaaaaggat cttcacctag
atccttttaa attaaaaatg aagttttaaa tcaatctaaa 4020gtatatatga
gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
4080cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
tagataacta 4140cgatacggga gggcttacca tctggcccca gtgctgcaat
gataccgcga gacccacgct 4200caccggctcc agatttatca gcaataaacc
agccagccgg aagggccgag cgcagaagtg 4260gtcctgcaac tttatccgcc
tccatccagt ctattaattg ttgccgggaa gctagagtaa 4320gtagttcgcc
agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
4380cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca
aggcgagtta 4440catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
cggtcctccg atcgttgtca 4500gaagtaagtt ggccgcagtg ttatcactca
tggttatggc agcactgcat aattctctta 4560ctgtcatgcc atccgtaaga
tgcttttctg tgactggtga gtactcaacc aagtcattct 4620gagaatagtg
tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
4680cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg
gggcgaaaac 4740tctcaaggat cttaccgctg ttgagatcca gttcgatgta
acccactcgt gcacccaact 4800gatcttcagc atcttttact ttcaccagcg
tttctgggtg agcaaaaaca ggaaggcaaa 4860atgccgcaaa aaagggaata
agggcgacac ggaaatgttg aatactcata ctcttccttt 4920ttcaatatta
ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
4980gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa
gtgccacctg 5040acgtc 5045331743DNAHomo sapiens 33attggctgcc
ccgctcgctg tgagtgctct gcccagaaca aatctgttag ctgtcacaga 60aggcgattga
tcgccatccc agagggcatt cccatcgaaa ccaaaatctt ggacctcagt
120aaaaacaggc taaaaagcgt caaccctgaa gaattcatat catatcctct
gctggaagag 180atagacttga gtgacaacat cattgccaat gtggaaccag
gagcattcaa caatctcttt 240aacctgcgtt ccctccgcct aaaaggcaat
cgtctaaagc tggtcccttt gggagtattc 300acggggctgt ccaatctcac
taagcttgac attagtgaga ataagattgt cattttacta 360gactacatgt
tccaagatct acataacctg aagtctctag aagtggggga caatgatttg
420gtttatatat cacacagggc attcagtggg cttcttagct tggagcagct
caccctggag 480aaatgcaact taacagcagt accaacagaa gccctctccc
acctccgcag cctcatcagc 540ctgcatctga agcatctcaa tatcaacaat
atgcctgtgt atgcctttaa aagattgttc 600cacctgaaac acctagagat
tgactattgg cctttactgg atatgatgcc tgccaatagc 660ctctacggtc
tcaacctcac atccctttca gtcaccaaca ccaatctgtc tactgtaccc
720ttccttgcct ttaaacacct ggtatacctg actcacctta acctctccta
caatcccatc 780agcactattg aagcaggcat gttctctgac ctgatccgcc
ttcaggagct tcatatagtg 840ggggcccagc ttcgcaccat tgagcctcac
tccttccaag ggctccgctt cctacgcgtg 900ctcaatgtgt ctcagaacct
gctggaaact ttggaagaga atgtcttctc ctcccctagg 960gctctggagg
tcttgagcat taacaacaac cctctggcct gtgactgccg ccttctctgg
1020atcttgcagc gacagcccac cctgcagttt ggtggccagc aacctatgtg
tgctggccca 1080gacaccatcc gtgagaggtc tttcaaggat ttccatagca
ctgccctttc tttttacttt 1140acctgcaaaa aacccaaaat ccgtgaaaag
aagttgcagc atctgctagt agatgaaggg 1200cagacagtcc agctagaatg
cagtgcagat ggagacccgc agcctgtgat ttcctgggtg 1260acaccccgaa
ggcgtttcat caccaccaag tccaatggaa gagccaccgt gttgggtgat
1320ggcaccttgg aaatccgctt tgcccaggat caagacagcg ggatgtatgt
ttgcatcgct 1380agcaatgctg ctgggaatga taccttcaca gcctccttaa
ctgtgaaagg attcgcttca 1440gatcgttttc tttatgcgaa caggacccct
atgtacatga ccgactccaa tgacaccatt 1500tccaatggca ccaatgccaa
tactttttcc ctggacctta aaacaatact ggtgtctaca 1560gctatgggct
gcttcacatt cctgggagtg gttttatttt gttttcttct cctttttgtg
1620tggagccgag ggaaaggcaa gcacaaaaac agcattgacc ttgagtatgt
gcccagaaaa 1680aacaatggtg ctgttgtgga aggggaggta gctggaccca
ggaggttcaa catgaaaatg 1740att 17433430DNAartificialDM16
34ctacgtctag aattggctgc cccgctcgct 303539DNAartificialDM17
35ggtttctcga gtcaaatcat tttcatgttg aacctcctg
39367086DNAartificialpRS5a-IgG 36tcgacggatc gggagatccg ggacatgtac
ctcccagggg cccaggaaga ctacgggagg 60ctacaccaac gtcaatcaga ggggcctgtg
tagctaccga taagcggacc ctcaagaggg 120cattagcaat agtgtttata
aggccccctt gttaacccta aacgggtagc atatgcttcc 180cgggtagtag
tatatactat ccagactaac cctaattcaa tagcatatgt tacccaacgg
240gaagcatatg ctatcgaatt agggttagta aaagggtcct aaggaacagc
gatatctccc 300accccatgag ctgtcacggt tttatttaca tggggtcagg
attccacgag ggtagtgaac 360cattttagtc acaagggcag tggctgaaga
tcaaggagcg ggcagtgaac tctcctgaat 420cttcgcctgc ttcttcattc
tccttcgttt agctaataga ataactgctg agttgtgaac 480agtaaggtgt
atgtgaggtg ctcgaaaaca aggtttcagg tgacgccccc agaataaaat
540ttggacgggg ggttcagtgg tggcattgtg ctatgacacc aatataaccc
tcacaaaccc 600cttgggcaat aaatactagt gtaggaatga aacattctga
atatctttaa caatagaaat 660ccatggggtg gggacaagcc gtaaagactg
gatgtccatc tcacacgaat ttatggctat 720gggcaacaca taatcctagt
gcaatatgat actggggtta ttaagatgtg tcccaggcag 780ggaccaagac
aggtgaacca tgttgttaca ctctatttgt aacaagggga aagagagtgg
840acgccgacag cagcggactc cactggttgt ctctaacacc cccgaaaatt
aaacggggct 900ccacgccaat ggggcccata aacaaagaca agtggccact
cttttttttg aaattgtgga 960gtgggggcac gcgtcagccc ccacacgccg
ccctgcggtt ttggactgta aaataagggt 1020gtaataactt ggctgattgt
aaccccgcta accactgcgg tcaaaccact tgcccacaaa 1080accactaatg
gcaccccggg gaatacctgc ataagtaggt gggcgggcca agataggggc
1140gcgattgctg cgatctggag gacaaattac acacacttgc gcctgagcgc
caagcacagg 1200gttgttggtc ctcatattca cgaggtcgct gagagcacgg
tgggctaatg ttgccatggg 1260tagcatatac tacccaaata tctggatagc
atatgctatc ctaatctata tctgggtagc 1320ataggctatc ctaatctata
tctgggtagc atatgctatc ctaatctata tctgggtagt 1380atatgctatc
ctaatttata tctgggtagc ataggctatc ctaatctata tctgggtagc
1440atatgctatc ctaatctata tctgggtagt atatgctatc ctaatctgta
tccgggtagc 1500atatgctatc ctaatagaga ttagggtagt atatgctatc
ctaatttata tctgggtagc 1560atatactacc caaatatctg gatagcatat
gctatcctaa tctatatctg ggtagcatat 1620gctatcctaa tctatatctg
ggtagcatag gctatcctaa tctatatctg ggtagcatat 1680gctatcctaa
tctatatctg ggtagtatat gctatcctaa tttatatctg ggtagcatag
1740gctatcctaa tctatatctg ggtagcatat gctatcctaa tctatatctg
ggtagtatat 1800gctatcctaa tctgtatccg ggtagcatat gctatcctca
tgcatataca gtcagcatat 1860gatacccagt agtagagtgg gagtgctatc
ctttgcatat gccgccacct cccaaggggg 1920cgtgaatttt cgctgcttgt
ccttttcctg catgcggatc ttcaatattg gccattagcc 1980atattattca
ttggttatat agcataaatc aatattggct attggccatt gcatacgttg
2040tatctatatc ataatatgta catttatatt ggctcatgtc caatatgacc
gccatgttgg 2100cattgattat tgactagtta ttaatagtaa tcaattacgg
ggtcattagt tcatagccca 2160tatatggagt tccgcgttac ataacttacg
gtaaatggcc cgcctggctg accgcccaac 2220gacccccgcc cattgacgtc
aataatgacg tatgttccca tagtaacgcc aatagggact 2280ttccattgac
gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa
2340gtgtatcata tgccaagtcc gccccctatt gacgtcaatg acggtaaatg
gcccgcctgg 2400cattatgccc agtacatgac cttacgggac tttcctactt
ggcagtacat ctacgtatta 2460gtcatcgcta ttaccatggt gatgcggttt
tggcagtaca ccaatgggcg tggatagcgg 2520tttgactcac ggggatttcc
aagtctccac cccattgacg tcaatgggag tttgttttgg 2580caccaaaatc
aacgggactt tccaaaatgt cgtaataacc ccgccccgtt gacgcaaatg
2640ggcggtaggc gtgtacggtg ggaggtctat ataagcagag ctcgtttagt
gaaccgtcag 2700atcgcctgga gacgccatcc acgctgtttt gacctccata
gaagacaccg ggaccgatcc 2760agcctccgcg gccgggaacg gtgcattgga
acgcggattc cccgtgccaa gagtgacgta 2820agtaccgcct atagagtcta
taggcccacc cccttggctt cgttagaacg cggctacaat 2880taatacataa
ccttatgtat catacacata cgatttaggt gacactatag aataacatcc
2940actttgcctt tctctccaca ggtgtccact cccaggtcca actgcacgga
agcttcaatt 3000gggatccctc gaggttctgt tccagggtcc gaaatcttgt
gacaaaactc acacatgccc 3060accgtgccca gcacctgaac tcctgggggg
accgtcagtc ttcctcttcc ccccaaaacc 3120caaggacacc ctcatgatct
cccggacccc tgaggtcaca tgcgtggtgg tggacgtgag 3180ccacgaagac
cctgaggtca agttcaactg gtacgtggac ggcgtggagg tgcataatgc
3240caagacaaag ccgcgggagg agcagtacaa cagcacgtac cgtgtggtca
gcgtcctcac 3300cgtcctgcac caggactggc tgaatggcaa ggagtacaag
tgcaaggtct ccaacaaagc 3360cctcccagcc cccatcgaga aaaccatctc
caaagccaaa gggcagcccc gagaaccaca 3420ggtgtacacc ctgcccccat
cccgggatga gctgaccaag aaccaggtca gcctgacctg 3480cctggtcaaa
ggcttctatc ccagcgacat cgccgtggag tgggagagca atgggcagcc
3540ggagaacaac tacaagacca cgcctcccgt gctggactcc gacggctcct
tcttcctcta 3600cagcaagctc accgtggaca agagcaggtg gcagcagggg
aacgtcttct catgctccgt 3660gatgcatgag gctctgcaca accactacac
gcagaagagc ctctccctgt ctccgggtaa 3720atgagatctg gtacctcgcg
atggcggccg ctctagaggg cccgtttaaa cccgctgatc 3780agcctcgact
gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc
3840cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg
aaattgcatc 3900gcattgtctg agtaggtgtc attctattct ggggggtggg
gtggggcagg acagcaaggg 3960ggaggattgg gaagacaata gcaggcatgc
tggggatgcg gtgggctcta tggcttctga 4020ggcggaaaga accagctagc
tcgatcgagg caggcagaag tatgcaaagc atgcatctca 4080attagtcagc
aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
4140gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc
atcccgcccc 4200taactccgcc cagttccgcc cattctccgc cccatggctg
actaattttt tttatttatg 4260cagaggccga ggccgcctct gcctctgagc
tattccagaa gtagtgagga ggcttttttg 4320gaggcctagg cttttgcaaa
aagctcccgg gagcttgtat atccattttc ggatctgatc 4380agcacgtgtt
gacaattaat catcggcata gtatatcggc atagtataat acgacaaggt
4440gaggaactaa accatggcca agttgaccag tgccgttccg gtgctcaccg
cgcgcgacgt 4500cgccggagcg gtcgagttct ggaccgaccg gctcgggttc
tcccgggact tcgtggagga 4560cgacttcgcc ggtgtggtcc gggacgacgt
gaccctgttc atcagcgcgg tccaggacca 4620ggtggtgccg gacaacaccc
tggcctgggt gtgggtgcgc ggcctggacg agctgtacgc 4680cgagtggtcg
gaggtcgtgt ccacgaactt ccgggacgcc tccgggccgg ccatgaccga
4740gatcggcgag cagccgtggg ggcgggagtt cgccctgcgc gacccggccg
gcaactgcgt 4800gcacttcgtg gccgaggagc aggactgaca cgtgctacga
gatttcgatt ccaccgccgc 4860cttctatgaa aggttgggct tcggaatcgt
tttccgggac gccggctgga tgatcctcca 4920gcgcggggat ctcatgctgg
agttcttcgc ccaccccaac ttgtttattg cagcttataa 4980tggttacaaa
taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca
5040ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgaa
ttttgcatta 5100atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt
gggcgctctt ccgcttcctc 5160gctcactgac tcgctgcgct cggtcgttcg
gctgcggcga gcggtatcag ctcactcaaa 5220ggcggtaata cggttatcca
cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa 5280aggccagcaa
aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
5340ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
gaaacccgac 5400aggactataa agataccagg cgtttccccc tggaagctcc
ctcgtgcgct ctcctgttcc 5460gaccctgccg cttaccggat acctgtccgc
ctttctccct tcgggaagcg tggcgctttc 5520tcaatgctca cgctgtaggt
atctcagttc ggtgtaggtc gttcgctcca agctgggctg 5580tgtgcacgaa
ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
5640gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
acaggattag 5700cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
tggtggccta actacggcta 5760cactagaagg acagtatttg gtatctgcgc
tctgctgaag ccagttacct tcggaaaaag 5820agttggtagc tcttgatccg
gcaaacaaac caccgctggt agcggtggtt tttttgtttg 5880caagcagcag
attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
5940ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
tgagattatc 6000aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
agttttaaat caatctaaag 6060tatatatgag taaacttggt ctgacagtta
ccaatgctta atcagtgagg cacctatctc 6120agcgatctgt ctatttcgtt
catccatagt tgcctgactc cccgtcgtgt agataactac 6180gatacgggag
ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
6240accggctcca gatttatcag caataaacca gccagccgga agggccgagc
gcagaagtgg 6300tcctgcaact ttatccgcct ccatccagtc tattaattgt
tgccgggaag ctagagtaag 6360tagttcgcca gttaatagtt tgcgcaacgt
tgttgccatt gctacaggca tcgtggtgtc 6420acgctcgtcg tttggtatgg
cttcattcag ctccggttcc caacgatcaa ggcgagttac 6480atgatccccc
atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
6540aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
attctcttac 6600tgtcatgcca tccgtaagat gcttttctgt gactggtgag
tactcaacca agtcattctg 6660agaatagtgt atgcggcgac cgagttgctc
ttgcccggcg tcaatacggg ataataccgc 6720gccacatagc agaactttaa
aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact 6780ctcaaggatc
ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
6840atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
gaaggcaaaa 6900tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
atactcatac tcttcctttt 6960tcaatattat tgaagcattt atcagggtta
ttgtctcatg agcggataca tatttgaatg 7020tatttagaaa aataaacaaa
taggggttcc gcgcacattt ccccgaaaag tgccacctga 7080cgtcga
7086378751DNAartificialnatleader-hsLINGO-1-Fc/pRS5a 37tcgacggatc
gggagatccg ggacatgtac ctcccagggg cccaggaaga ctacgggagg 60ctacaccaac
gtcaatcaga ggggcctgtg tagctaccga taagcggacc ctcaagaggg
120cattagcaat agtgtttata aggccccctt gttaacccta aacgggtagc
atatgcttcc 180cgggtagtag tatatactat ccagactaac cctaattcaa
tagcatatgt tacccaacgg 240gaagcatatg ctatcgaatt agggttagta
aaagggtcct aaggaacagc gatatctccc 300accccatgag ctgtcacggt
tttatttaca tggggtcagg attccacgag ggtagtgaac 360cattttagtc
acaagggcag tggctgaaga tcaaggagcg ggcagtgaac tctcctgaat
420cttcgcctgc ttcttcattc tccttcgttt agctaataga ataactgctg
agttgtgaac 480agtaaggtgt atgtgaggtg ctcgaaaaca aggtttcagg
tgacgccccc agaataaaat 540ttggacgggg ggttcagtgg tggcattgtg
ctatgacacc aatataaccc tcacaaaccc 600cttgggcaat aaatactagt
gtaggaatga aacattctga atatctttaa caatagaaat 660ccatggggtg
gggacaagcc gtaaagactg gatgtccatc tcacacgaat ttatggctat
720gggcaacaca taatcctagt gcaatatgat actggggtta ttaagatgtg
tcccaggcag 780ggaccaagac aggtgaacca tgttgttaca ctctatttgt
aacaagggga aagagagtgg 840acgccgacag cagcggactc cactggttgt
ctctaacacc cccgaaaatt aaacggggct 900ccacgccaat ggggcccata
aacaaagaca agtggccact cttttttttg aaattgtgga 960gtgggggcac
gcgtcagccc ccacacgccg ccctgcggtt ttggactgta aaataagggt
1020gtaataactt ggctgattgt aaccccgcta accactgcgg tcaaaccact
tgcccacaaa 1080accactaatg gcaccccggg gaatacctgc ataagtaggt
gggcgggcca agataggggc 1140gcgattgctg cgatctggag gacaaattac
acacacttgc gcctgagcgc caagcacagg 1200gttgttggtc ctcatattca
cgaggtcgct gagagcacgg tgggctaatg ttgccatggg 1260tagcatatac
tacccaaata tctggatagc atatgctatc ctaatctata tctgggtagc
1320ataggctatc ctaatctata tctgggtagc atatgctatc ctaatctata
tctgggtagt 1380atatgctatc ctaatttata tctgggtagc ataggctatc
ctaatctata tctgggtagc 1440atatgctatc ctaatctata tctgggtagt
atatgctatc ctaatctgta tccgggtagc 1500atatgctatc ctaatagaga
ttagggtagt atatgctatc ctaatttata tctgggtagc 1560atatactacc
caaatatctg gatagcatat gctatcctaa tctatatctg ggtagcatat
1620gctatcctaa tctatatctg ggtagcatag gctatcctaa tctatatctg
ggtagcatat 1680gctatcctaa tctatatctg ggtagtatat gctatcctaa
tttatatctg ggtagcatag 1740gctatcctaa tctatatctg ggtagcatat
gctatcctaa tctatatctg ggtagtatat 1800gctatcctaa tctgtatccg
ggtagcatat gctatcctca tgcatataca gtcagcatat 1860gatacccagt
agtagagtgg gagtgctatc ctttgcatat gccgccacct cccaaggggg
1920cgtgaatttt cgctgcttgt ccttttcctg catgcggatc ttcaatattg
gccattagcc 1980atattattca
ttggttatat agcataaatc aatattggct attggccatt gcatacgttg
2040tatctatatc ataatatgta catttatatt ggctcatgtc caatatgacc
gccatgttgg 2100cattgattat tgactagtta ttaatagtaa tcaattacgg
ggtcattagt tcatagccca 2160tatatggagt tccgcgttac ataacttacg
gtaaatggcc cgcctggctg accgcccaac 2220gacccccgcc cattgacgtc
aataatgacg tatgttccca tagtaacgcc aatagggact 2280ttccattgac
gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa
2340gtgtatcata tgccaagtcc gccccctatt gacgtcaatg acggtaaatg
gcccgcctgg 2400cattatgccc agtacatgac cttacgggac tttcctactt
ggcagtacat ctacgtatta 2460gtcatcgcta ttaccatggt gatgcggttt
tggcagtaca ccaatgggcg tggatagcgg 2520tttgactcac ggggatttcc
aagtctccac cccattgacg tcaatgggag tttgttttgg 2580caccaaaatc
aacgggactt tccaaaatgt cgtaataacc ccgccccgtt gacgcaaatg
2640ggcggtaggc gtgtacggtg ggaggtctat ataagcagag ctcgtttagt
gaaccgtcag 2700atcgcctgga gacgccatcc acgctgtttt gacctccata
gaagacaccg ggaccgatcc 2760agcctccgcg gccgggaacg gtgcattgga
acgcggattc cccgtgccaa gagtgacgta 2820agtaccgcct atagagtcta
taggcccacc cccttggctt cgttagaacg cggctacaat 2880taatacataa
ccttatgtat catacacata cgatttaggt gacactatag aataacatcc
2940actttgcctt tctctccaca ggtgtccact cccaggtcca actgcacgga
agcttgccgc 3000caccatgcag gtgagcaaga ggatgctggc ggggggcgtg
aggagcatgc ccagccccct 3060cctggcctgc tggcagccca tcctcctgct
ggtgctgggc tcagtgctgt caggctcggc 3120cacgggctgc ccgccccgct
gcgagtgctc cgcccaggac cgcgctgtgc tgtgccaccg 3180caagcgcttt
gtggcagtcc ccgagggcat ccccaccgag acgcgcctgc tggacctagg
3240caagaaccgc atcaaaacgc tcaaccagga cgagttcgcc agcttcccgc
acctggagga 3300gctggagctc aacgagaaca tcgtgagcgc cgtggagccc
ggcgccttca acaacctctt 3360caacctccgg acgctgggtc tccgcagcaa
ccgcctgaag ctcatcccgc taggcgtctt 3420cactggcctc agcaacctga
ccaagctgga catcagcgag aacaagatcg ttatcctact 3480ggactacatg
tttcaggacc tgtacaacct caagtcactg gaggttggcg acaatgacct
3540cgtctacatc tctcaccgcg ccttcagcgg cctcaacagc ctggagcagc
tgacgctgga 3600gaaatgcaac ctgacctcca tccccaccga ggcgctgtcc
cacctgcacg gcctcatcgt 3660cctgaggctc cggcacctca acatcaatgc
catccgggac tactccttca agaggctgta 3720ccgactcaag gtcttggaga
tctcccactg gccctacttg gacaccatga cacccaactg 3780cctctacggc
ctcaacctga cgtccctgtc catcacacac tgcaatctga ccgctgtgcc
3840ctacctggcc gtccgccacc tagtctatct ccgcttcctc aacctctcct
acaaccccat 3900cagcaccatt gagggctcca tgttgcatga gctgctccgg
ctgcaggaga tccagctggt 3960gggcgggcag ctggccgtgg tggagcccta
tgccttccgc ggcctcaact acctgcgcgt 4020gctcaatgtc tctggcaacc
agctgaccac actggaggaa tcagtcttcc actcggtggg 4080caacctggag
acactcatcc tggactccaa cccgctggcc tgcgactgtc ggctcctgtg
4140ggtgttccgg cgccgctggc ggctcaactt caaccggcag cagcccacgt
gcgccacgcc 4200cgagtttgtc cagggcaagg agttcaagga cttccctgat
gtgctactgc ccaactactt 4260cacctgccgc cgcgcccgca tccgggaccg
caaggcccag caggtgtttg tggacgaggg 4320ccacacggtg cagtttgtgt
gccgggccga tggcgacccg ccgcccgcca tcctctggct 4380ctcaccccga
aagcacctgg tctcagccaa gagcaatggg cggctcacag tcttccctga
4440tggcacgctg gaggtgcgct acgcccaggt acaggacaac ggcacgtacc
tgtgcatcgc 4500ggccaacgcg ggcggcaacg actccatgcc cgcccacctg
catgtgcgca gctactcgcc 4560cgactggccc catcagccca acaagacctt
cgctttcatc tccaaccagc cgggcgaggg 4620agaggccaac agcacccgcg
ccactgtgcc tttccccttc gacatcaaga ccctcgaggt 4680tctgttccag
ggtccgaaat cttgtgacaa aactcacaca tgcccaccgt gcccagcacc
4740tgaactcctg gggggaccgt cagtcttcct cttcccccca aaacccaagg
acaccctcat 4800gatctcccgg acccctgagg tcacatgcgt ggtggtggac
gtgagccacg aagaccctga 4860ggtcaagttc aactggtacg tggacggcgt
ggaggtgcat aatgccaaga caaagccgcg 4920ggaggagcag tacaacagca
cgtaccgtgt ggtcagcgtc ctcaccgtcc tgcaccagga 4980ctggctgaat
ggcaaggagt acaagtgcaa ggtctccaac aaagccctcc cagcccccat
5040cgagaaaacc atctccaaag ccaaagggca gccccgagaa ccacaggtgt
acaccctgcc 5100cccatcccgg gatgagctga ccaagaacca ggtcagcctg
acctgcctgg tcaaaggctt 5160ctatcccagc gacatcgccg tggagtggga
gagcaatggg cagccggaga acaactacaa 5220gaccacgcct cccgtgctgg
actccgacgg ctccttcttc ctctacagca agctcaccgt 5280ggacaagagc
aggtggcagc aggggaacgt cttctcatgc tccgtgatgc atgaggctct
5340gcacaaccac tacacgcaga agagcctctc cctgtctccg ggtaaatgag
atctggtacc 5400tcgcgatggc ggccgctcta gagggcccgt ttaaacccgc
tgatcagcct cgactgtgcc 5460ttctagttgc cagccatctg ttgtttgccc
ctcccccgtg ccttccttga ccctggaagg 5520tgccactccc actgtccttt
cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag 5580gtgtcattct
attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga
5640caatagcagg catgctgggg atgcggtggg ctctatggct tctgaggcgg
aaagaaccag 5700ctagctcgat cgaggcaggc agaagtatgc aaagcatgca
tctcaattag tcagcaacca 5760ggtgtggaaa gtccccaggc tccccagcag
gcagaagtat gcaaagcatg catctcaatt 5820agtcagcaac catagtcccg
cccctaactc cgcccatccc gcccctaact ccgcccagtt 5880ccgcccattc
tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg
5940cctctgcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc
ctaggctttt 6000gcaaaaagct cccgggagct tgtatatcca ttttcggatc
tgatcagcac gtgttgacaa 6060ttaatcatcg gcatagtata tcggcatagt
ataatacgac aaggtgagga actaaaccat 6120ggccaagttg accagtgccg
ttccggtgct caccgcgcgc gacgtcgccg gagcggtcga 6180gttctggacc
gaccggctcg ggttctcccg ggacttcgtg gaggacgact tcgccggtgt
6240ggtccgggac gacgtgaccc tgttcatcag cgcggtccag gaccaggtgg
tgccggacaa 6300caccctggcc tgggtgtggg tgcgcggcct ggacgagctg
tacgccgagt ggtcggaggt 6360cgtgtccacg aacttccggg acgcctccgg
gccggccatg accgagatcg gcgagcagcc 6420gtgggggcgg gagttcgccc
tgcgcgaccc ggccggcaac tgcgtgcact tcgtggccga 6480ggagcaggac
tgacacgtgc tacgagattt cgattccacc gccgccttct atgaaaggtt
6540gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg
gggatctcat 6600gctggagttc ttcgcccacc ccaacttgtt tattgcagct
tataatggtt acaaataaag 6660caatagcatc acaaatttca caaataaagc
atttttttca ctgcattcta gttgtggttt 6720gtccaaactc atcaatgtat
cttatcatgt ctgaattttg cattaatgaa tcggccaacg 6780cgcggggaga
ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
6840gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
taatacggtt 6900atccacagaa tcaggggata acgcaggaaa gaacatgtga
gcaaaaggcc agcaaaaggc 6960caggaaccgt aaaaaggccg cgttgctggc
gtttttccat aggctccgcc cccctgacga 7020gcatcacaaa aatcgacgct
caagtcagag gtggcgaaac ccgacaggac tataaagata 7080ccaggcgttt
ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
7140cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcaat
gctcacgctg 7200taggtatctc agttcggtgt aggtcgttcg ctccaagctg
ggctgtgtgc acgaaccccc 7260cgttcagccc gaccgctgcg ccttatccgg
taactatcgt cttgagtcca acccggtaag 7320acacgactta tcgccactgg
cagcagccac tggtaacagg attagcagag cgaggtatgt 7380aggcggtgct
acagagttct tgaagtggtg gcctaactac ggctacacta gaaggacagt
7440atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
gtagctcttg 7500atccggcaaa caaaccaccg ctggtagcgg tggttttttt
gtttgcaagc agcagattac 7560gcgcagaaaa aaaggatctc aagaagatcc
tttgatcttt tctacggggt ctgacgctca 7620gtggaacgaa aactcacgtt
aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac 7680ctagatcctt
ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
7740ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
tctgtctatt 7800tcgttcatcc atagttgcct gactccccgt cgtgtagata
actacgatac gggagggctt 7860accatctggc cccagtgctg caatgatacc
gcgagaccca cgctcaccgg ctccagattt 7920atcagcaata aaccagccag
ccggaagggc cgagcgcaga agtggtcctg caactttatc 7980cgcctccatc
cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
8040tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
cgtcgtttgg 8100tatggcttca ttcagctccg gttcccaacg atcaaggcga
gttacatgat cccccatgtt 8160gtgcaaaaaa gcggttagct ccttcggtcc
tccgatcgtt gtcagaagta agttggccgc 8220agtgttatca ctcatggtta
tggcagcact gcataattct cttactgtca tgccatccgt 8280aagatgcttt
tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
8340gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
atagcagaac 8400tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
aaactctcaa ggatcttacc 8460gctgttgaga tccagttcga tgtaacccac
tcgtgcaccc aactgatctt cagcatcttt 8520tactttcacc agcgtttctg
ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg 8580aataagggcg
acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
8640catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
agaaaaataa 8700acaaataggg gttccgcgca catttccccg aaaagtgcca
cctgacgtcg a
8751387830DNAartificialIgleader-hsLINGO-1-?LRR-Fc/pRS5a
38tcgacggatc gggagatccg ggacatgtac ctcccagggg cccaggaaga ctacgggagg
60ctacaccaac gtcaatcaga ggggcctgtg tagctaccga taagcggacc ctcaagaggg
120cattagcaat agtgtttata aggccccctt gttaacccta aacgggtagc
atatgcttcc 180cgggtagtag tatatactat ccagactaac cctaattcaa
tagcatatgt tacccaacgg 240gaagcatatg ctatcgaatt agggttagta
aaagggtcct aaggaacagc gatatctccc 300accccatgag ctgtcacggt
tttatttaca tggggtcagg attccacgag ggtagtgaac 360cattttagtc
acaagggcag tggctgaaga tcaaggagcg ggcagtgaac tctcctgaat
420cttcgcctgc ttcttcattc tccttcgttt agctaataga ataactgctg
agttgtgaac 480agtaaggtgt atgtgaggtg ctcgaaaaca aggtttcagg
tgacgccccc agaataaaat 540ttggacgggg ggttcagtgg tggcattgtg
ctatgacacc aatataaccc tcacaaaccc 600cttgggcaat aaatactagt
gtaggaatga aacattctga atatctttaa caatagaaat 660ccatggggtg
gggacaagcc gtaaagactg gatgtccatc tcacacgaat ttatggctat
720gggcaacaca taatcctagt gcaatatgat actggggtta ttaagatgtg
tcccaggcag 780ggaccaagac aggtgaacca tgttgttaca ctctatttgt
aacaagggga aagagagtgg 840acgccgacag cagcggactc cactggttgt
ctctaacacc cccgaaaatt aaacggggct 900ccacgccaat ggggcccata
aacaaagaca agtggccact cttttttttg aaattgtgga 960gtgggggcac
gcgtcagccc ccacacgccg ccctgcggtt ttggactgta aaataagggt
1020gtaataactt ggctgattgt aaccccgcta accactgcgg tcaaaccact
tgcccacaaa 1080accactaatg gcaccccggg gaatacctgc ataagtaggt
gggcgggcca agataggggc 1140gcgattgctg cgatctggag gacaaattac
acacacttgc gcctgagcgc caagcacagg 1200gttgttggtc ctcatattca
cgaggtcgct gagagcacgg tgggctaatg ttgccatggg 1260tagcatatac
tacccaaata tctggatagc atatgctatc ctaatctata tctgggtagc
1320ataggctatc ctaatctata tctgggtagc atatgctatc ctaatctata
tctgggtagt 1380atatgctatc ctaatttata tctgggtagc ataggctatc
ctaatctata tctgggtagc 1440atatgctatc ctaatctata tctgggtagt
atatgctatc ctaatctgta tccgggtagc 1500atatgctatc ctaatagaga
ttagggtagt atatgctatc ctaatttata tctgggtagc 1560atatactacc
caaatatctg gatagcatat gctatcctaa tctatatctg ggtagcatat
1620gctatcctaa tctatatctg ggtagcatag gctatcctaa tctatatctg
ggtagcatat 1680gctatcctaa tctatatctg ggtagtatat gctatcctaa
tttatatctg ggtagcatag 1740gctatcctaa tctatatctg ggtagcatat
gctatcctaa tctatatctg ggtagtatat 1800gctatcctaa tctgtatccg
ggtagcatat gctatcctca tgcatataca gtcagcatat 1860gatacccagt
agtagagtgg gagtgctatc ctttgcatat gccgccacct cccaaggggg
1920cgtgaatttt cgctgcttgt ccttttcctg catgcggatc ttcaatattg
gccattagcc 1980atattattca ttggttatat agcataaatc aatattggct
attggccatt gcatacgttg 2040tatctatatc ataatatgta catttatatt
ggctcatgtc caatatgacc gccatgttgg 2100cattgattat tgactagtta
ttaatagtaa tcaattacgg ggtcattagt tcatagccca 2160tatatggagt
tccgcgttac ataacttacg gtaaatggcc cgcctggctg accgcccaac
2220gacccccgcc cattgacgtc aataatgacg tatgttccca tagtaacgcc
aatagggact 2280ttccattgac gtcaatgggt ggagtattta cggtaaactg
cccacttggc agtacatcaa 2340gtgtatcata tgccaagtcc gccccctatt
gacgtcaatg acggtaaatg gcccgcctgg 2400cattatgccc agtacatgac
cttacgggac tttcctactt ggcagtacat ctacgtatta 2460gtcatcgcta
ttaccatggt gatgcggttt tggcagtaca ccaatgggcg tggatagcgg
2520tttgactcac ggggatttcc aagtctccac cccattgacg tcaatgggag
tttgttttgg 2580caccaaaatc aacgggactt tccaaaatgt cgtaataacc
ccgccccgtt gacgcaaatg 2640ggcggtaggc gtgtacggtg ggaggtctat
ataagcagag ctcgtttagt gaaccgtcag 2700atcgcctgga gacgccatcc
acgctgtttt gacctccata gaagacaccg ggaccgatcc 2760agcctccgcg
gccgggaacg gtgcattgga acgcggattc cccgtgccaa gagtgacgta
2820agtaccgcct atagagtcta taggcccacc cccttggctt cgttagaacg
cggctacaat 2880taatacataa ccttatgtat catacacata cgatttaggt
gacactatag aataacatcc 2940actttgcctt tctctccaca ggtgtccact
cccaggtcca actgcacgga agcttgccgc 3000caccatgagt gtgctcactc
aggtcctggc gttgctgctg ctgtggctta caggtacgcg 3060ttgtacgggc
tgcccgcccc gctgcgagtg ctccgcccag gaccgcgctg tgctgtgcca
3120ccgcaagcgc tttgtggcag tccccgaggg catccccacc aacctggaga
cactcatcct 3180ggactccaac ccgctggcct gcgactgtcg gctcctgtgg
gtgttccggc gccgctggcg 3240gctcaacttc aaccggcagc agcccacgtg
cgccacgccc gagtttgtcc agggcaagga 3300gttcaaggac ttccctgatg
tgctactgcc caactacttc acctgccgcc gcgcccgcat 3360ccgggaccgc
aaggcccagc aggtgtttgt ggacgagggc cacacggtgc agtttgtgtg
3420ccgggccgat ggcgacccgc cgcccgccat cctctggctc tcaccccgaa
agcacctggt 3480ctcagccaag agcaatgggc ggctcacagt cttccctgat
ggcacgctgg aggtgcgcta 3540cgcccaggta caggacaacg gcacgtacct
gtgcatcgcg gccaacgcgg gcggcaacga 3600ctccatgccc gcccacctgc
atgtgcgcag ctactcgccc gactggcccc atcagcccaa 3660caagaccttc
gctttcatct ccaaccagcc gggcgaggga gaggccaaca gcacccgcgc
3720cactgtgcct ttccccttcg acatcaagac cctcgaggtt ctgttccagg
gtccgaaatc 3780ttgtgacaaa actcacacat gcccaccgtg cccagcacct
gaactcctgg ggggaccgtc 3840agtcttcctc ttccccccaa aacccaagga
caccctcatg atctcccgga cccctgaggt 3900cacatgcgtg gtggtggacg
tgagccacga agaccctgag gtcaagttca actggtacgt 3960ggacggcgtg
gaggtgcata atgccaagac aaagccgcgg gaggagcagt acaacagcac
4020gtaccgtgtg gtcagcgtcc tcaccgtcct gcaccaggac tggctgaatg
gcaaggagta 4080caagtgcaag gtctccaaca aagccctccc agcccccatc
gagaaaacca tctccaaagc 4140caaagggcag ccccgagaac cacaggtgta
caccctgccc ccatcccggg atgagctgac 4200caagaaccag gtcagcctga
cctgcctggt caaaggcttc tatcccagcg acatcgccgt 4260ggagtgggag
agcaatgggc agccggagaa caactacaag accacgcctc ccgtgctgga
4320ctccgacggc tccttcttcc tctacagcaa gctcaccgtg gacaagagca
ggtggcagca 4380ggggaacgtc ttctcatgct ccgtgatgca tgaggctctg
cacaaccact acacgcagaa 4440gagcctctcc ctgtctccgg gtaaatgaga
tctggtacct cgcgatggcg gccgctctag 4500agggcccgtt taaacccgct
gatcagcctc gactgtgcct tctagttgcc agccatctgt 4560tgtttgcccc
tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc
4620ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta
ttctgggggg 4680tggggtgggg caggacagca agggggagga ttgggaagac
aatagcaggc atgctgggga 4740tgcggtgggc tctatggctt ctgaggcgga
aagaaccagc tagctcgatc gaggcaggca 4800gaagtatgca aagcatgcat
ctcaattagt cagcaaccag gtgtggaaag tccccaggct 4860ccccagcagg
cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc
4920ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct
ccgccccatg 4980gctgactaat tttttttatt tatgcagagg ccgaggccgc
ctctgcctct gagctattcc 5040agaagtagtg aggaggcttt tttggaggcc
taggcttttg caaaaagctc ccgggagctt 5100gtatatccat tttcggatct
gatcagcacg tgttgacaat taatcatcgg catagtatat 5160cggcatagta
taatacgaca aggtgaggaa ctaaaccatg gccaagttga ccagtgccgt
5220tccggtgctc accgcgcgcg acgtcgccgg agcggtcgag ttctggaccg
accggctcgg 5280gttctcccgg gacttcgtgg aggacgactt cgccggtgtg
gtccgggacg acgtgaccct 5340gttcatcagc gcggtccagg accaggtggt
gccggacaac accctggcct gggtgtgggt 5400gcgcggcctg gacgagctgt
acgccgagtg gtcggaggtc gtgtccacga acttccggga 5460cgcctccggg
ccggccatga ccgagatcgg cgagcagccg tgggggcggg agttcgccct
5520gcgcgacccg gccggcaact gcgtgcactt cgtggccgag gagcaggact
gacacgtgct 5580acgagatttc gattccaccg ccgccttcta tgaaaggttg
ggcttcggaa tcgttttccg 5640ggacgccggc tggatgatcc tccagcgcgg
ggatctcatg ctggagttct tcgcccaccc 5700caacttgttt attgcagctt
ataatggtta caaataaagc aatagcatca caaatttcac 5760aaataaagca
tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc
5820ttatcatgtc tgaattttgc attaatgaat cggccaacgc gcggggagag
gcggtttgcg 5880tattgggcgc tcttccgctt cctcgctcac tgactcgctg
cgctcggtcg ttcggctgcg 5940gcgagcggta tcagctcact caaaggcggt
aatacggtta tccacagaat caggggataa 6000cgcaggaaag aacatgtgag
caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc 6060gttgctggcg
tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
6120aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
cccctggaag 6180ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
ggatacctgt ccgcctttct 6240cccttcggga agcgtggcgc tttctcaatg
ctcacgctgt aggtatctca gttcggtgta 6300ggtcgttcgc tccaagctgg
gctgtgtgca cgaacccccc gttcagcccg accgctgcgc 6360cttatccggt
aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
6420agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
cagagttctt 6480gaagtggtgg cctaactacg gctacactag aaggacagta
tttggtatct gcgctctgct 6540gaagccagtt accttcggaa aaagagttgg
tagctcttga tccggcaaac aaaccaccgc 6600tggtagcggt ggtttttttg
tttgcaagca gcagattacg cgcagaaaaa aaggatctca 6660agaagatcct
ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
6720agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
taaattaaaa 6780atgaagtttt aaatcaatct aaagtatata tgagtaaact
tggtctgaca gttaccaatg 6840cttaatcagt gaggcaccta tctcagcgat
ctgtctattt cgttcatcca tagttgcctg 6900actccccgtc gtgtagataa
ctacgatacg ggagggctta ccatctggcc ccagtgctgc 6960aatgataccg
cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
7020cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
agtctattaa 7080ttgttgccgg gaagctagag taagtagttc gccagttaat
agtttgcgca acgttgttgc 7140cattgctaca ggcatcgtgg tgtcacgctc
gtcgtttggt atggcttcat tcagctccgg 7200ttcccaacga tcaaggcgag
ttacatgatc ccccatgttg tgcaaaaaag cggttagctc 7260cttcggtcct
ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
7320ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
ctgtgactgg 7380tgagtactca accaagtcat tctgagaata gtgtatgcgg
cgaccgagtt gctcttgccc 7440ggcgtcaata cgggataata ccgcgccaca
tagcagaact ttaaaagtgc tcatcattgg 7500aaaacgttct tcggggcgaa
aactctcaag gatcttaccg ctgttgagat ccagttcgat 7560gtaacccact
cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
7620gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
cacggaaatg 7680ttgaatactc atactcttcc tttttcaata ttattgaagc
atttatcagg gttattgtct 7740catgagcgga tacatatttg aatgtattta
gaaaaataaa caaatagggg ttccgcgcac 7800atttccccga aaagtgccac
ctgacgtcga 7830395025DNAartificialFab Expression Vector
pMORPH-X9_MH 39ctagataacg agggcaaaaa atgaaaaaga cagctatcgc
gattgcagtg gcactggctg 60gtttcgctac cgtagcgcag gccgatatcg tgctgaccca
gccgccttca gtgagtggcg 120caccaggtca gcgtgtgacc atctcgtgta
gcggcagcag cagcaacatt ggtaataatt 180atgtgtcttg gtaccagcag
ttgcccggga cggcgccgaa acttctgatt tctcgtaatt 240ctaagcgtcc
ctcaggcgtg ccggatcgtt ttagcggatc caaaagcggc accagcgcga
300gccttgcgat
tacgggcctg caaagcgaag acgaagcgga ttattattgc tctacttatg
360atactttttc tattgtgttt ggcggcggca cgaagttaac cgttcttggc
cagccgaaag 420ccgcaccgag tgtgacgctg tttccgccga gcagcgaaga
attgcaggcg aacaaagcga 480ccctggtgtg cctgattagc gacttttatc
cgggagccgt gacagtggcc tggaaggcag 540atagcagccc cgtcaaggcg
ggagtggaga ccaccacacc ctccaaacaa agcaacaaca 600agtacgcggc
cagcagctat ctgagcctga cgcctgagca gtggaagtcc cacagaagct
660acagctgcca ggtcacgcat gaggggagca ccgtggaaaa aaccgttgcg
ccgactgagg 720cctgataagc atgcgtagga gaaaataaaa tgaaacaaag
cactattgca ctggcactct 780taccgttgct cttcacccct gttaccaaag
cccaggtgca attgcaacag tctggtccgg 840gcctggtgaa accgagccaa
accctgagcc tgacctgtgc gatttccgga gatagcgtga 900gcgataattc
tgctgcttgg tcttggattc gccagtctcc tgggcgtggc ctcgagtggc
960tgggccttat ctatcttcgt agcaagtggg ataacgatta tgcggtgagc
gtgaaaagcc 1020ggattaccat caacccggat acttcgaaaa accagtttag
cctgcaactg aacagcgtga 1080ccccggaaga tacggccgtg tattattgcg
cgcgtactgg tcgtgctgat gagtttgatg 1140tttggggcca aggcaccctg
gtgacggtta gctcagcgtc gaccaaaggt ccaagcgtgt 1200ttccgctggc
tccgagcagc aaaagcacca gcggcggcac ggctgccctg ggctgcctgg
1260ttaaagatta tttcccggaa ccagtcaccg tgagctggaa cagcggggcg
ctgaccagcg 1320gcgtgcatac ctttccggcg gtgctgcaaa gcagcggcct
gtatagcctg agcagcgttg 1380tgaccgtgcc gagcagcagc ttaggcactc
agacctatat ttgcaacgtg aaccataaac 1440cgagcaacac caaagtggat
aaaaaagtgg aaccgaaaag cgaattcgag cagaagctga 1500tctctgagga
ggatctgaac ggcgcgccgc accatcatca ccatcactga taagcttgac
1560ctgtgaagtg aaaaatggcg cagattgtgc gacatttttt ttgtctgccg
tttaattaaa 1620gggggggggg ggccggcctg ggggggggtg tacatgaaat
tgtaaacgtt aatattttgt 1680taaaattcgc gttaaatttt tgttaaatca
gctcattttt taaccaatag gccgaaatcg 1740gcaaaatccc ttataaatca
aaagaataga ccgagatagg gttgagtgtt gttccagttt 1800ggaacaagag
tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct
1860atcagggcga tggcccacta cgagaaccat caccctaatc aagttttttg
gggtcgaggt 1920gccgtaaagc actaaatcgg aaccctaaag ggagcccccg
atttagagct tgacggggaa 1980agccggcgaa cgtggcgaga aaggaaggga
agaaagcgaa aggagcgggc gctagggcgc 2040tggcaagtgt agcggtcacg
ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc 2100tacagggcgc
gtgctagact agtgtttaaa ccggaccggg ggggggctta agtgggctgc
2160aaaacaaaac ggcctcctgt caggaagccg cttttatcgg gtagcctcac
tgcccgcttt 2220ccagtcggga aacctgtcgt gccagctgca tcagtgaatc
ggccaacgcg cggggagagg 2280cggtttgcgt attgggagcc agggtggttt
ttcttttcac cagtgagacg ggcaacagct 2340gattgccctt caccgcctgg
ccctgagaga gttgcagcaa gcggtccacg ctggtttgcc 2400ccagcaggcg
aaaatcctgt ttgatggtgg tcagcggcgg gatataacat gagctgtcct
2460cggtatcgtc gtatcccact accgagatgt ccgcaccaac gcgcagcccg
gactcggtaa 2520tggcacgcat tgcgcccagc gccatctgat cgttggcaac
cagcatcgca gtgggaacga 2580tgccctcatt cagcatttgc atggtttgtt
gaaaaccgga catggcactc cagtcgcctt 2640cccgttccgc tatcggctga
atttgattgc gagtgagata tttatgccag ccagccagac 2700gcagacgcgc
cgagacagaa cttaatgggc cagctaacag cgcgatttgc tggtggccca
2760atgcgaccag atgctccacg cccagtcgcg taccgtcctc atgggagaaa
ataatactgt 2820tgatgggtgt ctggtcagag acatcaagaa ataacgccgg
aacattagtg caggcagctt 2880ccacagcaat agcatcctgg tcatccagcg
gatagttaat aatcagccca ctgacacgtt 2940gcgcgagaag attgtgcacc
gccgctttac aggcttcgac gccgcttcgt tctaccatcg 3000acacgaccac
gctggcaccc agttgatcgg cgcgagattt aatcgccgcg acaatttgcg
3060acggcgcgtg cagggccaga ctggaggtgg caacgccaat cagcaacgac
tgtttgcccg 3120ccagttgttg tgccacgcgg ttaggaatgt aattcagctc
cgccatcgcc gcttccactt 3180tttcccgcgt tttcgcagaa acgtggctgg
cctggttcac cacgcgggaa acggtctgat 3240aagagacacc ggcatactct
gcgacatcgt ataacgttac tggtttcaca ttcaccaccc 3300tgaattgact
ctcttccggg cgctatcatg ccataccgcg aaaggttttg cgccattcga
3360tgctagccat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
gccgcgttgc 3420tggcgttttt ccataggctc cgcccccctg acgagcatca
caaaaatcga cgctcaagtc 3480agaggtggcg aaacccgaca ggactataaa
gataccaggc gtttccccct ggaagctccc 3540tcgtgcgctc tcctgttccg
accctgccgc ttaccggata cctgtccgcc tttctccctt 3600cgggaagcgt
ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
3660ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
tgcgccttat 3720ccggtaacta tcgtcttgag tccaacccgg taagacacga
cttatcgcca ctggcagcag 3780ccactggtaa caggattagc agagcgaggt
atgtaggcgg tgctacagag ttcttgaagt 3840ggtggcctaa ctacggctac
actagaagaa cagtatttgg tatctgcgct ctgctgtagc 3900cagttacctt
cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
3960gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
tctcaagaag 4020atcctttgat cttttctacg gggtctgacg ctcagtggaa
cgaaaactca cgttaaggga 4080ttttggtcag atctagcacc aggcgtttaa
gggcaccaat aactgcctta aaaaaattac 4140gccccgccct gccactcatc
gcagtactgt tgtaattcat taagcattct gccgacatgg 4200aagccatcac
aaacggcatg atgaacctga atcgccagcg gcatcagcac cttgtcgcct
4260tgcgtataat atttgcccat agtgaaaacg ggggcgaaga agttgtccat
attggctacg 4320tttaaatcaa aactggtgaa actcacccag ggattggctg
agacgaaaaa catattctca 4380ataaaccctt tagggaaata ggccaggttt
tcaccgtaac acgccacatc ttgcgaatat 4440atgtgtagaa actgccggaa
atcgtcgtgg tattcactcc agagcgatga aaacgtttca 4500gtttgctcat
ggaaaacggt gtaacaaggg tgaacactat cccatatcac cagctcaccg
4560tctttcattg ccatacggaa ctccgggtga gcattcatca ggcgggcaag
aatgtgaata 4620aaggccggat aaaacttgtg cttatttttc tttacggtct
ttaaaaaggc cgtaatatcc 4680agctgaacgg tctggttata ggtacattga
gcaactgact gaaatgcctc aaaatgttct 4740ttacgatgcc attgggatat
atcaacggtg gtatatccag tgattttttt ctccatttta 4800gcttccttag
ctcctgaaaa tctcgataac tcaaaaaata cgcccggtag tgatcttatt
4860tcattatggt gaaagttgga acctcacccg acgtctaatg tgagttagct
cactcattag 4920gcaccccagg ctttacactt tatgcttccg gctcgtatgt
tgtgtggaat tgtgagcgga 4980taacaatttc acacaggaaa cagctatgac
catgattacg aattt 5025406746DNAartificialIgG4 Expression Vector
pMORPH?_h_Ig 4 40aattgcatga agaatctgct tagggttagg cgttttgcgc
tgcttcgcga tgtacgggcc 60agatatacgc gttgacattg attattgact agttattaat
agtaatcaat tacggggtca 120ttagttcata gcccatatat ggagttccgc
gttacataac ttacggtaaa tggcccgcct 180ggctgaccgc ccaacgaccc
ccgcccattg acgtcaataa tgacgtatgt tcccatagta 240acgccaatag
ggactttcca ttgacgtcaa tgggtggact atttacggta aactgcccac
300ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt
caatgacggt 360aaatggcccg cctggcatta tgcccagtac atgaccttat
gggactttcc tacttggcag 420tacatctacg tattagtcat cgctattacc
atggtgatgc ggttttggca gtacatcaat 480gggcgtggat agcggtttga
ctcacgggga tttccaagtc tccaccccat tgacgtcaat 540gggagtttgt
tttggcacca aaatcaacgg gactttccaa aatgtcgtaa caactccgcc
600ccattgacgc aaatgggcgg taggcgtgta cggtgggagg tctatataag
cagagctctc 660tggctaacta gagaacccac tgcttactgg cttatcgaaa
ttaatacgac tcactatagg 720gagacccaag ctggctagcg ccaccatgaa
acacctgtgg ttcttcctcc tgctggtggc 780agctcccaga tgggtcctgt
cccaggtgga attgcaacag tctggtccgg gcctggtgaa 840accgagccaa
accctgagcc tgacctgtgc gatttccgga gatagcgtga gcgataattc
900tgctgcttgg tcttggattc gccagtctcc tgggcgtggc ctcgagtggc
tgggccttat 960ctatcttcgt agcaagtggg ataacgatta tgcggtgagc
gtgaaaagcc ggattaccat 1020caacccggat acttcgaaaa accagtttag
cctgcaactg aacagcgtga ccccggaaga 1080tacggccgtg tattattgcg
cgcgtactgg tcgtgctgat gagtttgatg tttggggcca 1140aggcaccctg
gtgacggtta gctcagcttc caccaaggga ccatccgtct tccccctggc
1200gccctgctcc aggagcacct ccgagagcac agccgccctg ggctgcctgg
tcaaggacta 1260cttccccgaa ccggtgacgg tgtcgtggaa ctcaggcgcc
ctgaccagcg gcgtgcacac 1320cttcccggct gtcctacagt cctcaggact
ctactccctc agcagcgtgg tgaccgtgcc 1380ctccagcagc ttgggcacga
agacctacac ctgcaacgta gatcacaagc ccagcaacac 1440caaggtggac
aagagagttg agtccaaata tggtccccca tgcccatcat gcccagcacc
1500tgagttcctg gggggaccat cagtcttcct gttcccccca aaacccaagg
acactctcat 1560gatctcccgg acccctgagg tcacgtgcgt ggtggtggac
gtgagccagg aagaccccga 1620ggtccagttc aactggtacg tggatggcgt
ggaggtgcat aatgccaaga caaagccgcg 1680ggaggagcag ttcaacagca
cgtaccgtgt ggtcagcgtc ctcaccgtcc tgcaccagga 1740ctggctgaac
ggcaaggagt acaagtgcaa ggtctccaac aaaggcctcc cgtcctccat
1800cgagaaaacc atctccaaag ccaaagggca gccccgagag ccacaggtgt
acaccctgcc 1860cccatcccag gaggagatga ccaagaacca ggtcagcctg
acctgcctgg tcaaaggctt 1920ctaccccagc gacatcgccg tggagtggga
gagcaatggg cagccggaga acaactacaa 1980gaccacgcct cccgtgctgg
actccgacgg ctccttcttc ctctacagca ggctaaccgt 2040ggacaagagc
aggtggcagg aggggaatgt cttctcatgc tccgtgatgc atgaggctct
2100gcacaaccac tacacacaga agagcctctc cctgtctctg ggtaaatgag
ggcccgttta 2160aacccgctga tcagcctcga ctgtgccttc tagttgccag
ccatctgttg tttgcccctc 2220ccccgtgcct tccttgaccc tggaaggtgc
cactcccact gtcctttcct aataaaatga 2280ggaaattgca tcgcattgtc
tgagtaggtg tcattctatt ctggggggtg gggtggggca 2340ggacagcaag
ggggaggatt gggaagacaa tagcaggcat gctggggatg cggtgggctc
2400tatggcttct gaggcggaaa gaaccagctg gggctctagg gggtatcccc
acgcgccctg 2460tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc
agcgtgaccg ctacacttgc 2520cagcgcccta gcgcccgctc ctttcgcttt
cttcccttcc tttctcgcca cgttcgccgg 2580ctttccccgt caagctctaa
atcggggcat ccctttaggg ttccgattta gtgctttacg 2640gcacctcgac
cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg
2700atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg
gactcttgtt 2760ccaaactgga acaacactca accctatctc ggtctattct
tttgatttat aagggatttt 2820ggggatttcg gcctattggt taaaaaatga
gctgatttaa caaaaattta acgcgaatta 2880attctgtgga atgtgtgtca
gttagggtgt ggaaagtccc caggctcccc aggcaggcag 2940aagtatgcaa
agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc
3000cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
tagtcccgcc 3060cctaactccg cccatcccgc ccctaactcc gcccagttcc
gcccattctc cgccccatgg 3120ctgactaatt ttttttattt atgcagaggc
cgaggccgcc tctgcctctg agctattcca 3180gaagtagtga ggaggctttt
ttggaggcct aggcttttgc aaaaagctcc cgggagcttg 3240tatatccatt
ttcggatctg atcaagagac aggatgagga tcgtttcgca tgattgaaca
3300agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg
gctatgactg 3360ggcacaacag acaatcggct gctctgatgc cgccgtgttc
cggctgtcag cgcaggggcg 3420cccggttctt tttgtcaaga ccgacctgtc
cggtgccctg aatgaactgc aggacgaggc 3480agcgcggcta tcgtggctgg
ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt 3540cactgaagcg
ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc
3600atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc
ggcggctgca 3660tacgcttgat ccggctacct gcccattcga ccaccaagcg
aaacatcgca tcgagcgagc 3720acgtactcgg atggaagccg gtcttgtcga
tcaggatgat ctggacgaag agcatcaggg 3780gctcgcgcca gccgaactgt
tcgccaggct caaggcgcgc atgcccgacg gcgaggatct 3840cgtcgtgacc
catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc
3900tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca
tagcgttggc 3960tacccgtgat attgctgaag agcttggcgg cgaatgggct
gaccgcttcc tcgtgcttta 4020cggtatcgcc gctcccgatt cgcagcgcat
cgccttctat cgccttcttg acgagttctt 4080ctgagcggga ctctggggtt
cgaaatgacc gaccaagcga cgcccaacct gccatcacga 4140gatttcgatt
ccaccgccgc cttctatgaa aggttgggct tcggaatcgt tttccgggac
4200gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc
ccaccccaac 4260ttgtttattg cagcttataa tggttacaaa taaagcaata
gcatcacaaa tttcacaaat 4320aaagcatttt tttcactgca ttctagttgt
ggtttgtcca aactcatcaa tgtatcttat 4380catgtctgta taccgtcgac
ctctagctag agcttggcgt aatcatggtc atagctgttt 4440cctgtgtgaa
attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag
4500tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
gcgctcactg 4560cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt
aatgaatcgg ccaacgcgcg 4620gggagaggcg gtttgcgtat tgggcgctct
tccgcttcct cgctcactga ctcgctgcgc 4680tcggtcgttc ggctgcggcg
agcggtatca gctcactcaa aggcggtaat acggttatcc 4740acagaatcag
gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
4800aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
tgacgagcat 4860cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
caggactata aagataccag 4920gcgtttcccc ctggaagctc cctcgtgcgc
tctcctgttc cgaccctgcc gcttaccgga 4980tacctgtccg cctttctccc
ttcgggaagc gtggcgcttt ctcaatgctc acgctgtagg 5040tatctcagtt
cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
5100cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
ggtaagacac 5160gacttatcgc cactggcagc agccactggt aacaggatta
gcagagcgag gtatgtaggc 5220ggtgctacag agttcttgaa gtggtggcct
aactacggct acactagaag gacagtattt 5280ggtatctgcg ctctgctgaa
gccagttacc ttcggaaaaa gagttggtag ctcttgatcc 5340ggcaaacaaa
ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
5400agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
cgctcagtgg 5460aacgaaaact cacgttaagg gattttggtc atgagattat
caaaaaggat cttcacctag 5520atccttttaa attaaaaatg aagttttaaa
tcaatctaaa gtatatatga gtaaacttgg 5580tctgacagtt accaatgctt
aatcagtgag gcacctatct cagcgatctg tctatttcgt 5640tcatccatag
ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
5700tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
agatttatca 5760gcaataaacc agccagccgg aagggccgag cgcagaagtg
gtcctgcaac tttatccgcc 5820tccatccagt ctattaattg ttgccgggaa
gctagagtaa gtagttcgcc agttaatagt 5880ttgcgcaacg ttgttgccat
tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg 5940gcttcattca
gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
6000aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
ggccgcagtg 6060ttatcactca tggttatggc agcactgcat aattctctta
ctgtcatgcc atccgtaaga 6120tgcttttctg tgactggtga gtactcaacc
aagtcattct gagaatagtg tatgcggcga 6180ccgagttgct cttgcccggc
gtcaatacgg gataataccg cgccacatag cagaacttta 6240aaagtgctca
tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
6300ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
atcttttact 6360ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
atgccgcaaa aaagggaata 6420agggcgacac ggaaatgttg aatactcata
ctcttccttt ttcaatatta ttgaagcatt 6480tatcagggtt attgtctcat
gagcggatac atatttgaat gtatttagaa aaataaacaa 6540ataggggttc
cgcgcacatt tccccgaaaa gtgccacctg acgtcgacgg atcgggagat
6600ctcccgatcc cctatggtcg actctcagta caatctgctc tgatgccgca
tagttaagcc 6660agtatctgct ccctgcttgt gtgttggagg tcgctgagta
gtgcgcgagc aaaatttaag 6720ctacaacaag gcaaggcttg accgac
6746415627DNAartificialIgG Lambda Chain Expression Vector
pMORPH?_h_Ig_lambda 41aattgcatga agaatctgct tagggttagg cgttttgcgc
tgcttcgcga tgtacgggcc 60agatatacgc gttgacattg attattgact agttattaat
agtaatcaat tacggggtca 120ttagttcata gcccatatat ggagttccgc
gttacataac ttacggtaaa tggcccgcct 180ggctgaccgc ccaacgaccc
ccgcccattg acgtcaataa tgacgtatgt tcccatagta 240acgccaatag
ggactttcca ttgacgtcaa tgggtggact atttacggta aactgcccac
300ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt
caatgacggt 360aaatggcccg cctggcatta tgcccagtac atgaccttat
gggactttcc tacttggcag 420tacatctacg tattagtcat cgctattacc
atggtgatgc ggttttggca gtacatcaat 480gggcgtggat agcggtttga
ctcacgggga tttccaagtc tccaccccat tgacgtcaat 540gggagtttgt
tttggcacca aaatcaacgg gactttccaa aatgtcgtaa caactccgcc
600ccattgacgc aaatgggcgg taggcgtgta cggtgggagg tctatataag
cagagctctc 660tggctaacta gagaacccac tgcttactgg cttatcgaaa
ttaatacgac tcactatagg 720gagacccaag ctggctagcg ccaccatggc
ctgggctctg ctgctcctca ccctcctcac 780tcagggcaca ggatcctggg
ctgatatcgt gctgacccag ccgccttcag tgagtggcgc 840accaggtcag
cgtgtgacca tctcgtgtag cggcagcagc agcaacattg gtaataatta
900tgtgtcttgg taccagcagt tgcccgggac ggcgccgaaa cttctgattt
ctcgtaattc 960taagcgtccc tcaggcgtgc cggatcgttt tagcggatcc
aaaagcggca ccagcgcgag 1020ccttgcgatt acgggcctgc aaagcgaaga
cgaagcggat tattattgct ctacttatga 1080tactttttct attgtgtttg
gcggcggcac gaagttaacc gtcctaggtc agcccaaggc 1140tgccccctcg
gtcactctgt tcccgccctc ctctgaggag cttcaagcca acaaggccac
1200actggtgtgt ctcataagtg acttctaccc gggagccgtg acagtggcct
ggaagggaga 1260tagcagcccc gtcaaggcgg gagtggagac caccacaccc
tccaaacaaa gcaacaacaa 1320gtacgcggcc agcagctatc tgagcctgac
gcctgagcag tggaagtccc acagaagcta 1380cagctgccag gtcacgcatg
aagggagcac cgtggagaag acagtggccc ctacagaatg 1440ttcatagggg
cccgtttaaa cccgctgatc agcctcgact gtgccttcta gttgccagcc
1500atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca
ctcccactgt 1560cctttcctaa taaaatgagg aaattgcatc gcattgtctg
agtaggtgtc attctattct 1620ggggggtggg gtggggcagg acagcaaggg
ggaggattgg gaagacaata gcaggcatgc 1680tggggatgcg gtgggctcta
tggcttctga ggcggaaaga accagctggg gctctagggg 1740gtatccccac
gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag
1800cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct
tcccttcctt 1860tctcgccacg ttcgccggct ttccccgtca agctctaaat
cggggcatcc ctttagggtt 1920ccgatttagt gctttacggc acctcgaccc
caaaaaactt gattagggtg atggttcacg 1980tagtgggcca tcgccctgat
agacggtttt tcgccctttg acgttggagt ccacgttctt 2040taatagtgga
ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt
2100tgatttataa gggattttgg ggatttcggc ctattggtta aaaaatgagc
tgatttaaca 2160aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt
tagggtgtgg aaagtcccca 2220ggctccccag gcaggcagaa gtatgcaaag
catgcatctc aattagtcag caaccaggtg 2280tggaaagtcc ccaggctccc
cagcaggcag aagtatgcaa agcatgcatc tcaattagtc 2340agcaaccata
gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc
2400ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg
aggccgcctc 2460tgcctctgag ctattccaga agtagtgagg aggctttttt
ggaggcctag gcttttgcaa 2520aaagctcccg ggagcttgta tatccatttt
cggatctgat cagcacgtgt tgacaattaa 2580tcatcggcat agtatatcgg
catagtataa tacgacaagg tgaggaacta aaccatggcc 2640aagttgacca
gtgccgttcc ggtgctcacc gcgcgcgacg tcgccggagc ggtcgagttc
2700tggaccgacc ggctcgggtt ctcccgggac ttcgtggagg acgacttcgc
cggtgtggtc 2760cgggacgacg tgaccctgtt catcagcgcg gtccaggacc
aggtggtgcc ggacaacacc 2820ctggcctggg tgtgggtgcg cggcctggac
gagctgtacg ccgagtggtc ggaggtcgtg 2880tccacgaact tccgggacgc
ctccgggccg gccatgaccg agatcggcga gcagccgtgg 2940gggcgggagt
tcgccctgcg cgacccggcc ggcaactgcg tgcacttcgt ggccgaggag
3000caggactgac acgtgctacg agatttcgat tccaccgccg ccttctatga
aaggttgggc 3060ttcggaatcg ttttccggga cgccggctgg atgatcctcc
agcgcgggga tctcatgctg 3120gagttcttcg cccaccccaa cttgtttatt
gcagcttata atggttacaa ataaagcaat 3180agcatcacaa atttcacaaa
taaagcattt ttttcactgc attctagttg tggtttgtcc 3240aaactcatca
atgtatctta tcatgtctgt ataccgtcga cctctagcta gagcttggcg
3300taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat
tccacacaac 3360atacgagccg gaagcataaa gtgtaaagcc tggggtgcct
aatgagtgag ctaactcaca 3420ttaattgcgt tgcgctcact gcccgctttc
cagtcgggaa acctgtcgtg ccagctgcat 3480taatgaatcg gccaacgcgc
ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc 3540tcgctcactg
actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
3600aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
catgtgagca 3660aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
tgctggcgtt tttccatagg 3720ctccgccccc ctgacgagca tcacaaaaat
cgacgctcaa gtcagaggtg gcgaaacccg 3780acaggactat aaagatacca
ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt 3840ccgaccctgc
cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
3900tctcaatgct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
caagctgggc 3960tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
tatccggtaa ctatcgtctt 4020gagtccaacc cggtaagaca cgacttatcg
ccactggcag cagccactgg taacaggatt 4080agcagagcga ggtatgtagg
cggtgctaca gagttcttga agtggtggcc taactacggc 4140tacactagaa
ggacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
4200agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
tttttttgtt 4260tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
aagatccttt gatcttttct 4320acggggtctg acgctcagtg gaacgaaaac
tcacgttaag ggattttggt catgagatta 4380tcaaaaagga tcttcaccta
gatcctttta aattaaaaat gaagttttaa atcaatctaa 4440agtatatatg
agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
4500tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
gtagataact 4560acgatacggg agggcttacc atctggcccc agtgctgcaa
tgataccgcg agacccacgc 4620tcaccggctc cagatttatc agcaataaac
cagccagccg gaagggccga gcgcagaagt 4680ggtcctgcaa ctttatccgc
ctccatccag tctattaatt gttgccggga agctagagta 4740agtagttcgc
cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
4800tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
aaggcgagtt 4860acatgatccc ccatgttgtg caaaaaagcg gttagctcct
tcggtcctcc gatcgttgtc 4920agaagtaagt tggccgcagt gttatcactc
atggttatgg cagcactgca taattctctt 4980actgtcatgc catccgtaag
atgcttttct gtgactggtg agtactcaac caagtcattc 5040tgagaatagt
gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
5100gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
ggggcgaaaa 5160ctctcaagga tcttaccgct gttgagatcc agttcgatgt
aacccactcg tgcacccaac 5220tgatcttcag catcttttac tttcaccagc
gtttctgggt gagcaaaaac aggaaggcaa 5280aatgccgcaa aaaagggaat
aagggcgaca cggaaatgtt gaatactcat actcttcctt 5340tttcaatatt
attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
5400tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
agtgccacct 5460gacgtcgacg gatcgggaga tctcccgatc ccctatggtc
gactctcagt acaatctgct 5520ctgatgccgc atagttaagc cagtatctgc
tccctgcttg tgtgttggag gtcgctgagt 5580agtgcgcgag caaaatttaa
gctacaacaa ggcaaggctt gaccgac 5627425629DNAartificialIgG Kappa
Chain Expression Vector pMORPH?_h_Ig_kappa 42aattgcatga agaatctgct
tagggttagg cgttttgcgc tgcttcgcga tgtacgggcc 60agatatacgc gttgacattg
attattgact agttattaat agtaatcaat tacggggtca 120ttagttcata
gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct
180ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt
tcccatagta 240acgccaatag ggactttcca ttgacgtcaa tgggtggagt
atttacggta aactgcccac 300ttggcagtac atcaagtgta tcatatgcca
agtacgcccc ctattgacgt caatgacggt 360aaatggcccg cctggcatta
tgcccagtac atgaccttat gggactttcc tacttggcag 420tacatctacg
tattagtcat cgctattacc atggtgatgc ggttttggca gtacatcaat
480gggcgtggat agcggtttga ctcacgggga tttccaagtc tccaccccat
tgacgtcaat 540gggagtttgt tttggcacca aaatcaacgg gactttccaa
aatgtcgtaa caactccgcc 600ccattgacgc aaatgggcgg taggcgtgta
cggtgggagg tctatataag cagagctctc 660tggctaacta gagaacccac
tgcttactgg cttatcgaaa ttaatacgac tcactatagg 720gagacccaag
ctggctagcg ccaccatggt gttgcagacc caggtcttca tttctctgtt
780gctctggatc tctggtgcct acggggatat ccagatgacc cagagcccgt
ctagcctgag 840cgcgagcgtg ggtgatcgtg tgaccattac ctgcagagcg
agccagtcta tttctaattg 900gctgaattgg taccagcaga aaccaggtaa
agcaccgaaa ctattaattt ataaggcttc 960tactttgcaa agcggggtcc
cgtcccgttt tagcggctct ggatccggca ctgattttac 1020cctgaccatt
agcagcctgc aacctgaaga ctttgcgact tattattgcc agcagtatgg
1080taatattcct attacctttg gccagggtac gaaagttgaa attaaacgta
cggtggctgc 1140accatctgtc ttcatcttcc cgccatctga tgagcagttg
aaatctggaa ctgcctctgt 1200tgtgtgcctg ctgaataact tctatcccag
agaggccaaa gtacagtgga aggtggataa 1260cgccctccaa tcgggtaact
cccaggagag tgtcacagag caggacagca aggacagcac 1320ctacagcctc
agcagcaccc tgacgctgag caaagcagac tacgagaaac acaaagtcta
1380cgcctgcgaa gtcacccatc agggcctgag ctcgcccgtc acaaagagct
tcaacagggg 1440agagtgttag gggcccgttt aaacccgctg atcagcctcg
actgtgcctt ctagttgcca 1500gccatctgtt gtttgcccct cccccgtgcc
ttccttgacc ctggaaggtg ccactcccac 1560tgtcctttcc taataaaatg
aggaaattgc atcgcattgt ctgagtaggt gtcattctat 1620tctggggggt
ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca
1680tgctggggat gcggtgggct ctatggcttc tgaggcggaa agaaccagct
ggggctctag 1740ggggtatccc cacgcgccct gtagcggcgc attaagcgcg
gcgggtgtgg tggttacgcg 1800cagcgtgacc gctacacttg ccagcgccct
agcgcccgct cctttcgctt tcttcccttc 1860ctttctcgcc acgttcgccg
gctttccccg tcaagctcta aatcgggggc tccctttagg 1920gttccgattt
agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc
1980acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg
agtccacgtt 2040ctttaatagt ggactcttgt tccaaactgg aacaacactc
aaccctatct cggtctattc 2100ttttgattta taagggattt tgccgatttc
ggcctattgg ttaaaaaatg agctgattta 2160acaaaaattt aacgcgaatt
aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc 2220ccaggctccc
cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg
2280tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
tctcaattag 2340tcagcaacca tagtcccgcc cctaactccg cccatcccgc
ccctaactcc gcccagttcc 2400gcccattctc cgccccatgg ctgactaatt
ttttttattt atgcagaggc cgaggccgcc 2460tctgcctctg agctattcca
gaagtagtga ggaggctttt ttggaggcct aggcttttgc 2520aaaaagctcc
cgggagcttg tatatccatt ttcggatctg atcagcacgt gttgacaatt
2580aatcatcggc atagtatatc ggcatagtat aatacgacaa ggtgaggaac
taaaccatgg 2640ccaagttgac cagtgccgtt ccggtgctca ccgcgcgcga
cgtcgccgga gcggtcgagt 2700tctggaccga ccggctcggg ttctcccggg
acttcgtgga ggacgacttc gccggtgtgg 2760tccgggacga cgtgaccctg
ttcatcagcg cggtccagga ccaggtggtg ccggacaaca 2820ccctggcctg
ggtgtgggtg cgcggcctgg acgagctgta cgccgagtgg tcggaggtcg
2880tgtccacgaa cttccgggac gcctccgggc cggccatgac cgagatcggc
gagcagccgt 2940gggggcggga gttcgccctg cgcgacccgg ccggcaactg
cgtgcacttc gtggccgagg 3000agcaggactg acacgtgcta cgagatttcg
attccaccgc cgccttctat gaaaggttgg 3060gcttcggaat cgttttccgg
gacgccggct ggatgatcct ccagcgcggg gatctcatgc 3120tggagttctt
cgcccacccc aacttgttta ttgcagctta taatggttac aaataaagca
3180atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt
tgtggtttgt 3240ccaaactcat caatgtatct tatcatgtct gtataccgtc
gacctctagc tagagcttgg 3300cgtaatcatg gtcatagctg tttcctgtgt
gaaattgtta tccgctcaca attccacaca 3360acatacgagc cggaagcata
aagtgtaaag cctggggtgc ctaatgagtg agctaactca 3420cattaattgc
gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc
3480attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc
tcttccgctt 3540cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
gcgagcggta tcagctcact 3600caaaggcggt aatacggtta tccacagaat
caggggataa cgcaggaaag aacatgtgag 3660caaaaggcca gcaaaaggcc
aggaaccgta aaaaggccgc gttgctggcg tttttccata 3720ggctccgccc
ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
3780cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
cgctctcctg 3840ttccgaccct gccgcttacc ggatacctgt ccgcctttct
cccttcggga agcgtggcgc 3900tttctcatag ctcacgctgt aggtatctca
gttcggtgta ggtcgttcgc tccaagctgg 3960gctgtgtgca cgaacccccc
gttcagcccg accgctgcgc cttatccggt aactatcgtc 4020ttgagtccaa
cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
4080ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
cctaactacg 4140gctacactag aagaacagta tttggtatct gcgctctgct
gaagccagtt accttcggaa 4200aaagagttgg tagctcttga tccggcaaac
aaaccaccgc tggtagcggt ggtttttttg 4260tttgcaagca gcagattacg
cgcagaaaaa aaggatctca agaagatcct ttgatctttt 4320ctacggggtc
tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
4380tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
aaatcaatct 4440aaagtatata tgagtaaact tggtctgaca gttaccaatg
cttaatcagt gaggcaccta 4500tctcagcgat ctgtctattt cgttcatcca
tagttgcctg actccccgtc gtgtagataa 4560ctacgatacg ggagggctta
ccatctggcc ccagtgctgc aatgataccg cgagacccac 4620gctcaccggc
tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
4680gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
gaagctagag 4740taagtagttc gccagttaat agtttgcgca acgttgttgc
cattgctaca ggcatcgtgg 4800tgtcacgctc gtcgtttggt atggcttcat
tcagctccgg ttcccaacga tcaaggcgag 4860ttacatgatc ccccatgttg
tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg 4920tcagaagtaa
gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
4980ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
accaagtcat 5040tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
ggcgtcaata cgggataata 5100ccgcgccaca tagcagaact ttaaaagtgc
tcatcattgg aaaacgttct tcggggcgaa 5160aactctcaag gatcttaccg
ctgttgagat ccagttcgat gtaacccact cgtgcaccca 5220actgatcttc
agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
5280aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
atactcttcc 5340tttttcaata ttattgaagc atttatcagg gttattgtct
catgagcgga tacatatttg 5400aatgtattta gaaaaataaa caaatagggg
ttccgcgcac atttccccga aaagtgccac 5460ctgacgtcga cggatcggga
gatctcccga tcccctatgg tgcactctca gtacaatctg 5520ctctgatgcc
gcatagttaa gccagtatct gctccctgct tgtgtgttgg aggtcgctga
5580gtagtgcgcg agcaaaattt aagctacaac aaggcaaggc ttgaccgac
5629437101DNAartificialIgG4 Expression Vector pMORPH2?_h_Ig 4
43taatacgact cactataggg agacccaagc tggctagcgc caccatgaaa cacctgtggt
60tcttcctcct gctggtggca gctcccagat gggtcctgtc ccaggtgcaa ttgcaacagt
120ctggtccggg cctggtgaaa ccgagccaaa ccctgagcct gacctgtgcg
atttccggag 180atagcgtgag cgataattct gctgcttggt cttggattcg
ccagtctcct gggcgtggcc 240tcgagtggct gggccttatc tatcttcgta
gcaagtggga taacgattat gcggtgagcg 300tgaaaagccg gattaccatc
aacccggata cttcgaaaaa ccagtttagc ctgcaactga 360acagcgtgac
cccggaagat acggccgtgt attattgcgc gcgtactggt cgtgctgatg
420agtttgatgt ttggggccaa ggcaccctgg tgacggttag ctcagcttcc
accaagggac 480catccgtctt ccccctggcg ccctgctcca ggagcacctc
cgagagcaca gccgccctgg 540gctgcctggt caaggactac ttccccgaac
cggtgacggt gtcgtggaac tcaggcgccc 600tgaccagcgg cgtgcacacc
ttcccggctg tcctacagtc ctcaggactc tactccctca 660gcagcgtggt
gaccgtgccc tccagcagct tgggcacgaa gacctacacc tgcaacgtag
720atcacaagcc cagcaacacc aaggtggaca agagagttga gtccaaatat
ggtcccccat 780gcccatcatg cccagcacct gagttcctgg ggggaccatc
agtcttcctg ttccccccaa 840aacccaagga cactctcatg atctcccgga
cccctgaggt cacgtgcgtg gtggtggacg 900tgagccagga agaccccgag
gtccagttca actggtacgt ggatggcgtg gaggtgcata 960atgccaagac
aaagccgcgg gaggagcagt tcaacagcac gtaccgtgtg gtcagcgtcc
1020tcaccgtcct gcaccaggac tggctgaacg gcaaggagta caagtgcaag
gtctccaaca 1080aaggcctccc gtcctccatc gagaaaacca tctccaaagc
caaagggcag ccccgagagc 1140cacaggtgta caccctgccc ccatcccagg
aggagatgac caagaaccag gtcagcctga 1200cctgcctggt caaaggcttc
taccccagcg acatcgccgt ggagtgggag agcaatgggc 1260agccggagaa
caactacaag accacgcctc ccgtgctgga ctccgacggc tccttcttcc
1320tctacagcag gctaaccgtg gacaagagca ggtggcagga ggggaatgtc
ttctcatgct 1380ccgtgatgca tgaggctctg cacaaccact acacacagaa
gagcctctcc ctgtctctgg 1440gtaaatgagg gcccgtttaa acgggtggca
tccctgtgac ccctccccag tgcctctcct 1500ggccctggaa gttgccactc
cagtgcccac cagccttgtc ctaataaaat taagttgcat 1560cattttgtct
gactaggtgt ccttctataa tattatgggg tggagggggg tggtatggag
1620caaggggcaa gttgggaaga caacctgtag ggcctgcggg gtctattggg
aaccaagctg 1680gagtgcagtg gcacaatctt ggctcactgc aatctccgcc
tcctgggttc aagcgattct 1740cctgcctcag cctcccgagt tgttgggatt
ccaggcatgc atgaccaggc tcacctaatt 1800tttgtttttt tggtagagac
ggggtttcac catattggcc aggctggtct ccaactccta 1860atctcaggtg
atctacccac cttggcctcc caaattgctg ggattacagg cgtgaaccac
1920tgctcccttc cctgtccttc tgattttaaa ataactatac cagcaggagg
acgtccagac 1980acagcatagg ctacctggcc atgcccaacc ggtgggacat
ttgagttgct tgcttggcac 2040tgtcctctca tgcgttgggt ccactcagta
gatgcctgtt gaattgggta cgcggcatcg 2100attccacgcg ccctgtagcg
gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt 2160gaccgctaca
cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct
2220cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt
tagggttccg 2280atttagtgct ttacggcacc tcgaccccaa aaaacttgat
tagggtgatg gttcacgtag 2340tgggccatcg ccctgataga cggtttttcg
ccctttgacg ttggagtcca cgttctttaa 2400tagtggactc ttgttccaaa
ctggaacaac actcaaccct atctcggtct attcttttga 2460tttataaggg
attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa
2520atttaacgcg aattaattct gtggaatgtg tgtcagttag ggtgtggaaa
gtccccaggc 2580tccccagcag gcagaagtat gcaaagcatg catctcaatt
agtcagcaac caggtgtgga 2640aagtccccag gctccccagc aggcagaagt
atgcaaagca tgcatctcaa ttagtcagca 2700accatagtcc cgcccctaac
tccgcccatc ccgcccctaa ctccgcccag ttccgcccat 2760tctccgcccc
atggctgact aatttttttt atttatgcag aggccgaggc cgcctctgcc
2820tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt
ttgcaaaaag 2880ctcccgggag cttgtatatc cattttcgga tctgatcaag
agacaggatg aggatcgttt 2940cgcatgattg aacaagatgg attgcacgca
ggttctccgg ccgcttgggt ggagaggcta 3000ttcggctatg actgggcaca
acagacaatc ggctgctctg atgccgccgt gttccggctg 3060tcagcgcagg
ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa
3120ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc
ttgcgcagct 3180gtgctcgacg ttgtcactga agcgggaagg gactggctgc
tattgggcga agtgccgggg 3240caggatctcc tgtcatctca ccttgctcct
gccgagaaag tatccatcat ggctgatgca 3300atgcggcggc tgcatacgct
tgatccggct acctgcccat tcgaccacca agcgaaacat 3360cgcatcgagc
gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac
3420gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc
gcgcatgccc 3480gacggcgagg atctcgtcgt gacccatggc gatgcctgct
tgccgaatat catggtggaa 3540aatggccgct tttctggatt catcgactgt
ggccggctgg gtgtggcgga ccgctatcag 3600gacatagcgt tggctacccg
tgatattgct gaagagcttg gcggcgaatg ggctgaccgc 3660ttcctcgtgc
tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt
3720cttgacgagt tcttctgagc gggactctgg ggttcgaaat gaccgaccaa
gcgacgccca 3780acctgccatc acgagatttc gattccaccg ccgccttcta
tgaaaggttg ggcttcggaa 3840tcgttttccg ggacgccggc tggatgatcc
tccagcgcgg ggatctcatg ctggagttct 3900tcgcccaccc caacttgttt
attgcagctt ataatggtta caaataaagc aatagcatca 3960caaatttcac
aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca
4020tcaatgtatc ttatcatgtc tgtataccgt cgacctctag ctagagcttg
gcgtaatcat 4080ggtcatagct gtttcctgtg tgaaattgtt atccgctcac
aattccacac aacatacgag 4140ccggaagcat aaagtgtaaa gcctggggtg
cctaatgagt gagctaactc acattaattg 4200cgttgcgctc actgcccgct
ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa 4260tcggccaacg
cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca
4320ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
tcaaaggcgg 4380taatacggtt atccacagaa tcaggggata acgcaggaaa
gaacatgtga gcaaaaggcc 4440agcaaaaggc caggaaccgt aaaaaggccg
cgttgctggc gtttttccat aggctccgcc 4500cccctgacga gcatcacaaa
aatcgacgct caagtcagag gtggcgaaac ccgacaggac 4560tataaagata
ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
4620tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
ctttctcata 4680gctcacgctg taggtatctc agttcggtgt aggtcgttcg
ctccaagctg ggctgtgtgc 4740acgaaccccc cgttcagccc gaccgctgcg
ccttatccgg taactatcgt cttgagtcca 4800acccggtaag acacgactta
tcgccactgg cagcagccac tggtaacagg attagcagag 4860cgaggtatgt
aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
4920gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
aaaagagttg 4980gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
tggttttttt gtttgcaagc 5040agcagattac gcgcagaaaa aaaggatctc
aagaagatcc tttgatcttt tctacggggt 5100ctgacgctca gtggaacgaa
aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa 5160ggatcttcac
ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
5220atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
atctcagcga 5280tctgtctatt tcgttcatcc atagttgcct gactccccgt
cgtgtagata actacgatac 5340gggagggctt accatctggc cccagtgctg
caatgatacc gcgagaccca cgctcaccgg 5400ctccagattt atcagcaata
aaccagccag ccggaagggc cgagcgcaga agtggtcctg 5460caactttatc
cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
5520cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
gtgtcacgct 5580cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
atcaaggcga gttacatgat 5640cccccatgtt gtgcaaaaaa gcggttagct
ccttcggtcc tccgatcgtt gtcagaagta 5700agttggccgc agtgttatca
ctcatggtta tggcagcact gcataattct cttactgtca 5760tgccatccgt
aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
5820agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
accgcgccac 5880atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
ttcggggcga aaactctcaa 5940ggatcttacc gctgttgaga tccagttcga
tgtaacccac tcgtgcaccc aactgatctt 6000cagcatcttt tactttcacc
agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg 6060caaaaaaggg
aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
6120attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
gaatgtattt 6180agaaaaataa acaaataggg gttccgcgca catttccccg
aaaagtgcca cctgacgtcg 6240acggatcggg agatctcccg atcccctatg
gtgcactctc agtacaatct gctctgatgc 6300cgcatagtta agccagtatc
tgctccctgc ttgtgtgttg gaggtcgctg agtagtgcgc 6360gagcaaaatt
taagctacaa caaggcaagg cttgaccgac atttgcatga agaatctgct
6420tagggttagg cgttttgcgc tgcttcgcga tgtacgggcc agatatacgc
gttgacattg 6480attattgact agttattaat agtaatcaat tacggggtca
ttagttcata gcccatatat 6540ggagttccgc gttacataac ttacggtaaa
tggcccgcct ggctgaccgc ccaacgaccc 6600ccgcccattg acgtcaataa
tgacgtatgt tcccatagta acgccaatag ggactttcca 6660ttgacgtcaa
tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta
6720tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
cctggcatta 6780tgcccagtac atgaccttat gggactttcc tacttggcag
tacatctacg tattagtcat 6840cgctattacc atggtgatgc ggttttggca
gtacatcaat gggcgtggat agcggtttga 6900ctcacgggga tttccaagtc
tccaccccat tgacgtcaat gggagtttgt tttggcacca 6960aaatcaacgg
gactttccaa aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg
7020taggcgtgta cggtgggagg tctatataag cagagctctc
tggctaacta gagaacccac 7080tgcttactgg cttatcgaaa t
7101445983DNAartificialIgG Lambda Chain Expression Vector
pMORPH?2_h_Ig_lambda2 44taatacgact cactataggg agacccaagc tggctagcgc
caccatggcc tgggctctgc 60tgctcctcac cctcctcact cagggcacag gatcctgggc
tgatatcgtg ctgacccagc 120cgccttcagt gagtggcgca ccaggtcagc
gtgtgaccat ctcgtgtagc ggcagcagca 180gcaacattgg taataattat
gtgtcttggt accagcagtt gcccgggacg gcgccgaaac 240ttctgatttc
tcgtaattct aagcgtccct caggcgtgcc ggatcgtttt agcggatcca
300aaagcggcac cagcgcgagc cttgcgatta cgggcctgca aagcgaagac
gaagcggatt 360attattgctc tacttatgat actttttcta ttgtgtttgg
cggcggcacg aagttaaccg 420tcctaggtca gcccaaggct gccccctcgg
tcactctgtt cccgccctcc tctgaggagc 480ttcaagccaa caaggccaca
ctggtgtgtc tcataagtga cttctacccg ggagccgtga 540cagtggcctg
gaaggcagat agcagccccg tcaaggcggg agtggagacc accacaccct
600ccaaacaaag caacaacaag tacgcggcca gcagctatct gagcctgacg
cctgagcagt 660ggaagtccca cagaagctac agctgccagg tcacgcatga
agggagcacc gtggagaaga 720cagtggcccc tacagaatgt tcataggggc
ccgtttaaac gggtggcatc cctgtgaccc 780ctccccagtg cctctcctgg
ccctggaagt tgccactcca gtgcccacca gccttgtcct 840aataaaatta
agttgcatca ttttgtctga ctaggtgtcc ttctataata ttatggggtg
900gaggggggtg gtatggagca aggggcaagt tgggaagaca acctgtaggg
cctgcggggt 960ctattgggaa ccaagctgga gtgcagtggc acaatcttgg
ctcactgcaa tctccgcctc 1020ctgggttcaa gcgattctcc tgcctcagcc
tcccgagttg ttgggattcc aggcatgcat 1080gaccaggctc acctaatttt
tgtttttttg gtagagacgg ggtttcacca tattggccag 1140gctggtctcc
aactcctaat ctcaggtgat ctacccacct tggcctccca aattgctggg
1200attacaggcg tgaaccactg ctcccttccc tgtccttctg attttaaaat
aactatacca 1260gcaggaggac gtccagacac agcataggct acctggccat
gcccaaccgg tgggacattt 1320gagttgcttg cttggcactg tcctctcatg
cgttgggtcc actcagtaga tgcctgttga 1380attgggtacg cggcatcgat
tccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt 1440ggtggttacg
cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc
1500tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc
taaatcgggg 1560gctcccttta gggttccgat ttagtgcttt acggcacctc
gaccccaaaa aacttgatta 1620gggtgatggt tcacgtagtg ggccatcgcc
ctgatagacg gtttttcgcc ctttgacgtt 1680ggagtccacg ttctttaata
gtggactctt gttccaaact ggaacaacac tcaaccctat 1740ctcggtctat
tcttttgatt tataagggat tttggggatt tcggcctatt ggttaaaaaa
1800tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg
tcagttaggg 1860tgtggaaagt ccccaggctc cccaggcagg cagaagtatg
caaagcatgc atctcaatta 1920gtcagcaacc aggtgtggaa agtccccagg
ctccccagca ggcagaagta tgcaaagcat 1980gcatctcaat tagtcagcaa
ccatagtccc gcccctaact ccgcccatcc cgcccctaac 2040tccgcccagt
tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga
2100ggccgaggcc gcctctgcct ctgagctatt ccagaagtag tgaggaggct
tttttggagg 2160cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc
attttcggat ctgatcagca 2220cgtgttgaca attaatcatc ggcatagtat
atcggcatag tataatacga caaggtgagg 2280aactaaacca tggccaagtt
gaccagtgcc gttccggtgc tcaccgcgcg cgacgtcgcc 2340ggagcggtcg
agttctggac cgaccggctc gggttctccc gggacttcgt ggaggacgac
2400ttcgccggtg tggtccggga cgacgtgacc ctgttcatca gcgcggtcca
ggaccaggtg 2460gtgccggaca acaccctggc ctgggtgtgg gtgcgcggcc
tggacgagct gtacgccgag 2520tggtcggagg tcgtgtccac gaacttccgg
gacgcctccg ggccggccat gaccgagatc 2580ggcgagcagc cgtgggggcg
ggagttcgcc ctgcgcgacc cggccggcaa ctgcgtgcac 2640ttcgtggccg
aggagcagga ctgacacgtg ctacgagatt tcgattccac cgccgccttc
2700tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat
cctccagcgc 2760ggggatctca tgctggagtt cttcgcccac cccaacttgt
ttattgcagc ttataatggt 2820tacaaataaa gcaatagcat cacaaatttc
acaaataaag catttttttc actgcattct 2880agttgtggtt tgtccaaact
catcaatgta tcttatcatg tctgtatacc gtcgacctct 2940agctagagct
tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc
3000acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg
tgcctaatga 3060gtgagctaac tcacattaat tgcgttgcgc tcactgcccg
ctttccagtc gggaaacctg 3120tcgtgccagc tgcattaatg aatcggccaa
cgcgcgggga gaggcggttt gcgtattggg 3180cgctcttccg cttcctcgct
cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg 3240gtatcagctc
actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
3300aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
cgcgttgctg 3360gcgtttttcc ataggctccg cccccctgac gagcatcaca
aaaatcgacg ctcaagtcag 3420aggtggcgaa acccgacagg actataaaga
taccaggcgt ttccccctgg aagctccctc 3480gtgcgctctc ctgttccgac
cctgccgctt accggatacc tgtccgcctt tctcccttcg 3540ggaagcgtgg
cgctttctca atgctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
3600cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
cgccttatcc 3660ggtaactatc gtcttgagtc caacccggta agacacgact
tatcgccact ggcagcagcc 3720actggtaaca ggattagcag agcgaggtat
gtaggcggtg ctacagagtt cttgaagtgg 3780tggcctaact acggctacac
tagaaggaca gtatttggta tctgcgctct gctgaagcca 3840gttaccttcg
gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
3900ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
tcaagaagat 3960cctttgatct tttctacggg gtctgacgct cagtggaacg
aaaactcacg ttaagggatt 4020ttggtcatga gattatcaaa aaggatcttc
acctagatcc ttttaaatta aaaatgaagt 4080tttaaatcaa tctaaagtat
atatgagtaa acttggtctg acagttacca atgcttaatc 4140agtgaggcac
ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
4200gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
tgcaatgata 4260ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
taaaccagcc agccggaagg 4320gccgagcgca gaagtggtcc tgcaacttta
tccgcctcca tccagtctat taattgttgc 4380cgggaagcta gagtaagtag
ttcgccagtt aatagtttgc gcaacgttgt tgccattgct 4440acaggcatcg
tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
4500cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
ctccttcggt 4560cctccgatcg ttgtcagaag taagttggcc gcagtgttat
cactcatggt tatggcagca 4620ctgcataatt ctcttactgt catgccatcc
gtaagatgct tttctgtgac tggtgagtac 4680tcaaccaagt cattctgaga
atagtgtatg cggcgaccga gttgctcttg cccggcgtca 4740atacgggata
ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
4800tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
gatgtaaccc 4860actcgtgcac ccaactgatc ttcagcatct tttactttca
ccagcgtttc tgggtgagca 4920aaaacaggaa ggcaaaatgc cgcaaaaaag
ggaataaggg cgacacggaa atgttgaata 4980ctcatactct tcctttttca
atattattga agcatttatc agggttattg tctcatgagc 5040ggatacatat
ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
5100cgaaaagtgc cacctgacgt cgacggatcg ggagatctcc cgatccccta
tggtcgactc 5160tcagtacaat ctgctctgat gccgcatagt taagccagta
tctgctccct gcttgtgtgt 5220tggaggtcgc tgagtagtgc gcgagcaaaa
tttaagctac aacaaggcaa ggcttgaccg 5280acaattgcat gaagaatctg
cttagggtta ggcgttttgc gctgcttcgc gatgtacggg 5340ccagatatac
gcgttgacat tgattattga ctagttatta atagtaatca attacggggt
5400cattagttca tagcccatat atggagttcc gcgttacata acttacggta
aatggcccgc 5460ctggctgacc gcccaacgac ccccgcccat tgacgtcaat
aatgacgtat gttcccatag 5520taacgccaat agggactttc cattgacgtc
aatgggtgga ctatttacgg taaactgccc 5580acttggcagt acatcaagtg
tatcatatgc caagtacgcc ccctattgac gtcaatgacg 5640gtaaatggcc
cgcctggcat tatgcccagt acatgacctt atgggacttt cctacttggc
5700agtacatcta cgtattagtc atcgctatta ccatggtgat gcggttttgg
cagtacatca 5760atgggcgtgg atagcggttt gactcacggg gatttccaag
tctccacccc attgacgtca 5820atgggagttt gttttggcac caaaatcaac
gggactttcc aaaatgtcgt aacaactccg 5880ccccattgac gcaaatgggc
ggtaggcgtg tacggtggga ggtctatata agcagagctc 5940tctggctaac
tagagaaccc actgcttact ggcttatcga aat 5983455986DNAartificialIgG
kappa Chain Expression Vector pMORPH?2_h_Ig_kappa 45taatacgact
cactataggg agacccaagc tggctagcgc caccatggtg ttgcagaccc 60aggtcttcat
ttctctgttg ctctggatct ctggtgccta cggggatatc cagatgaccc
120agagcccgtc tagcctgagc gcgagcgtgg gtgatcgtgt gaccattacc
tgcagagcga 180gccagtctat ttctaattgg ctgaattggt accagcagaa
accaggtaaa gcaccgaaac 240tattaattta taaggcttct actttgcaaa
gcggggtccc gtcccgtttt agcggctctg 300gatccggcac tgattttacc
ctgaccatta gcagcctgca acctgaagac tttgcgactt 360attattgcca
gcagtatggt aatattccta ttacctttgg ccagggtacg aaagttgaaa
420ttaaacgtac ggtggctgca ccatctgtct tcatcttccc gccatctgat
gagcagttga 480aatctggaac tgcctctgtt gtgtgcctgc tgaataactt
ctatcccaga gaggccaaag 540tacagtggaa ggtggataac gccctccaat
cgggtaactc ccaggagagt gtcacagagc 600aggacagcaa ggacagcacc
tacagcctca gcagcaccct gacgctgagc aaagcagact 660acgagaaaca
caaagtctac gcctgcgaag tcacccatca gggcctgagc tcgcccgtca
720caaagagctt caacagggga gagtgttagg ggcccgttta aacgggtggc
atccctgtga 780cccctcccca gtgcctctcc tggccctgga agttgccact
ccagtgccca ccagccttgt 840cctaataaaa ttaagttgca tcattttgtc
tgactaggtg tccttctata atattatggg 900gtggaggggg gtggtatgga
gcaaggggca agttgggaag acaacctgta gggcctgcgg 960ggtctattgg
gaaccaagct ggagtgcagt ggcacaatct tggctcactg caatctccgc
1020ctcctgggtt caagcgattc tcctgcctca gcctcccgag ttgttgggat
tccaggcatg 1080catgaccagg ctcacctaat ttttgttttt ttggtagaga
cggggtttca ccatattggc 1140caggctggtc tccaactcct aatctcaggt
gatctaccca ccttggcctc ccaaattgct 1200gggattacag gcgtgaacca
ctgctccctt ccctgtcctt ctgattttaa aataactata 1260ccagcaggag
gacgtccaga cacagcatag gctacctggc catgcccaac cggtgggaca
1320tttgagttgc ttgcttggca ctgtcctctc atgcgttggg tccactcagt
agatgcctgt 1380tgaattgggt acgcggcatc gattccacgc gccctgtagc
ggcgcattaa gcgcggcggg 1440tgtggtggtt acgcgcagcg tgaccgctac
acttgccagc gccctagcgc ccgctccttt 1500cgctttcttc ccttcctttc
tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg 1560ggggctccct
ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga
1620ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc
gccctttgac 1680gttggagtcc acgttcttta atagtggact cttgttccaa
actggaacaa cactcaaccc 1740tatctcggtc tattcttttg atttataagg
gattttgggg atttcggcct attggttaaa 1800aaatgagctg atttaacaaa
aatttaacgc gaattaattc tgtggaatgt gtgtcagtta 1860gggtgtggaa
agtccccagg ctccccaggc aggcagaagt atgcaaagca tgcatctcaa
1920ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa
gtatgcaaag 1980catgcatctc aattagtcag caaccatagt cccgccccta
actccgccca tcccgcccct 2040aactccgccc agttccgccc attctccgcc
ccatggctga ctaatttttt ttatttatgc 2100agaggccgag gccgcctctg
cctctgagct attccagaag tagtgaggag gcttttttgg 2160aggcctaggc
ttttgcaaaa agctcccggg agcttgtata tccattttcg gatctgatca
2220gcacgtgttg acaattaatc atcggcatag tatatcggca tagtataata
cgacaaggtg 2280aggaactaaa ccatggccaa gttgaccagt gccgttccgg
tgctcaccgc gcgcgacgtc 2340gccggagcgg tcgagttctg gaccgaccgg
ctcgggttct cccgggactt cgtggaggac 2400gacttcgccg gtgtggtccg
ggacgacgtg accctgttca tcagcgcggt ccaggaccag 2460gtggtgccgg
acaacaccct ggcctgggtg tgggtgcgcg gcctggacga gctgtacgcc
2520gagtggtcgg aggtcgtgtc cacgaacttc cgggacgcct ccgggccggc
catgaccgag 2580atcggcgagc agccgtgggg gcgggagttc gccctgcgcg
acccggccgg caactgcgtg 2640cacttcgtgg ccgaggagca ggactgacac
gtgctacgag atttcgattc caccgccgcc 2700ttctatgaaa ggttgggctt
cggaatcgtt ttccgggacg ccggctggat gatcctccag 2760cgcggggatc
tcatgctgga gttcttcgcc caccccaact tgtttattgc agcttataat
2820ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
ttcactgcat 2880tctagttgtg gtttgtccaa actcatcaat gtatcttatc
atgtctgtat accgtcgacc 2940tctagctaga gcttggcgta atcatggtca
tagctgtttc ctgtgtgaaa ttgttatccg 3000ctcacaattc cacacaacat
acgagccgga agcataaagt gtaaagcctg gggtgcctaa 3060tgagtgagct
aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac
3120ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
tttgcgtatt 3180gggcgctctt ccgcttcctc gctcactgac tcgctgcgct
cggtcgttcg gctgcggcga 3240gcggtatcag ctcactcaaa ggcggtaata
cggttatcca cagaatcagg ggataacgca 3300ggaaagaaca tgtgagcaaa
aggccagcaa aaggccagga accgtaaaaa ggccgcgttg 3360ctggcgtttt
tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
3420cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
tggaagctcc 3480ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
acctgtccgc ctttctccct 3540tcgggaagcg tggcgctttc tcaatgctca
cgctgtaggt atctcagttc ggtgtaggtc 3600gttcgctcca agctgggctg
tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta 3660tccggtaact
atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
3720gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
gttcttgaag 3780tggtggccta actacggcta cactagaagg acagtatttg
gtatctgcgc tctgctgaag 3840ccagttacct tcggaaaaag agttggtagc
tcttgatccg gcaaacaaac caccgctggt 3900agcggtggtt tttttgtttg
caagcagcag attacgcgca gaaaaaaagg atctcaagaa 3960gatcctttga
tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
4020attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
ttaaaaatga 4080agttttaaat caatctaaag tatatatgag taaacttggt
ctgacagtta ccaatgctta 4140atcagtgagg cacctatctc agcgatctgt
ctatttcgtt catccatagt tgcctgactc 4200cccgtcgtgt agataactac
gatacgggag ggcttaccat ctggccccag tgctgcaatg 4260ataccgcgag
acccacgctc accggctcca gatttatcag caataaacca gccagccgga
4320agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
tattaattgt 4380tgccgggaag ctagagtaag tagttcgcca gttaatagtt
tgcgcaacgt tgttgccatt 4440gctacaggca tcgtggtgtc acgctcgtcg
tttggtatgg cttcattcag ctccggttcc 4500caacgatcaa ggcgagttac
atgatccccc atgttgtgca aaaaagcggt tagctccttc 4560ggtcctccga
tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
4620gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
gactggtgag 4680tactcaacca agtcattctg agaatagtgt atgcggcgac
cgagttgctc ttgcccggcg 4740tcaatacggg ataataccgc gccacatagc
agaactttaa aagtgctcat cattggaaaa 4800cgttcttcgg ggcgaaaact
ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa 4860cccactcgtg
cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
4920gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
gaaatgttga 4980atactcatac tcttcctttt tcaatattat tgaagcattt
atcagggtta ttgtctcatg 5040agcggataca tatttgaatg tatttagaaa
aataaacaaa taggggttcc gcgcacattt 5100ccccgaaaag tgccacctga
cgtcgacgga tcgggagatc tcccgatccc ctatggtcga 5160ctctcagtac
aatctgctct gatgccgcat agttaagcca gtatctgctc cctgcttgtg
5220tgttggaggt cgctgagtag tgcgcgagca aaatttaagc tacaacaagg
caaggcttga 5280ccgacaattg catgaagaat ctgcttaggg ttaggcgttt
tgcgctgctt cgcgatgtac 5340gggccagata tacgcgttga cattgattat
tgactagtta ttaatagtaa tcaattacgg 5400ggtcattagt tcatagccca
tatatggagt tccgcgttac ataacttacg gtaaatggcc 5460cgcctggctg
accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca
5520tagtaacgcc aatagggact ttccattgac gtcaatgggt ggactattta
cggtaaactg 5580cccacttggc agtacatcaa gtgtatcata tgccaagtac
gccccctatt gacgtcaatg 5640acggtaaatg gcccgcctgg cattatgccc
agtacatgac cttatgggac tttcctactt 5700ggcagtacat ctacgtatta
gtcatcgcta ttaccatggt gatgcggttt tggcagtaca 5760tcaatgggcg
tggatagcgg tttgactcac ggggatttcc aagtctccac cccattgacg
5820tcaatgggag tttgttttgg caccaaaatc aacgggactt tccaaaatgt
cgtaacaact 5880ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg
ggaggtctat ataagcagag 5940ctctctggct aactagagaa cccactgctt
actggcttat cgaaat 59864612PRTHomo sapiens 46Lys Ile Val Ile Leu Leu
Asp Tyr Met Phe Gln Asp 1 5 10 4712PRTHomo sapiens 47Ala Ile Arg
Asp Tyr Ser Phe Lys Arg Leu Tyr Arg 1 5 10 4812PRTHomo sapiens
48Leu Lys Val Leu Glu Ile Ser His Trp Pro Tyr Leu 1 5 10
4915PRTHomo sapiens 49Asn Leu Thr Ala Val Pro Tyr Leu Ala Val Arg
His Leu Val Tyr 1 5 10 15 509PRTHomo sapiens 50Tyr Phe Thr Cys Arg
Arg Ala Arg Ile 1 5 5115PRTHomo sapiens 51Asp Val Leu Leu Pro Asn
Tyr Phe Thr Cys Arg Arg Ala Arg Ile 1 5 10 15
* * * * *
References