U.S. patent application number 17/478587 was filed with the patent office on 2022-06-09 for anti-trem2 antibodies and methods of use thereof.
The applicant listed for this patent is Denali Therapeutics Inc.. Invention is credited to Mark S. Dennis, Zhenyu Gu, Mihalis Kariolis, Cathal Mahon, Kathryn M. Monroe, Joshua I. Park, Rachel Prorok, Adam P. Silverman, Bettina Van Lengerich.
Application Number | 20220177576 17/478587 |
Document ID | / |
Family ID | 1000006156861 |
Filed Date | 2022-06-09 |
United States Patent
Application |
20220177576 |
Kind Code |
A1 |
Dennis; Mark S. ; et
al. |
June 9, 2022 |
ANTI-TREM2 ANTIBODIES AND METHODS OF USE THEREOF
Abstract
In one aspect, antibodies that specifically bind to a human
triggering receptor expressed on myeloid cells 2 (TREM2) protein
are provided. In some embodiments, the antibody decreases levels of
soluble TREM2 (sTREM2). In some embodiments, the antibody enhances
TREM2 activity.
Inventors: |
Dennis; Mark S.; (South San
Francisco, CA) ; Gu; Zhenyu; (South San Francisco,
CA) ; Kariolis; Mihalis; (South San Francisco,
CA) ; Mahon; Cathal; (South San Francisco, CA)
; Monroe; Kathryn M.; (South San Francisco, CA) ;
Park; Joshua I.; (South San Francisco, CA) ; Prorok;
Rachel; (South San Francisco, CA) ; Silverman; Adam
P.; (South San Francisco, CA) ; Van Lengerich;
Bettina; (South San Francisco, CA) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
Denali Therapeutics Inc. |
South San Francisco |
CA |
US |
|
|
Family ID: |
1000006156861 |
Appl. No.: |
17/478587 |
Filed: |
September 17, 2021 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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17151006 |
Jan 15, 2021 |
11124567 |
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17478587 |
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PCT/US2021/013200 |
Jan 13, 2021 |
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17151006 |
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62960663 |
Jan 13, 2020 |
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63070728 |
Aug 26, 2020 |
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63091717 |
Oct 14, 2020 |
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Current U.S.
Class: |
1/1 |
Current CPC
Class: |
C07K 2317/565 20130101;
A61K 2039/505 20130101; C07K 16/2803 20130101; C07K 2319/30
20130101 |
International
Class: |
C07K 16/28 20060101
C07K016/28 |
Claims
1-88. (canceled)
89. A method of treating a neurodegenerative disease in a subject,
comprising administering to the subject an isolated antibody that
specifically binds to a human triggering receptor expressed on
myeloid cells 2 (TREM2), wherein the antibody comprises: (a) a
variable region comprising: i. a CDR-H1 sequence comprising the
sequence of G-F-T-F-T-.alpha..sub.6-F-Y-M-S (SEQ ID NO:28), wherein
.alpha..sub.6 is D or N; ii. a CDR-H2 sequence comprising the
sequence of
V-I-R-N-.beta..sub.5-.beta..sub.6-N-.beta..sub.8-Y-T-.beta..sub.11-.beta.-
.sub.12-Y-N-P-S-V-K-G (SEQ ID NO:29), wherein .beta..sub.5 is K or
R; .beta..sub.6 is A or P; .beta..sub.8 is G or A; .beta..sub.11 is
A or T; and .beta..sub.12 is G or D; iii. a CDR-H3 sequence
comprising the sequence of
.gamma..sub.1-R-L-.gamma..sub.4-Y-G-F-D-Y (SEQ ID NO:30), wherein
.gamma..sub.1 is A or T; and .gamma..sub.4 is T or S; iv. a CDR-L1
sequence comprising the sequence of
Q-S-S-K-S-L-L-H-S-.delta..sub.10-G-K-T-Y-L-N (SEQ ID NO:31),
wherein .delta..sub.10 is N or T; v. a CDR-L2 sequence comprising
the sequence WMSTRAS (SEQ ID NO:8); and vi. a CDR-L3 sequence
comprising the sequence of Q-Q-F-L-E-.PHI..sub.6-P-F-T (SEQ ID
NO:32), wherein .PHI..sub.6 is Y or F: (b) a first Fc polypeptide
that specifically binds to human transferrin receptor 1 and
comprises a sequence having at least 90% sequence identity to SEQ
ID NO:49; and (c) a second Fc polypeptide.
90. The method of claim 89, wherein the neurodegenerative disease
is selected from the group consisting of: Alzheimer's disease,
primary age-related tauopathy, progressive supranuclear palsy
(PSP), frontotemporal dementia, frontotemporal dementia with
parkinsonism linked to chromosome 17, argyrophilic grain dementia,
amyotrophic lateral sclerosis, amyotrophic lateral
sclerosis/parkinsonism-dementia complex of Guam (ALS-PDC),
corticobasal degeneration, chronic traumatic encephalopathy,
Creutzfeldt-Jakob disease, dementia pugilistica, diffuse
neurofibrillary tangles with calcification, Down's syndrome,
familial British dementia, familial Danish dementia,
Gerstmann-Straussler-Scheinker disease, globular glial tauopathy,
Guadeloupean parkinsonism with dementia, Guadelopean PSP,
Hallevorden-Spatz disease, hereditary diffuse leukoencephalopathy
with spheroids (HDLS), Huntington's disease, inclusion-body
myositis, multiple system atrophy, myotonic dystrophy, Nasu-Hakola
disease, neurofibrillary tangle-predominant dementia, Niemann-Pick
disease type C, pallido-ponto-nigral degeneration, Parkinson's
disease, Pick's disease, postencephalitic parkinsonism, prion
protein cerebral amyloid angiopathy, progressive subcortical
gliosis, subacute sclerosing panencephalitis, and tangle only
dementia.
91. A method of decreasing levels of sTREM2 in a subject having a
neurodegenerative disease, comprising administering to the subject
an isolated antibody that specifically binds to a human triggering
receptor expressed on myeloid cells 2 (TREM2), wherein the antibody
comprises: (a) a variable region comprising: i. a CDR-H1 sequence
comprising the sequence of G-F-T-F-T-.alpha..sub.6-F-Y-M-S (SEQ ID
NO:28), wherein .alpha..sub.6 is D or N; ii. a CDR-H2 sequence
comprising the sequence of
V-I-R-N-.beta..sub.5-.beta..sub.6-N-.beta..sub.8-Y-T-.beta..sub.11-.beta.-
.sub.12-Y-N-P-S-V-K-G (SEQ ID NO:29), wherein .beta..sub.5 is K or
R; .beta..sub.6 is A or P; .beta..sub.8 is G or A; .beta..sub.11 is
A or T; and .beta..sub.12 is G or D; iii. a CDR-H3 sequence
comprising the sequence of
.gamma..sub.1-R-L-.gamma..sub.4-Y-G-F-D-Y (SEQ ID NO:30), wherein
.gamma..sub.1 is A or T; and .gamma..sub.4 is T or S; iv. a CDR-L1
sequence comprising the sequence of
Q-S-S-K-S-L-L-H-S-.delta..sub.10-G-K-T-Y-L-N (SEQ ID NO:31),
wherein .delta..sub.10 is N or T; v. a CDR-L2 sequence comprising
the sequence WMSTRAS (SEQ ID NO:8); and vi. a CDR-L3 sequence
comprising the sequence of Q-Q-F-L-E-.PHI..sub.6-P-F-T (SEQ ID
NO:32), wherein .PHI..sub.6 is Y or F: (b) a first Fc polypeptide
that specifically binds to human transferrin receptor 1 and
comprises a sequence having at least 90% sequence identity to SEQ
ID NO:49; and (c) a second Fc polypeptide.
92. A method of enhancing TREM2 activity in a subject having a
neurodegenerative disease, comprising administering to the subject
an isolated antibody that specifically binds to a human triggering
receptor expressed on myeloid cells 2 (TREM2), wherein the antibody
comprises: (a) a variable region comprising: i. a CDR-H1 sequence
comprising the sequence of G-F-T-F-T-.alpha..sub.6-F-Y-M-S (SEQ ID
NO:28), wherein .alpha..sub.6 is D or N; ii. a CDR-H2 sequence
comprising the sequence of
V-I-R-N-.beta..sub.5-.beta..sub.6-N-.beta..sub.8-Y-T-.beta..sub.11-.beta.-
.sub.12-Y-N-P-S-V-K-G (SEQ ID NO:29), wherein .beta..sub.5 is K or
R; .beta..sub.6 is A or P; .beta..sub.8 is G or A; .beta..sub.11 is
A or T; and .beta..sub.12 is G or D; iii. a CDR-H3 sequence
comprising the sequence of
.gamma..sub.1-R-L-.gamma..sub.4-Y-G-F-D-Y (SEQ ID NO:30), wherein
.gamma..sub.1 is A or T; and .gamma..sub.4 is T or S; iv. a CDR-L1
sequence comprising the sequence of
Q-S-S-K-S-L-L-H-S-.delta..sub.10-G-K-T-Y-L-N (SEQ ID NO:31),
wherein .delta..sub.10 is N or T; v. a CDR-L2 sequence comprising
the sequence WMSTRAS (SEQ ID NO:8); and vi. a CDR-L3 sequence
comprising the sequence of Q-Q-F-L-E-.PHI..sub.6-P-F-T (SEQ ID
NO:32), wherein .PHI..sub.6 is Y or F; (b) a first Fc polypeptide
that specifically binds to human transferrin receptor 1 and
comprises a sequence having at least 90% sequence identity to SEQ
ID NO:49; and (c) a second Fc polypeptide.
93-102. (canceled)
103. The method of claim 89, wherein: i. the CDR-H1 sequence is
selected from the group consisting of SEQ ID NOS:4 and 12; ii. the
CDR-H2 sequence is selected from the group consisting of SEQ ID
NOS:5, 13, and 25; iii. the CDR-H3 sequence is selected from the
group consisting of SEQ ID NOS:6, 14, and 17; iv. the CDR-L1
sequence is selected from the group consisting of SEQ ID NOS:7 and
23; v. the CDR-L2 sequence comprises the sequence WMSTRAS (SEQ ID
NO:8); and vi. the CDR-L3 sequence is selected from the group
consisting of SEQ ID NOS:9 and 18.
104. The method of claim 89, wherein the variable region comprises
a CDR-H1 comprising the amino acid sequence of SEQ ID NO:4, a
CDR-H2 comprising the amino acid sequence of SEQ ID NO:25, a CDR-H3
comprising the amino acid sequence of SEQ ID NO:17, a CDR-L1
comprising the amino acid sequence of SEQ ID NO:23, a CDR-L2
comprising the amino acid sequence of SEQ ID NO:8, and a CDR-L3
comprising the amino acid sequence of SEQ ID NO:18.
105. The method of claim 89, wherein the variable region comprises
a V.sub.H sequence that has at least 85% sequence identity to SEQ
ID NO:24 and a V.sub.L sequence has at least 85% sequence identity
to SEQ ID NO:22.
106. The method of claim 89, wherein the first Fc polypeptide
comprises: Trp, Leu, or Glu at position 380; Tyr or Phe at position
384; Thr at position 386; Glu at position 387; Trp at position 388;
Ser, Ala, or Val at position 389; Ser or Asn at position 390; Thr
or Ser at position 413; Glu or Ser at position 415; Glu at position
416; and Phe at position 421, according to EU numbering.
107. The method of claim 106, wherein: (a) the first Fc polypeptide
has a T366W substitution and the second Fc polypeptide has T366S,
L368A, and Y407V substitutions, according to EU numbering; (b) the
first Fc polypeptide and/or the second Fc polypeptide comprises
amino acid substitutions of Ala at position 234 and Ala at position
235, according to EU numbering; and/or (c) the first Fc polypeptide
and/or the second Fc polypeptide comprises amino acid substitutions
of Leu at position 428 and Ser at position 434, according to EU
numbering.
108. The method of claim 89, wherein the first Fc polypeptide
comprises the sequence of SEQ ID NO:41 or 64, and the second Fc
polypeptide comprises the sequence of SEQ ID NO:39 or 63.
109. The method of claim 108, comprising: (i) a first heavy chain
(HC) comprising a V.sub.H comprising SEQ ID NO:24 and the first Fc
polypeptide comprising SEQ ID NO:41 or 64; (ii) a second heavy
chain (HC) comprising a V.sub.H comprising SEQ ID NO:24 and the
second Fc polypeptide comprising SEQ ID NO:39 or 63; and (iii) two
light chains each comprising a V.sub.L comprising SEQ ID NO:22.
110. The method of claim 109, comprising: (i) a first heavy chain
(HC) that comprises or consists of the amino acid sequence set
forth in SEQ ID NO:42 or 65; (ii) a second HC that comprises or
consists of the amino acid sequence set forth in SEQ ID NO:53 or
73; and (iii) a first and a second light chain (LC) that each
comprises or consists of the amino acid sequence set forth in SEQ
ID NO:54.
111. The method of claim 89, wherein the first Fc polypeptide
comprises the sequence of SEQ ID NO:44 or 66, and the second Fc
polypeptide comprises the sequence of SEQ ID NO:39 or 63.
112. The method of claim 111, comprising: (i) a first heavy chain
(HC) comprising a V.sub.H comprising SEQ ID NO:24 and the first Fc
polypeptide comprising SEQ ID NO:44 or 66; (ii) a second heavy
chain (HC) comprising a V.sub.H comprising SEQ ID NO:24 and the
second Fc polypeptide comprising SEQ ID NO:39 or 63; and (iii) two
light chains each comprising a V.sub.L comprising SEQ ID NO:22.
113. The method of claim 112, comprising: (i) a first heavy chain
(HC) that comprises or consists of the amino acid sequence set
forth in SEQ ID NO:45 or 67; (ii) a second HC that comprises or
consists of the amino acid sequence set forth in SEQ ID NO:53 or
73; and (iii) a first and a second light chain (LC) that each
comprises or consists of the amino acid sequence set forth in SEQ
ID NO:54.
114. The method of claim 89, wherein the first Fc polypeptide
comprises the sequence of SEQ ID NO:47 or 68, and the second Fc
polypeptide comprises the sequence of SEQ ID NO:39 or 63.
115. The method of claim 114, comprising: (i) a first heavy chain
(HC) comprising a V.sub.H comprising SEQ ID NO:24 and the first Fc
polypeptide comprising SEQ ID NO:47 or 68; (ii) a second heavy
chain (HC) comprising a V.sub.H comprising SEQ ID NO:24 and the
second Fc polypeptide comprising SEQ ID NO:39 or 63; and (iii) two
light chains each comprising a V.sub.L comprising SEQ ID NO:22.
116. The method of claim 115, comprising: (i) a first heavy chain
(HC) that comprises or consists of the amino acid sequence set
forth in SEQ ID NO:48 or 69; (ii) a second HC that comprises or
consists of the amino acid sequence set forth in SEQ ID NO:53 or
73; and (iii) a first and a second light chain (LC) that each
comprises or consists of the amino acid sequence set forth in SEQ
ID NO:54.
117. The method of claim 89, wherein the first Fc polypeptide
comprises the sequence of SEQ ID NO:47 or 68, and the second Fc
polypeptide comprises the sequence of SEQ ID NO:61 or 84.
118. The method of claim 117, comprising: (i) a first heavy chain
(HC) comprising a V.sub.H comprising SEQ ID NO:24 and the first Fc
polypeptide comprising SEQ ID NO:47 or 68; (ii) a second heavy
chain (HC) comprising a V.sub.H comprising SEQ ID NO:24 and the
second Fc polypeptide comprising SEQ ID NO:61 or 84; and (iii) two
light chains each comprising a V.sub.L comprising SEQ ID NO:22.
119. The method of claim 118, comprising: (i) a first heavy chain
(HC) that comprises or consists of the amino acid sequence set
forth in SEQ ID NO:48 or 69; (ii) a second HC that comprises or
consists of the amino acid sequence set forth in SEQ ID NO:52 or
72; and (iii) a first and a second light chain (LC) that each
comprises or consists of the amino acid sequence set forth in SEQ
ID NO:54.
120. The method of claim 89, wherein the first Fc polypeptide
comprises the sequence of SEQ ID NO:50 or 70, and the second Fc
polypeptide comprises the sequence of SEQ ID NO:39 or 63.
121. The method of claim 120, comprising: (i) a first heavy chain
(HC) comprising a V.sub.H comprising SEQ ID NO:24 and the first Fc
polypeptide comprising SEQ ID NO:50 or 70; (ii) a second heavy
chain (HC) comprising a V.sub.H comprising SEQ ID NO:24 and the
second Fc polypeptide comprising SEQ ID NO:39 or 63; and (iii) two
light chains each comprising a V.sub.L comprising SEQ ID NO:22.
122. The method of claim 121, comprising: (i) a first heavy chain
(HC) that comprises or consists of the amino acid sequence set
forth in SEQ ID NO:51 or 71; (ii) a second HC that comprises or
consists of the amino acid sequence set forth in SEQ ID NO:53 or
73; and (iii) a first and a second light chain (LC) that each
comprises or consists of the amino acid sequence set forth in SEQ
ID NO:54.
123. A method of treating a neurodegenerative disease in a subject,
comprising administering to the subject an isolated antibody that
specifically binds to a human triggering receptor expressed on
myeloid cells 2 (TREM2), wherein the antibody comprises: (i) a
first heavy chain (HC) comprising a V.sub.H comprising SEQ ID NO:24
and a first Fc polypeptide comprising SEQ ID NO:47; (ii) a second
heavy chain (HC) comprising a V.sub.H comprising SEQ ID NO:24 and a
second Fc polypeptide comprising SEQ ID NO:39; and (iii) two light
chains each comprising a V.sub.L comprising SEQ ID NO:22.
124. The method of claim 123, comprising: (i) a first heavy chain
(HC) that consists of the amino acid sequence set forth in SEQ ID
NO:48; (ii) a second HC that consists of the amino acid sequence
set forth in SEQ ID NO:53; and (iii) a first and a second light
chain (LC) that each consists of the amino acid sequence set forth
in SEQ ID NO:54.
125. A method of treating a neurodegenerative disease in a subject,
comprising administering to the subject an isolated antibody that
specifically binds to a human triggering receptor expressed on
myeloid cells 2 (TREM2), wherein the antibody comprises: (i) a
first heavy chain (HC) comprising a V.sub.H comprising SEQ ID NO:24
and a first Fc polypeptide comprising SEQ ID NO:68; (ii) a second
heavy chain (HC) comprising a V.sub.H comprising SEQ ID NO:24 and a
second Fc polypeptide comprising SEQ ID NO:63; and (iii) two light
chains each comprising a V.sub.L comprising SEQ ID NO:22.
126. The method of claim 125, comprising: (i) a first heavy chain
(HC) that consists of the amino acid sequence set forth in SEQ ID
NO:69; (ii) a second HC that consists of the amino acid sequence
set forth in SEQ ID NO:73; and (iii) a first and a second light
chain (LC) that each consists of the amino acid sequence set forth
in SEQ ID NO:54.
127. A method of treating a neurodegenerative disease in a subject,
comprising administering to the subject an isolated antibody that
specifically binds to a human triggering receptor expressed on
myeloid cells 2 (TREM2), wherein the antibody comprises: (a) a
variable region comprising: i. a CDR-H1 sequence comprising the
sequence of G-F-T-F-T-.alpha..sub.6-F-Y-M-S (SEQ ID NO:28), wherein
.alpha..sub.6 is D or N; ii. a CDR-H2 sequence comprising the
sequence of
V-I-R-N-.beta..sub.5-.beta..sub.6-N-.beta..sub.8-Y-T-.beta..sub.11-.beta.-
.sub.12-Y-N-P-S-V-K-G (SEQ ID NO:29), wherein .beta..sub.5 is K or
R; .beta..sub.6 is A or P; .beta..sub.8 is G or A; .DELTA..sub.11
is A or T; and .beta..sub.12 is G or D; iii. a CDR-H3 sequence
comprising the sequence of
.gamma..sub.1-R-L-.gamma..sub.4-Y-G-F-D-Y (SEQ ID NO:30), wherein
.gamma..sub.1 is A or T; and .gamma..sub.4 is T or S; iv. a CDR-L1
sequence comprising the sequence of
Q-S-S-K-S-L-L-H-S-.delta..sub.10-G-K-T-Y-L-N (SEQ ID NO:31),
wherein .delta..sub.10 is N or T; v. a CDR-L2 sequence comprising
the sequence WMSTRAS (SEQ ID NO:8); and vi. a CDR-L3 sequence
comprising the sequence of Q-Q-F-L-E-.PHI..sub.6-P-F-T (SEQ ID
NO:32), wherein .PHI..sub.6 is Y or F; (b) a first Fc polypeptide
that specifically binds to human transferrin receptor 1 and
comprises: Trp, Leu, or Glu at position 380; Tyr or Phe at position
384; Thr at position 386; Glu at position 387; Trp at position 388;
Ser, Ala, or Val at position 389; Ser or Asn at position 390; Thr
or Ser at position 413; Glu or Ser at position 415; Glu at position
416; and Phe at position 421, according to EU numbering; and (c) a
second Fc polypeptide.
128. The method of claim 127, wherein: (a) the first Fc polypeptide
has a T366W substitution and the second Fc polypeptide has T366S,
L368A, and Y407V substitutions, according to EU numbering; (b) the
first Fc polypeptide and/or the second Fc polypeptide comprises
amino acid substitutions of Ala at position 234 and Ala at position
235, according to EU numbering; and/or (c) the first Fc polypeptide
and/or the second Fc polypeptide comprises amino acid substitutions
of Leu at position 428 and Ser at position 434, according to EU
numbering.
129. The method of claim 89, wherein the neurodegenerative disease
is Alzheimer's disease.
130. A method of treating Alzheimer's disease in a subject,
comprising administering to the subject an isolated antibody that
specifically binds to a human triggering receptor expressed on
myeloid cells 2 (TREM2), wherein the antibody comprises: (i) a
first heavy chain (HC) comprising a V.sub.H comprising SEQ ID NO:24
and a first Fc polypeptide comprising SEQ ID NO:47; (ii) a second
heavy chain (HC) comprising a V.sub.H comprising SEQ ID NO:24 and a
second Fc polypeptide comprising SEQ ID NO:39; and (iii) two light
chains each comprising a V.sub.L comprising SEQ ID NO:22.
131. The method of claim 130, comprising: (i) a first heavy chain
(HC) that consists of the amino acid sequence set forth in SEQ ID
NO:48; (ii) a second HC that consists of the amino acid sequence
set forth in SEQ ID NO:53; and (iii) a first and a second light
chain (LC) that each consists of the amino acid sequence set forth
in SEQ ID NO:54.
Description
CROSS-REFERENCE TO RELATED APPLICATIONS
[0001] The present application is a continuation of U.S.
application Ser. No. 17/151,006, filed Jan. 15, 2021, which is a
continuation of International Patent Application No.
PCT/US2021/013200, filed on Jan. 13, 2021, which claims priority to
U.S. Provisional Patent Application No. 62/960,663, filed on Jan.
13, 2020, U.S. Provisional Patent Application No. 63/070,728, filed
on Aug. 26, 2020, and U.S. Provisional Patent Application No.
63/091,717, filed on Oct. 14, 2020, the disclosures of which are
incorporated herein by reference in their entirety for all
purposes.
REFERENCE TO A SEQUENCE LISTING
[0002] The Sequence Listing written in file
102342-004040US-1268163_SL.txt created on Sep. 2, 2021, 160,537
bytes, machine format IBM-PC, MS-Windows operating system, is
hereby incorporated by reference in its entirety for all
purposes.
BACKGROUND
[0003] Triggering receptor expressed on myeloid cells-2 (TREM2) is
a transmembrane receptor that is expressed on microglia and is
believed to function in regulating phagocytosis, cell survival, and
the production of pro-inflammatory cytokines. Mutations in TREM2
have been identified in neurodegenerative diseases including
Alzheimer's disease, Nasu-Hakola disease, Parkinson's disease,
amyotrophic lateral sclerosis, and frontotemporal dementia.
Additionally, altered levels of soluble TREM2 (sTREM2) have been
reported in the cerebrospinal fluid of patients having Alzheimer's
disease or frontotemporal dementia who have a mutation in
TREM2.
[0004] There remains a need for therapeutic agents that modulate
TREM2 activity or levels of sTREM2.
BRIEF SUMMARY
[0005] In one aspect, antibodies that specifically binds to a human
triggering receptor expressed on myeloid cells 2 (TREM2) are
provided. In some embodiments, the antibodies comprise a modified
Fc polypeptide that can bind to a transferrin receptor protein. In
any of the herein disclosed embodiments, the antibody can comprise
CDRs, a V.sub.H, and/or a V.sub.L according to any of the exemplary
sequences provided herein, and can further comprise a modified Fc
polypeptide comprising transferrin receptor-binding mutations as
set forth herein.
[0006] In some embodiments, an antibody that specifically binds to
TREM2 comprises:
[0007] (a) a variable region comprising: [0008] i. a CDR-H1
sequence comprising the sequence of
G-F-T-F-T-.alpha..sub.6-F-Y-M-S(SEQ ID NO:28), wherein
.alpha..sub.6 is D or N; [0009] ii. a CDR-H2 sequence comprising
the sequence of
V-I-R-N-.beta..sub.5-.beta..sub.6-N-.beta..sub.8-Y-T-.beta..sub.11-.beta.-
.sub.12-Y-N-P-S-V-K-G (SEQ ID NO:29), wherein .beta..sub.5 is K or
R; .beta..sub.6 is A or P; .beta..sub.8 is G or A; .beta..sub.11 is
A or T; and .beta..sub.12 is G or D; [0010] iii. a CDR-H3 sequence
comprising the sequence of
.gamma..sub.1-R-L-.gamma..sub.4-Y-G-F-D-Y (SEQ ID NO:30), wherein
.gamma..sub.1 is A or T; and .gamma..sub.4 is T or S; [0011] iv. a
CDR-L1 sequence comprising the sequence of
Q-S-S-K-S-L-L-H-S-.delta..sub.10-G-K-T-Y-L-N (SEQ ID NO:31),
wherein .delta..sub.10 is N or T; [0012] v. a CDR-L2 sequence
comprising the sequence WMSTRAS (SEQ ID NO:8); and [0013] vi. a
CDR-L3 sequence comprising the sequence of
Q-Q-F-L-E-.PHI..sub.6-P-F-T (SEQ ID NO:32), wherein .PHI..sub.6 is
Y or F;
[0014] (b) a first Fc polypeptide that is modified to specifically
bind to a transferrin receptor; and
[0015] (c) a second Fc polypeptide.
[0016] In some embodiments, the first Fc polypeptide and the second
Fc polypeptide associate to form an Fc dimer.
[0017] In some embodiments, the CDR-H1 sequence is selected from
SEQ ID NOS:4 or 12. In some embodiments, the CDR-H2 sequence is
selected from SEQ ID NOS:5, 13, or 25. In some embodiments, the
CDR-H3 sequence is selected from SEQ ID NOS:6, 14, or 17. In some
embodiments, the CDR-L1 sequence is selected from SEQ ID NOS:7 or
23. In some embodiments, the CDR-L3 sequence is selected from SEQ
ID NOS:9 or 18.
[0018] In some embodiments, the variable region comprises:
[0019] (a) a CDR-H1 comprising the amino acid sequence of SEQ ID
NO:4, a CDR-H2 comprising the amino acid sequence of SEQ ID NO:5, a
CDR-H3 comprising the amino acid sequence of SEQ ID NO:17, a CDR-L1
comprising the amino acid sequence of SEQ ID NO:7, a CDR-L2
comprising the amino acid sequence of SEQ ID NO:8, and a CDR-L3
comprising the amino acid sequence of SEQ ID NO:18; or
[0020] (b) a CDR-H1 comprising the amino acid sequence of SEQ ID
NO:4, a CDR-H2 comprising the amino acid sequence of SEQ ID NO:5, a
CDR-H3 comprising the amino acid sequence of SEQ ID NO:17, a CDR-L1
comprising the amino acid sequence of SEQ ID NO:23, a CDR-L2
comprising the amino acid sequence of SEQ ID NO:8, and a CDR-L3
comprising the amino acid sequence of SEQ ID NO:18; or
[0021] (c) a CDR-H1 comprising the amino acid sequence of SEQ ID
NO:4, a CDR-H2 comprising the amino acid sequence of SEQ ID NO:25,
a CDR-H3 comprising the amino acid sequence of SEQ ID NO:17, a
CDR-L1 comprising the amino acid sequence of SEQ ID NO:7, a CDR-L2
comprising the amino acid sequence of SEQ ID NO:8, and a CDR-L3
comprising the amino acid sequence of SEQ ID NO:18; or
[0022] (d) a CDR-H1 comprising the amino acid sequence of SEQ ID
NO:4, a CDR-H2 comprising the amino acid sequence of SEQ ID NO:25,
a CDR-H3 comprising the amino acid sequence of SEQ ID NO:17, a
CDR-L1 comprising the amino acid sequence of SEQ ID NO:23, a CDR-L2
comprising the amino acid sequence of SEQ ID NO:8, and a CDR-L3
comprising the amino acid sequence of SEQ ID NO:18; or
[0023] (e) a CDR-H1 comprising the amino acid sequence of SEQ ID
NO:4, a CDR-H2 comprising the amino acid sequence of SEQ ID NO:5, a
CDR-H3 comprising the amino acid sequence of SEQ ID NO:6, a CDR-L1
comprising the amino acid sequence of SEQ ID NO:7, a CDR-L2
comprising the amino acid sequence of SEQ ID NO:8, and a CDR-L3
comprising the amino acid sequence of SEQ ID NO:9; or
[0024] (f) a CDR-H1 comprising the amino acid sequence of SEQ ID
NO:12, a CDR-H2 comprising the amino acid sequence of SEQ ID NO:13,
a CDR-H3 comprising the amino acid sequence of SEQ ID NO:14, a
CDR-L1 comprising the amino acid sequence of SEQ ID NO:7, a CDR-L2
comprising the amino acid sequence of SEQ ID NO:8, and a CDR-L3
comprising the amino acid sequence of SEQ ID NO:9; or
[0025] (g) a CDR-H1 comprising the amino acid sequence of SEQ ID
NO:4, a CDR-H2 comprising the amino acid sequence of SEQ ID NO:25,
a CDR-H3 comprising the amino acid sequence of SEQ ID NO:17, a
CDR-L1 comprising the amino acid sequence of SEQ ID NO:7, a CDR-L2
comprising the amino acid sequence of SEQ ID NO:8, and a CDR-L3
comprising the amino acid sequence of SEQ ID NO:9.
[0026] In some embodiments, the variable region comprises a V.sub.H
sequence that has at least 85% sequence identity to any one of SEQ
ID NOS:2, 10, 15, 19, 21, 24, and 26.
[0027] In certain embodiments, the V.sub.H sequence has at least
90% sequence identity to SEQ ID NO: 15. In certain embodiments, the
V.sub.H sequence has at least 95% sequence identity to SEQ ID NO:
15. In certain embodiments, the V.sub.H sequence comprises SEQ ID
NO:15.
[0028] In certain embodiments, the V.sub.H sequence has at least
90% sequence identity to SEQ ID NO:24. In certain embodiments, the
V.sub.H sequence has at least 95% sequence identity to SEQ ID
NO:24. In certain embodiments, the V.sub.H sequence comprises SEQ
ID NO:24.
[0029] In some embodiments of this aspect, the variable region
comprises a V.sub.L sequence has at least 85% sequence identity to
any one of SEQ ID NOS:3, 11, 16, 20, 22, and 27.
[0030] In certain embodiments, the V.sub.L sequence has at least
90% sequence identity to SEQ ID NO: 16. In certain embodiments, the
V.sub.L sequence has at least 95% sequence identity to SEQ ID NO:
16. In certain embodiments, the V.sub.L sequence comprises SEQ ID
NO:16.
[0031] In certain embodiments, the V.sub.L sequence has at least
90% sequence identity to SEQ ID NO:22. In certain embodiments, the
V.sub.L sequence has at least 95% sequence identity to SEQ ID
NO:22. In certain embodiments, the V.sub.L sequence comprises SEQ
ID NO:22.
[0032] In certain embodiments, the V.sub.L sequence has at least
90% sequence identity to SEQ ID NO:27. In certain embodiments, the
V.sub.L sequence has at least 95% sequence identity to SEQ ID
NO:27. In certain embodiments, the V.sub.L sequence comprises SEQ
ID NO:27.
[0033] In some embodiments of this aspect, the variable region
comprises:
[0034] (a) a V.sub.H sequence comprising SEQ ID NO:15 and a V.sub.L
sequence comprising SEQ ID NO:16; or
[0035] (b) a V.sub.H sequence comprising SEQ ID NO:19 and a V.sub.L
sequence comprising SEQ ID NO:20; or
[0036] (c) a V.sub.H sequence comprising SEQ ID NO:21 and a V.sub.L
sequence comprising SEQ ID NO:20; or
[0037] (d) a V.sub.H sequence comprising SEQ ID NO:19 and a V.sub.L
sequence comprising SEQ ID NO:22; or
[0038] (e) a V.sub.H sequence comprising SEQ ID NO:21 and a V.sub.L
sequence comprising SEQ ID NO:22; or
[0039] (f) a V.sub.H sequence comprising SEQ ID NO:24 and a V.sub.L
sequence comprising SEQ ID NO:20; or
[0040] (g) a V.sub.H sequence comprising SEQ ID NO:26 and a V.sub.L
sequence comprising SEQ ID NO:20; or
[0041] (h) a V.sub.H sequence comprising SEQ ID NO:24 and a V.sub.L
sequence comprising SEQ ID NO:22; or
[0042] (i) a V.sub.H sequence comprising SEQ ID NO:26 and a V.sub.L
sequence comprising SEQ ID NO:22; or
[0043] (j) a V.sub.H sequence comprising SEQ ID NO:2 and a V.sub.L
sequence comprising SEQ ID NO:3; or
[0044] (k) a V.sub.H sequence comprising SEQ ID NO:10 and a V.sub.L
sequence comprising SEQ ID NO:11; or
[0045] (l) a V.sub.H sequence comprising SEQ ID NO:24 and a V.sub.L
sequence comprising SEQ ID NO:27.
[0046] In some embodiments of this aspect, the first Fc polypeptide
comprises: Trp, Leu, or Glu at position 380; Tyr or Phe at position
384; Thr at position 386; Glu at position 387; Trp at position 388;
Ser, Ala, or Val at position 389; Ser or Asn at position 390; Thr
or Ser at position 413; Glu or Ser at position 415; Glu at position
416; and Phe at position 421, according to EU numbering. In some
embodiments, the first Fc polypeptide binds to the apical domain of
the transferrin receptor. In particular embodiments, the antibody
has improved brain uptake compared to an antibody having a
wild-type Fc dimer.
[0047] In certain embodiments, the first Fc polypeptide has a T366W
substitution and the second Fc polypeptide has T366S, L368A, and
Y407V substitutions, according to EU numbering.
[0048] In other embodiments, the first Fc polypeptide has T366S,
L368A, and Y407V substitutions and the second Fc polypeptide has a
T366W substitution, according to EU numbering.
[0049] In some embodiments, the first Fc polypeptide and/or the
second Fc polypeptide comprises a modification that reduces
effector function. In certain embodiments, the modification that
reduces effector function comprises the substitutions of Ala at
position 234 and Ala at position 235, according to EU
numbering.
[0050] In some embodiments, the first Fc polypeptide and/or the
second Fc polypeptide comprises amino acid modifications relative
to the native Fc sequence that extend serum half-life. In certain
embodiments, the amino acid modifications comprise substitutions of
Leu at position 428 and Ser at position 434, according to EU
numbering.
[0051] In some embodiments of this aspect, the first Fc polypeptide
comprises the sequence of SEQ ID NO:41 or SEQ ID NO:64, and the
second Fc polypeptide comprises the sequence of SEQ ID NO:39 or SEQ
ID NO:63. In certain embodiments, the antibody comprises: (i) a
first heavy chain (HC) comprising a V.sub.H comprising SEQ ID NO:24
and the first Fc polypeptide comprising SEQ ID NO:41; (ii) a second
heavy chain (HC) comprising a V.sub.H comprising SEQ ID NO:24 and
the second Fc polypeptide comprising SEQ ID NO:39; and (iii) two
light chains each comprising a V.sub.L comprising SEQ ID NO:22. In
certain embodiments, the antibody comprises: (i) a first heavy
chain (HC) comprising a V.sub.H comprising SEQ ID NO:24 and the
first Fc polypeptide comprising SEQ ID NO:64; (ii) a second heavy
chain (HC) comprising a V.sub.H comprising SEQ ID NO:24 and the
second Fc polypeptide comprising SEQ ID NO:63; and (iii) two light
chains each comprising a V.sub.L comprising SEQ ID NO:22. In
particular embodiments, the antibody comprises: (i) a first heavy
chain (HC) that comprises or consists of the amino acid sequence
set forth in SEQ ID NO:42; (ii) a second HC that comprises or
consists of the amino acid sequence set forth in SEQ ID NO:53; and
(iii) a first and a second light chain (LC) that each comprises or
consists of the amino acid sequence set forth in SEQ ID NO:54. In
certain embodiments, the antibody comprises: (i) a first heavy
chain (HC) that comprises or consists of the amino acid sequence
set forth in SEQ ID NO:65; (ii) a second HC that comprises or
consists of the amino acid sequence set forth in SEQ ID NO:73; and
(iii) a first and a second light chain (LC) that each comprises or
consists of the amino acid sequence set forth in SEQ ID NO:54.
[0052] In some embodiments of this aspect, the first Fc polypeptide
comprises the sequence of SEQ ID NO:44 or SEQ ID NO:66, and the
second Fc polypeptide comprises the sequence of SEQ ID NO:39 or SEQ
ID NO:63. In certain embodiments, the antibody comprises: (i) a
first heavy chain (HC) comprising a V.sub.H comprising SEQ ID NO:24
and the first Fc polypeptide comprising SEQ ID NO:44; (ii) a second
heavy chain (HC) comprising a V.sub.H comprising SEQ ID NO:24 and
the second Fc polypeptide comprising SEQ ID NO:39; and (iii) two
light chains each comprising a V.sub.L comprising SEQ ID NO:22. In
certain embodiments, the antibody comprises: (i) a first heavy
chain (HC) comprising a V.sub.H comprising SEQ ID NO:24 and the
first Fc polypeptide comprising SEQ ID NO:66; (ii) a second heavy
chain (HC) comprising a V.sub.H comprising SEQ ID NO:24 and the
second Fc polypeptide comprising SEQ ID NO:63; and (iii) two light
chains each comprising a V.sub.L comprising SEQ ID NO:22. In
particular embodiments, the antibody comprises: (i) a first heavy
chain (HC) that comprises or consists of the amino acid sequence
set forth in SEQ ID NO:45; (ii) a second HC that comprises or
consists of the amino acid sequence set forth in SEQ ID NO:53; and
(iii) a first and a second light chain (LC) that each comprises or
consists of the amino acid sequence set forth in SEQ ID NO:54. In
certain embodiments, the antibody comprises: (i) a first heavy
chain (HC) that comprises or consists of the amino acid sequence
set forth in SEQ ID NO:67; (ii) a second HC that comprises or
consists of the amino acid sequence set forth in SEQ ID NO:73; and
(iii) a first and a second light chain (LC) that each comprises or
consists of the amino acid sequence set forth in SEQ ID NO:54.
[0053] In some embodiments of this aspect, the first Fc polypeptide
comprises the sequence of SEQ ID NO:47 or SEQ ID NO:68, and the
second Fc polypeptide comprises the sequence of SEQ ID NO:39 or SEQ
ID NO:63. In certain embodiments, the antibody comprises: (i) a
first heavy chain (HC) comprising a V.sub.H comprising SEQ ID NO:24
and the first Fc polypeptide comprising SEQ ID NO:47; (ii) a second
heavy chain (HC) comprising a V.sub.H comprising SEQ ID NO:24 and
the second Fc polypeptide comprising SEQ ID NO:39; and (iii) two
light chains each comprising a V.sub.L comprising SEQ ID NO:22. In
certain embodiments, the antibody comprises: (i) a first heavy
chain (HC) comprising a V.sub.H comprising SEQ ID NO:24 and the
first Fc polypeptide comprising SEQ ID NO:68; (ii) a second heavy
chain (HC) comprising a V.sub.H comprising SEQ ID NO:24 and the
second Fc polypeptide comprising SEQ ID NO:63; and (iii) two light
chains each comprising a V.sub.L comprising SEQ ID NO:22. In
particular embodiments, the antibody comprises: (i) a first heavy
chain (HC) that comprises or consists of the amino acid sequence
set forth in SEQ ID NO:48; (ii) a second HC that comprises or
consists of the amino acid sequence set forth in SEQ ID NO:53; and
(iii) a first and a second light chain (LC) that each comprises or
consists of the amino acid sequence set forth in SEQ ID NO:54. In
certain embodiments, the antibody comprises: (i) a first heavy
chain (HC) that comprises or consists of the amino acid sequence
set forth in SEQ ID NO:69; (ii) a second HC that comprises or
consists of the amino acid sequence set forth in SEQ ID NO:73; and
(iii) a first and a second light chain (LC) that each comprises or
consists of the amino acid sequence set forth in SEQ ID NO:54.
[0054] In some embodiments of this aspect, the first Fc polypeptide
comprises the sequence of SEQ ID NO:47 or SEQ ID NO:68, and the
second Fc polypeptide comprises the sequence of SEQ ID NO:61 or SEQ
ID NO:84. In certain embodiments, the antibody comprises: (i) a
first heavy chain (HC) comprising a V.sub.H comprising SEQ ID NO:24
and the first Fc polypeptide comprising SEQ ID NO:47; (ii) a second
heavy chain (HC) comprising a V.sub.H comprising SEQ ID NO:24 and
the second Fc polypeptide comprising SEQ ID NO:61; and (iii) two
light chains each comprising a V.sub.L comprising SEQ ID NO:22. In
certain embodiments, the antibody comprises: (i) a first heavy
chain (HC) comprising a V.sub.H comprising SEQ ID NO:24 and the
first Fc polypeptide comprising SEQ ID NO:68; (ii) a second heavy
chain (HC) comprising a V.sub.H comprising SEQ ID NO:24 and the
second Fc polypeptide comprising SEQ ID NO:84; and (iii) two light
chains each comprising a V.sub.L comprising SEQ ID NO:22. In
particular embodiments, the antibody comprises: (i) a first heavy
chain (HC) that comprises or consists of the amino acid sequence
set forth in SEQ ID NO:48; (ii) a second HC that comprises or
consists of the amino acid sequence set forth in SEQ ID NO:52; and
(iii) a first and a second light chain (LC) that each comprises or
consists of the amino acid sequence set forth in SEQ ID NO:54. In
certain embodiments, the antibody comprises: (i) a first heavy
chain (HC) that comprises or consists of the amino acid sequence
set forth in SEQ ID NO:69; (ii) a second HC that comprises or
consists of the amino acid sequence set forth in SEQ ID NO:72; and
(iii) a first and a second light chain (LC) that each comprises or
consists of the amino acid sequence set forth in SEQ ID NO:54.
[0055] In some embodiments of this aspect, the first Fc polypeptide
comprises the sequence of SEQ ID NO:50 or SEQ ID NO:70, and the
second Fc polypeptide comprises the sequence of SEQ ID NO:39 or SEQ
ID NO:63. In certain embodiments, the antibody comprises: (i) a
first heavy chain (HC) comprising a V.sub.H comprising SEQ ID NO:24
and the first Fc polypeptide comprising SEQ ID NO:50; (ii) a second
heavy chain (HC) comprising a V.sub.H comprising SEQ ID NO:24 and
the second Fc polypeptide comprising SEQ ID NO:39; and (iii) two
light chains each comprising a V.sub.L comprising SEQ ID NO:22. In
certain embodiments, the antibody comprises: (i) a first heavy
chain (HC) comprising a V.sub.H comprising SEQ ID NO:24 and the
first Fc polypeptide comprising SEQ ID NO:70; (ii) a second heavy
chain (HC) comprising a V.sub.H comprising SEQ ID NO:24 and the
second Fc polypeptide comprising SEQ ID NO:63; and (iii) two light
chains each comprising a V.sub.L comprising SEQ ID NO:22. In
particular embodiments, the antibody comprises: (i) a first heavy
chain (HC) that comprises or consists of the amino acid sequence
set forth in SEQ ID NO:51; (ii) a second HC that comprises or
consists of the amino acid sequence set forth in SEQ ID NO:53; and
(iii) a first and a second light chain (LC) that each comprises or
consists of the amino acid sequence set forth in SEQ ID NO:54. In
certain embodiments, the antibody comprises: (i) a first heavy
chain (HC) that comprises or consists of the amino acid sequence
set forth in SEQ ID NO:71; (ii) a second HC that comprises or
consists of the amino acid sequence set forth in SEQ ID NO:73; and
(iii) a first and a second light chain (LC) that each comprises or
consists of the amino acid sequence set forth in SEQ ID NO:54.
[0056] In another aspect, the disclosure provides an isolated
antibody that specifically binds to a human triggering receptor
expressed on myeloid cells 2 (TREM2), wherein the antibody
comprises: (i) a first heavy chain (HC) comprising a V.sub.H
comprising SEQ ID NO:24 and the first Fc polypeptide comprising SEQ
ID NO:41; (ii) a second heavy chain (HC) comprising a V.sub.H
comprising SEQ ID NO:24 and the second Fc polypeptide comprising
SEQ ID NO:39; and (iii) two light chains each comprising a V.sub.L
comprising SEQ ID NO:22. In some embodiments, the antibody
comprises: (i) a first heavy chain (HC) that comprises or consists
of the amino acid sequence set forth in SEQ ID NO:42; (ii) a second
HC that comprises or consists of the amino acid sequence set forth
in SEQ ID NO:53; and (iii) a first and a second light chain (LC)
that each comprises or consists of the amino acid sequence set
forth in SEQ ID NO:54.
[0057] In another aspect, the disclosure provides an isolated
antibody that specifically binds to a human triggering receptor
expressed on myeloid cells 2 (TREM2), wherein the antibody
comprises: (i) a first heavy chain (HC) comprising a V.sub.H
comprising SEQ ID NO:24 and the first Fc polypeptide comprising SEQ
ID NO:64; (ii) a second heavy chain (HC) comprising a V.sub.H
comprising SEQ ID NO:24 and the second Fc polypeptide comprising
SEQ ID NO:63; and (iii) two light chains each comprising a V.sub.L
comprising SEQ ID NO:22. In some embodiments, the antibody
comprises: (i) a first heavy chain (HC) that comprises or consists
of the amino acid sequence set forth in SEQ ID NO:65; (ii) a second
HC that comprises or consists of the amino acid sequence set forth
in SEQ ID NO:73; and (iii) a first and a second light chain (LC)
that each comprises or consists of the amino acid sequence set
forth in SEQ ID NO:54.
[0058] In another aspect, the disclosure provides an isolated
antibody that specifically binds to a human triggering receptor
expressed on myeloid cells 2 (TREM2), wherein the antibody
comprises: (i) a first heavy chain (HC) comprising a V.sub.H
comprising SEQ ID NO:24 and the first Fc polypeptide comprising SEQ
ID NO:44; (ii) a second heavy chain (HC) comprising a V.sub.H
comprising SEQ ID NO:24 and the second Fc polypeptide comprising
SEQ ID NO:39; and (iii) two light chains each comprising a V.sub.L
comprising SEQ ID NO:22. In some embodiments, the antibody
comprises: (i) a first heavy chain (HC) that comprises or consists
of the amino acid sequence set forth in SEQ ID NO:45; (ii) a second
HC that comprises or consists of the amino acid sequence set forth
in SEQ ID NO:53; and (iii) a first and a second light chain (LC)
that each comprises or consists of the amino acid sequence set
forth in SEQ ID NO:54.
[0059] In another aspect, the disclosure provides an isolated
antibody that specifically binds to a human triggering receptor
expressed on myeloid cells 2 (TREM2), wherein the antibody
comprises: (i) a first heavy chain (HC) comprising a V.sub.H
comprising SEQ ID NO:24 and the first Fc polypeptide comprising SEQ
ID NO:66; (ii) a second heavy chain (HC) comprising a V.sub.H
comprising SEQ ID NO:24 and the second Fc polypeptide comprising
SEQ ID NO:63; and (iii) two light chains each comprising a V.sub.L
comprising SEQ ID NO:22. In some embodiments, the antibody
comprises: (i) a first heavy chain (HC) that comprises or consists
of the amino acid sequence set forth in SEQ ID NO:67; (ii) a second
HC that comprises or consists of the amino acid sequence set forth
in SEQ ID NO:73; and (iii) a first and a second light chain (LC)
that each comprises or consists of the amino acid sequence set
forth in SEQ ID NO:54.
[0060] In another aspect, the disclosure provides an isolated
antibody that specifically binds to a human triggering receptor
expressed on myeloid cells 2 (TREM2), wherein the antibody
comprises: (i) a first heavy chain (HC) comprising a V.sub.H
comprising SEQ ID NO:24 and the first Fc polypeptide comprising SEQ
ID NO:47; (ii) a second heavy chain (HC) comprising a V.sub.H
comprising SEQ ID NO:24 and the second Fc polypeptide comprising
SEQ ID NO:39; and (iii) two light chains each comprising a V.sub.L
comprising SEQ ID NO:22. In some embodiments, the antibody
comprises: (i) a first heavy chain (HC) that comprises or consists
of the amino acid sequence set forth in SEQ ID NO:48; (ii) a second
HC that comprises or consists of the amino acid sequence set forth
in SEQ ID NO:53; and (iii) a first and a second light chain (LC)
that each comprises or consists of the amino acid sequence set
forth in SEQ ID NO:54.
[0061] In another aspect, the disclosure provides an isolated
antibody that specifically binds to a human triggering receptor
expressed on myeloid cells 2 (TREM2), wherein the antibody
comprises: (i) a first heavy chain (HC) comprising a V.sub.H
comprising SEQ ID NO:24 and the first Fc polypeptide comprising SEQ
ID NO:68; (ii) a second heavy chain (HC) comprising a V.sub.H
comprising SEQ ID NO:24 and the second Fc polypeptide comprising
SEQ ID NO:63; and (iii) two light chains each comprising a V.sub.L
comprising SEQ ID NO:22. In some embodiments, the antibody
comprises: (i) a first heavy chain (HC) that comprises or consists
of the amino acid sequence set forth in SEQ ID NO:69; (ii) a second
HC that comprises or consists of the amino acid sequence set forth
in SEQ ID NO:73; and (iii) a first and a second light chain (LC)
that each comprises or consists of the amino acid sequence set
forth in SEQ ID NO:54.
[0062] In another aspect, the disclosure provides an isolated
antibody that specifically binds to a human triggering receptor
expressed on myeloid cells 2 (TREM2), wherein the antibody
comprises: (i) a first heavy chain (HC) comprising a V.sub.H
comprising SEQ ID NO:24 and the first Fc polypeptide comprising SEQ
ID NO:47 (ii) a second heavy chain (HC) comprising a V.sub.H
comprising SEQ ID NO:24 and the second Fc polypeptide comprising
SEQ ID NO:61; and (iii) two light chains each comprising a V.sub.L
comprising SEQ ID NO:22. In some embodiments, (i) a first heavy
chain (HC) that comprises or consists of the amino acid sequence
set forth in SEQ ID NO:48; (ii) a second HC that comprises or
consists of the amino acid sequence set forth in SEQ ID NO:52; and
(iii) a first and a second light chain (LC) that each comprises or
consists of the amino acid sequence set forth in SEQ ID NO:54.
[0063] In another aspect, the disclosure provides an isolated
antibody that specifically binds to a human triggering receptor
expressed on myeloid cells 2 (TREM2), wherein the antibody
comprises: (i) a first heavy chain (HC) comprising a V.sub.H
comprising SEQ ID NO:24 and the first Fc polypeptide comprising SEQ
ID NO:68; (ii) a second heavy chain (HC) comprising a V.sub.H
comprising SEQ ID NO:24 and the second Fc polypeptide comprising
SEQ ID NO:61; and (iii) two light chains each comprising a V.sub.L
comprising SEQ ID NO:22. In some embodiments, the antibody
comprises: (i) a first heavy chain (HC) that comprises or consists
of the amino acid sequence set forth in SEQ ID NO:69; (ii) a second
HC that comprises or consists of the amino acid sequence set forth
in SEQ ID NO:72; and (iii) a first and a second light chain (LC)
that each comprises or consists of the amino acid sequence set
forth in SEQ ID NO:54.
[0064] In another aspect, the disclosure provides an isolated
antibody that specifically binds to a human triggering receptor
expressed on myeloid cells 2 (TREM2), wherein the antibody
comprises: (i) a first heavy chain (HC) comprising a V.sub.H
comprising SEQ ID NO:24 and the first Fc polypeptide comprising SEQ
ID NO:50; (ii) a second heavy chain (HC) comprising a V.sub.H
comprising SEQ ID NO:24 and the second Fc polypeptide comprising
SEQ ID NO:39; and (iii) two light chains each comprising a V.sub.L
comprising SEQ ID NO:22. In some embodiments, the antibody
comprises: (i) a first heavy chain (HC) that comprises or consists
of the amino acid sequence set forth in SEQ ID NO:51; (ii) a second
HC that comprises or consists of the amino acid sequence set forth
in SEQ ID NO:53; and (iii) a first and a second light chain (LC)
that each comprises or consists of the amino acid sequence set
forth in SEQ ID NO:54.
[0065] In another aspect, the disclosure provides an isolated
antibody that specifically binds to a human triggering receptor
expressed on myeloid cells 2 (TREM2), wherein the antibody
comprises: (i) a first heavy chain (HC) comprising a V.sub.H
comprising SEQ ID NO:24 and the first Fc polypeptide comprising SEQ
ID NO:70; (ii) a second heavy chain (HC) comprising a V.sub.H
comprising SEQ ID NO:24 and the second Fc polypeptide comprising
SEQ ID NO:63; and (iii) two light chains each comprising a V.sub.L
comprising SEQ ID NO:22. In some embodiments, the antibody
comprises: (i) a first heavy chain (HC) that comprises or consists
of the amino acid sequence set forth in SEQ ID NO:71; (ii) a second
HC that comprises or consists of the amino acid sequence set forth
in SEQ ID NO:73; and (iii) a first and a second light chain (LC)
that each comprises or consists of the amino acid sequence set
forth in SEQ ID NO:54.
[0066] In some embodiments of any of the aspects described herein,
the antibody decreases levels of soluble TREM2 protein (sTREM2). In
some embodiments, the antibody enhances TREM2 activity. In some
embodiments, the antibody enhances phagocytosis or enhances the
migration, differentiation, function, or survival of myeloid cells,
microglia, or macrophages. In some embodiments, the antibody
enhances microglia function without increasing neuroinflammation.
In some embodiments, the antibody enhances Syk phosphorylation. In
some embodiments, the antibody enhances Syk phosphorylation in the
presence of a TREM2 ligand. In some embodiments, the antibody
exhibits cross-reactivity with a cynomolgus TREM2 protein.
[0067] In another aspect, the disclosure provides a pharmaceutical
composition comprising the isolated antibody described herein and a
pharmaceutically acceptable carrier.
[0068] In another aspect, the disclosure provides a kit comprising:
the isolated antibody described herein or the pharmaceutical
composition described herein; and instructions for use thereof.
[0069] In another aspect, the disclosure provides a method of
treating a neurodegenerative disease in a subject, comprising
administering to the subject the isolated antibody described herein
or the pharmaceutical composition described herein. In some
embodiments, the neurodegenerative disease is selected from the
group consisting of. Alzheimer's disease, primary age-related
tauopathy, progressive supranuclear palsy (PSP), frontotemporal
dementia, frontotemporal dementia with parkinsonism linked to
chromosome 17, argyrophilic grain dementia, amyotrophic lateral
sclerosis, amyotrophic lateral sclerosis/parkinsonism-dementia
complex of Guam (ALS-PDC), corticobasal degeneration, chronic
traumatic encephalopathy, Creutzfeldt-Jakob disease, dementia
pugilistica, diffuse neurofibrillary tangles with calcification,
Down's syndrome, familial British dementia, familial Danish
dementia, Gerstmann-Straussler-Scheinker disease, globular glial
tauopathy, Guadeloupean parkinsonism with dementia, Guadelopean
PSP, Hallevorden-Spatz disease, hereditary diffuse
leukoencephalopathy with spheroids (HDLS), Huntington's disease,
inclusion-body myositis, multiple system atrophy, myotonic
dystrophy, Nasu-Hakola disease, neurofibrillary tangle-predominant
dementia, Niemann-Pick disease type C, pallido-ponto-nigral
degeneration, Parkinson's disease, Pick's disease, postencephalitic
parkinsonism, prion protein cerebral amyloid angiopathy,
progressive subcortical gliosis, subacute sclerosing
panencephalitis, and tangle only dementia.
[0070] In another aspect, the disclosure provides a method of
decreasing levels of sTREM2 in a subject having a neurodegenerative
disease, comprising administering to the subject the isolated
antibody described herein or the pharmaceutical composition
described herein.
[0071] In another aspect, the disclosure provides a method of
enhancing TREM2 activity in a subject having a neurodegenerative
disease, comprising administering to the subject the isolated
antibody described herein or the pharmaceutical composition
described herein.
[0072] In another aspect, the disclosure provides an isolated
polynucleotide comprising a nucleotide sequence encoding the
antibody described herein.
[0073] In another aspect, the disclosure provides an isolated
polynucleotide comprising a nucleotide sequence encoding any one of
SEQ ID NOS:42, 45, 48, 51, 53, 54, and 61.
[0074] In another aspect, the disclosure provides an isolated
polynucleotide comprising a nucleotide sequence encoding SEQ ID
NOS:42, 53, and 54.
[0075] In another aspect, the disclosure provides an isolated
polynucleotide comprising a nucleotide sequence encoding SEQ ID
NOS:45, 53, and 54.
[0076] In another aspect, the disclosure provides an isolated
polynucleotide comprising a nucleotide sequence encoding SEQ ID
NOS:48, 53, and 54.
[0077] In another aspect, the disclosure provides an isolated
polynucleotide comprising a nucleotide sequence encoding SEQ ID
NOS:48, 52, and 54.
[0078] In another aspect, the disclosure provides an isolated
polynucleotide comprising a nucleotide sequence encoding SEQ ID
NOS:51, 53, and 54.
[0079] In another aspect, the disclosure provides a vector
comprising the polynucleotide described herein.
[0080] In another aspect, the disclosure provides a host cell
comprising the polynucleotide described herein or the vector
described herein.
[0081] In another aspect, the disclosure provides a method of
expressing an antibody that specifically binds to a human
triggering receptor expressed on myeloid cells 2 (TREM2),
comprising: culturing the host cell described herein in under
conditions suitable for expression of the antibody.
BRIEF DESCRIPTION OF THE DRAWINGS
[0082] FIGS. 1A-1H include dose-titrated cell binding curves in
human TREM2-Dap12 overexpressing HEK cells for humanized and
sequence optimized variants of a representative anti-TREM2 antibody
(CL0020188).
[0083] FIG. 2 includes dose-response binding curves to human TREM2
in HEK 293 cells for a representative ATV:TREM2 and corresponding
anti-TREM2 antibody with non-transferrin-binding Fc
("anti-TREM2").
[0084] FIG. 3 includes dose-response curves of pSyk signal
activation by a representative ATV:TREM2 and corresponding
anti-TREM2 antibody (TREM2 IgG) in TREM2-expressing HEK 293
cells.
[0085] FIGS. 4A and 4B include dose-response curves of lipid
clearance in iPSC microglia in response to treatment with a
representative ATV:TREM2.
[0086] FIGS. 5A and 5B show representative images of lipid
accumulation in iPSC-derived microglial cells treated with
ATV:TREM2 after oleic acid challenge (FIG. 5A) with quantification
of lipid accumulation in treated cells (FIG. 5B).
[0087] FIG. 5C is a heat map illustrating the modulation of levels
of triglyceride, acyl carnitine, and TCA cycle intermediate species
in iPSC-derived microglial cells treated with ATV:TREM2 after
myelin challenge.
[0088] FIGS. 5D-5F include bar graphs illustrating the change in
levels of representative triglyceride, acyl carnitine, and TCA
cycle intermediate species levels in iPSC-derived microglial cells
treated with ATV:TREM2 after myelin challenge.
[0089] FIGS. 6A-6C include bar graphs illustrating the change in
levels of specific triglyceride, ceramide, and acyl carnitine
species in iPSC-derived microglial cells treated with ATV:TREM2
after myelin challenge.
[0090] FIG. 7A includes representative images from a Western blot
of mTOR signal pathway targets in iPSC-derived microglia treated
with ATV:TREM2.
[0091] FIGS. 7B-7E include plots illustrating the change in levels
of mTOR signal pathway targets in iPSC-derived microglia treated
with ATV:TREM2.
[0092] FIG. 8 is a bar graph illustrating the change in levels of
progranulin (PGRN) in iPSC-derived microglia treated with
ATV:TREM2.
[0093] FIG. 9 is a bar graph illustrating the change in levels of a
representative bis(monoacylglycero)phosphate (BMP) species in
iPSC-derived microglia treated with ATV:TREM2.
[0094] FIG. 10A is a representative kinetic graph of oxygen
consumption in iPSC-derived microglial cells treated with
ATV:TREM2.
[0095] FIG. 10B is a bar graph illustrating the maximal respiratory
capacity of iPSC-derived microglial cells treated with ATV:TREM2 in
the presence and absence of a CPT1 inhibitor.
[0096] FIG. 11A is a dose-response curve for cell viability in
human macrophage cells treated with anti-TREM2 antibodies.
[0097] FIGS. 11B and 11C include bar graphs illustrating EC50 and
Emax for the dose-response curves in FIG. 11A.
[0098] FIG. 12 is a heat map of relative cytokine release in human
macrophage cells treated with anti-TREM2 antibodies.
[0099] FIGS. 13A-13C include volcano plots that illustrate relative
changes in triglyceride species in iPSC-derived microglial cells
treated with anti-TREM2 antibodies.
[0100] FIGS. 13D and 13E include bar graphs showing the change in
levels of representative triglyceride species in iPSC-derived
microglial cells treated with anti-TREM2 antibodies.
[0101] FIG. 14A is a plot of TREM2 level as a function of antibody
concentration in the cell lysate of iPSC-derived microglial cells
treated with anti-TREM2 antibodies.
[0102] FIG. 14B is a plot of TREM2 level as a function of antibody
concentration in the cell culture media of iPSC-derived microglial
cells treated with anti-TREM2 antibodies.
[0103] FIG. 15 is a plot illustrating the pharmacokinetic profile
of anti-TREM2 antibodies dosed in cynomolgus monkeys.
[0104] FIGS. 16A and 16B are plots showing EdU.sup.+Iba.sup.+ cells
per mm.sup.2 (FIG. 16A) and relative Iba.sup.+ area (FIG. 16B) in
brains of TB36/hTfR KI mice treated with either ATV:TREM2 or
ATV:RSV.
[0105] FIGS. 17A and 17B are plots showing EdU.sup.+Iba.sup.+ cells
per mm.sup.2 (FIG. 17A) and relative Iba.sup.+ area (FIG. 17B) in
brains of TB36/hTfR KI mice treated with either ATV:TREM2, a
corresponding TREM2 antibody, reference antibody #2, or ATV:RSV.
Graphs display mean.+-.SEM and p values: oneway ANOVA with Tukey's
multiple comparison test; * p.ltoreq.0.05, ** p.ltoreq.0.01, ***
p.ltoreq.0.001, **** p.ltoreq.0.0001.
[0106] FIGS. 18A and 18B are plots showing cytokines IP-10 (FIG.
18A) and MCP-5 (FIG. 18B) levels in brains of TB36/hTfR KI mice
treated with either ATV:TREM2, a corresponding TREM2 antibody,
reference antibody #2, or ATV:RSV. Graphs display mean values
across experimental replicates .+-.SEM. Graphs display mean.+-.SEM
and p values: one-way ANOVA with Tukey's multiple comparison test;
** p.ltoreq.0.01, *** p.ltoreq.0.001, **** p.ltoreq.0.0001.
[0107] FIG. 19 is a plot showing glial marker CSF1R level in brains
of TB36/hTfR KI mice treated with either ATV:TREM2, a corresponding
TREM2 antibody, reference antibody #2, or ATV:RSV. Graphs display
mean.+-.SEM and p values: one-way ANOVA with Tukey's multiple
comparison test; * p.ltoreq.0.05, ** p.ltoreq.0.01,
*p.ltoreq.0.001, **** p.ltoreq.0.0001.
[0108] FIG. 20 is a plot showing the plasma PK profile of either
ATV:TREM2, a corresponding TREM2 antibody, reference antibody #2,
or ATV:RSV in TB36/hTfR KI mice.
[0109] FIG. 21 is a plot showing the brain PK profile of either
ATV:TREM2, a corresponding TREM2 antibody, reference antibody #2,
or ATV:RSV in TB36/hTfR KI mice.
DETAILED DESCRIPTION
I. Introduction
[0110] TREM2 is a transmembrane receptor that is expressed on the
cell surface of microglia, dendritic cells, macrophages, and
osteoclasts. Without being bound to a particular theory, it is
believed that upon ligand binding, TREM2 forms a signaling complex
with a transmembrane adapter protein, DNAX-activating protein 12
(DAP12), which in turn is tyrosine phosphorylated by the protein
kinase SRC. It is believed that the activated TREM2/DAP12 signaling
complex mediates intracellular signaling by recruiting and
phosphorylating kinases such as Syk kinase. TREM2/DAP12 signaling
modulates activities such as phagocytosis, cell growth and
survival, pro-inflammatory cytokine secretion, and the migration of
cells such as microglia and macrophages. TREM2 undergoes regulated
intramembrane proteolysis, in which the membrane-associated
full-length TREM2 is cleaved by the metalloprotease ADAM10 into a
sTREM2 portion that is shed from the cell and a membrane-retained
C-terminal fragment that is further degraded by a gamma-secretase.
Altered levels of sTREM2 have been reported in patients having
Alzheimer's disease or frontotemporal dementia and having a
mutation in TREM2. Additionally, mutations in TREM2 are associated
with altered functions such as impaired phagocytosis and reduced
microglial function.
[0111] As detailed in the Examples section below, antibodies have
been generated that specifically bind to human TREM2 and that
modulate one or more downstream functions of the TREM2/DAP12
signaling complex. In certain embodiments, the antibodies further
comprise an Fc polypeptide containing mutations that permit binding
of the Fc polypeptide to a transferrin receptor (TfR from, e.g., a
human). In some aspects, the antibodies disclosed herein are able
to bind, through the modified Fc polypeptide, to a transferrin
receptor protein (e.g., expressed on the surface of a brain
endothelial cell (BECs)) and can thereby cross the blood-brain
barrier (BBB) more effectively than antibodies lacking the
TfR-binding Fc mutations. In certain embodiments, the antibodies
disclosed herein comprise mutations in an Fc polypeptide that
reduce or eliminate effector function and mutations that increase
in vivo half-life, e.g., by increasing binding of antibody Fc to Fc
neonatal receptor (FcRn).
[0112] In some embodiments, the anti-TREM2 antibodies enhance TREM2
activity, e.g., enhance phagocytosis or enhance the
differentiation, function, migration, or survival of myeloid cells,
microglia, or macrophages. Thus, in another aspect, methods of
enhancing TREM2 activity, e.g., in a subject having a
neurodegenerative disease, are provided.
[0113] In some embodiments, the anti-TREM2 antibodies reduce
shedding of sTREM2. Thus, in another aspect, methods of decreasing
levels of sTREM2, e.g., in a subject having a neurodegenerative
disease, are provided.
II. Definitions
[0114] As used herein, the singular forms "a," "an" and "the"
include plural referents unless the content clearly dictates
otherwise. Thus, for example, reference to "an antibody" optionally
includes a combination of two or more such molecules, and the
like.
[0115] As used herein, the terms "about" and "approximately," when
used to modify an amount specified in a numeric value or range,
indicate that the numeric value as well as reasonable deviations
from the value known to the skilled person in the art, for example
20%, +10%, or +5%, are within the intended meaning of the recited
value.
[0116] As used herein, the term "TREM2 protein" refers to a
triggering receptor expressed on myeloid cells 2 protein that is
encoded by the gene TREM2. As used herein, a "TREM2 protein" refers
to a native (i.e., wild-type) TREM2 protein of any vertebrate, such
as but not limited to human, non-human primates (e.g., cynomolgus
monkey), rodents (e.g., mice, rat), and other mammals. In some
embodiments, a TREM2 protein is a human TREM2 protein having the
sequence identified in UniprotKB accession number Q9NZC2 (SEQ ID
NO:1).
[0117] As used herein, the term "anti-TREM2 antibody" refers to an
antibody that specifically binds to a TREM2 protein (e.g., human
TREM2).
[0118] As used herein, the term "antibody" refers to a protein with
an immunoglobulin fold that specifically binds to an antigen via
its variable regions. The term encompasses intact polyclonal
antibodies, intact monoclonal antibodies, single chain antibodies,
multispecific antibodies such as bispecific antibodies,
monospecific antibodies, monovalent antibodies, chimeric
antibodies, humanized antibodies, and human antibodies. The term
"antibody," as used herein, also includes antibody fragments that
retain binding specificity via its variable regions, including but
not limited to Fab, F(ab').sub.2, Fv, scFv, and bivalent scFv.
Antibodies can contain light chains that are classified as either
kappa or lambda. Antibodies can contain heavy chains that are
classified as gamma, mu, alpha, delta, or epsilon, which in turn
define the immunoglobulin classes, IgG, IgM, IgA, IgD and IgE,
respectively.
[0119] As used herein, the term "anti-TREM2 antigen binding
portion" refers to an antigen binding segment or entity that
specifically binds to a TREM2 protein (e.g., human TREM2). The
terms "antigen-binding portion" and "antigen-binding fragment" are
used interchangeably herein and refer to one or more fragments of
an antibody that retains the ability to specifically bind to an
antigen (e.g., a TREM2 protein) via its variable region. Examples
of antigen-binding fragments include, but are not limited to, a Fab
fragment (a monovalent fragment consisting of the V.sub.L, V.sub.H,
CL and CH1 domains), F(ab').sub.2 fragment (a bivalent fragment
comprising two Fab fragments linked by a disulfide bridge at the
hinge region), single chain Fv (scFv), disulfide-linked Fv (dsFv),
complementarity determining regions (CDRs), a V.sub.L (light chain
variable region), and a V.sub.H (heavy chain variable region).
[0120] The term "variable region" or "variable domain" refers to a
domain in an antibody heavy chain or light chain that is derived
from a germline Variable (V) gene, Diversity (D) gene, or Joining
(J) gene (and not derived from a Constant (C and CS) gene segment),
and that gives an antibody its specificity for binding to an
antigen. Typically, an antibody variable region comprises four
conserved "framework" regions interspersed with three hypervariable
"complementarity determining regions."
[0121] The term "complementarity determining region" or "CDR"
refers to the three hypervariable regions in each chain that
interrupt the four framework regions established by the light and
heavy chain variable regions. The CDRs are primarily responsible
for antibody binding to an epitope of an antigen. The CDRs of each
chain are typically referred to as CDR1, CDR2, and CDR3, numbered
sequentially starting from the N-terminus, and are also typically
identified by the chain in which the particular CDR is located.
Thus, a V.sub.H CDR3 or CDR-H3 is located in the variable region of
the heavy chain of the antibody in which it is found, whereas a
V.sub.L CDR1 or CDR-L1 is the CDR1 from the variable region of the
light chain of the antibody in which it is found.
[0122] The "framework regions" or "FRs" of different light or heavy
chains are relatively conserved within a species. The framework
region of an antibody, that is the combined framework regions of
the constituent light and heavy chains, serves to position and
align the CDRs in three-dimensional space. Framework sequences can
be obtained from public DNA databases or published references that
include germline antibody gene sequences. For example, germline DNA
sequences for human heavy and light chain variable region genes can
be found in the "VBASE2" germline variable gene sequence database
for human and mouse sequences.
[0123] The amino acid sequences of the CDRs and framework regions
can be determined using various well-known definitions in the art,
e.g., Kabat, Chothia, international ImMunoGeneTics database (IMGT),
AbM, and observed antigen contacts ("Contact"). In some
embodiments, CDRs are determined according to the Contact
definition. See, MacCallum et al., J. Mol. Biol., 262:732-745
(1996). In some embodiments, CDRs are determined by a combination
of Kabat, Chothia, and/or Contact CDR definitions.
[0124] The term "epitope" refers to the area or region of an
antigen to which the CDRs of an antibody specifically binds and can
include a few amino acids or portions of a few amino acids, e.g., 5
or 6, or more, e.g., 20 or more amino acids, or portions of those
amino acids. For example, where the target is a protein, the
epitope can be comprised of consecutive amino acids (e.g., a linear
epitope), or amino acids from different parts of the protein that
are brought into proximity by protein folding (e.g., a
discontinuous or conformational epitope). In some embodiments, the
epitope is phosphorylated at one amino acid (e.g., at a serine or
threonine residue).
[0125] As used herein, the phrase "recognizes an epitope," as used
with reference to an anti-TREM2 antibody, means that the antibody
CDRs interact with or specifically bind to the antigen (i.e., the
TREM2 protein) at that epitope or a portion of the antigen
containing that epitope.
[0126] A "monoclonal antibody" refers to antibodies produced by a
single clone of cells or a single cell line and consisting of or
consisting essentially of antibody molecules that are identical in
their primary amino acid sequence.
[0127] A "polyclonal antibody" refers to an antibody obtained from
a heterogeneous population of antibodies in which different
antibodies in the population bind to different epitopes of an
antigen.
[0128] A "chimeric antibody" refers to an antibody molecule in
which the constant region, or a portion thereof, is altered,
replaced or exchanged so that the antigen-binding site (i.e.,
variable region, CDR, or portion thereof) is linked to a constant
region of a different or altered class, effector function and/or
species, or in which the variable region, or a portion thereof, is
altered, replaced or exchanged with a variable region having a
different or altered antigen specificity (e.g., CDR and framework
regions from different species). In some embodiments, a chimeric
antibody is a monoclonal antibody comprising a variable region from
one source or species (e.g., mouse) and a constant region derived
from a second source or species (e.g., human). Methods for
producing chimeric antibodies are described in the art.
[0129] A "humanized antibody" is a chimeric immunoglobulin derived
from a non-human source (e.g., murine) that contains minimal
sequences derived from the non-human immunoglobulin outside the
CDRs. In general, a humanized antibody will comprise at least one
(e.g., two) antigen-binding variable domain(s), in which the CDR
regions substantially correspond to those of the non-human
immunoglobulin and the framework regions substantially correspond
to those of a human immunoglobulin sequence. The humanized antibody
can also comprise at least a portion of an immunoglobulin constant
region (Fc), typically that of a human immunoglobulin sequence.
Methods of antibody humanization are known in the art.
[0130] A "human antibody" or a "fully human antibody" is an
antibody having human heavy chain and light chain sequences,
typically derived from human germline genes. In some embodiments,
the antibody is produced by a human cell, by a non-human animal
that utilizes human antibody repertoires (e.g., transgenic mice
that are genetically engineered to express human antibody
sequences), or by phage display platforms.
[0131] The term "specifically binds" refers to a molecule (e.g., an
antibody or an antigen-binding portion thereof) that binds to an
epitope or target with greater affinity, greater avidity, and/or
greater duration to that epitope or target in a sample than it
binds to another epitope or non-target compound (e.g., a
structurally different antigen). In some embodiments, an antibody
(or an antigen-binding portion thereof) that specifically binds to
an epitope or target is an antibody (or an antigen-binding portion
thereof) that binds to the epitope or target with at least 5-fold
greater affinity than other epitopes or non-target compounds, e.g.,
at least 5-fold, 10-fold, 100-fold, 1,000-fold, 10,000-fold, or
greater affinity. The term "specific binding," "specifically binds
to," or "is specific for" a particular epitope or target, as used
herein, can be exhibited, for example, by a molecule having an
equilibrium dissociation constant K.sub.D for the epitope or target
to which it binds of, e.g., 10.sup.-4 M or smaller, e.g., 10.sup.-5
M, 10.sup.-6 M, 10.sup.-7 M, 10.sup.-8 M, 10.sup.-9 M, 10.sup.-10
M, 10.sup.-11 M, or 10.sup.-12 M. It will be recognized by one of
skill that an antibody that specifically binds to a target (e.g., a
TREM2 protein) from one species may also specifically bind to
orthologs of that target (e.g., the TREM2 protein).
[0132] The term "binding affinity" is used herein to refer to the
strength of a non-covalent interaction between two molecules, e.g.,
between an antibody (or an antigen-binding portion thereof) and an
antigen. Thus, for example, the term may refer to 1:1 interactions
between an antibody (or an antigen-binding portion thereof) and an
antigen, unless otherwise indicated or clear from context. Binding
affinity may be quantified by measuring an equilibrium dissociation
constant (K.sub.D), which refers to the dissociation rate constant
(k.sub.d, time.sup.-1) divided by the association rate constant
(k.sub.a, time.sup.-1 M.sup.-1). K.sub.D can be determined by
measurement of the kinetics of complex formation and dissociation,
e.g., using Surface Plasmon Resonance (SPR) methods, e.g., a
Biacore.TM. system; kinetic exclusion assays such as KinExA.RTM.;
and BioLayer interferometry (e.g., using the ForteBio.RTM. Octet
platform). As used herein, "binding affinity" includes not only
formal binding affinities, such as those reflecting 1:1
interactions between an antibody (or an antigen-binding portion
thereof) and an antigen, but also apparent affinities for which
K.sub.D values are calculated that may reflect avid binding.
[0133] The term "cross-reacts," as used herein, refers to the
ability of an antibody to bind to an antigen other than the antigen
against which the antibody was raised. In some embodiments,
cross-reactivity refers to the ability of an antibody to bind to an
antigen from another species than the antigen against which the
antibody was raised. As a non-limiting example, an anti-TREM2
antibody as described herein that is raised against a human TREM2
peptide can exhibit cross-reactivity with a TREM2 peptide or
protein from a different species (e.g., monkey or mouse).
[0134] A "transferrin receptor" or "TfR" as used herein refers to
transferrin receptor protein 1. The human transferrin receptor 1
polypeptide sequence is set forth in SEQ ID NO:62. Transferrin
receptor protein 1 sequences from other species are also known
(e.g., chimpanzee, accession number XP_003310238.1; rhesus monkey,
NP_001244232.1; dog, NP_001003111.1; cattle, NP_001193506.1; mouse,
NP_035768.1; rat, NP_073203.1; and chicken, NP_990587.1). The term
"transferrin receptor" also encompasses allelic variants of
exemplary reference sequences, e.g., human sequences, that are
encoded by a gene at a transferrin receptor protein 1 chromosomal
locus. Full length transferrin receptor protein includes a short
N-terminal intracellular region, a transmembrane region, and a
large extracellular domain. The extracellular domain is
characterized by three domains: a protease-like domain, a helical
domain, and an apical domain. The apical domain sequence of human
transferrin receptor 1 is set forth in SEQ ID NO:55.
[0135] The terms "CH3 domain" and "CH2 domain" as used herein refer
to immunoglobulin constant region domain polypeptides. In the
context of IgG antibodies, a CH3 domain polypeptide refers to the
segment of amino acids from about position 341 to about position
447 as numbered according to the EU numbering scheme, and a CH2
domain polypeptide refers to the segment of amino acids from about
position 231 to about position 340 as numbered according to the EU
numbering scheme. CH2 and CH3 domain polypeptides may also be
numbered by the IMGT (ImMunoGeneTics) numbering scheme in which the
CH2 domain numbering is 1-110 and the CH3 domain numbering is
1-107, according to the IMGT Scientific chart numbering (IMGT
website). CH2 and CH3 domains are part of the Fc region of an
immunoglobulin. In the context of IgG antibodies, an Fc region
refers to the segment of amino acids from about position 231 to
about position 447 as numbered according to the EU numbering
scheme. As used herein, the term "Fc region" may also include at
least a part of a hinge region of an antibody. An exemplary partial
hinge region sequence is set forth in SEQ ID NO:57.
[0136] The terms "corresponding to," "determined with reference
to," or "numbered with reference to" when used in the context of
the identification of a given amino acid residue in a polypeptide
sequence, refers to the position of the residue of a specified
reference sequence when the given amino acid sequence is maximally
aligned and compared to the reference sequence. Thus, for example,
an amino acid residue in a polypeptide "corresponds to" an amino
acid in the region of SEQ ID NO:38 from amino acids 111-217 when
the residue aligns with the amino acid in SEQ ID NO:38 when
optimally aligned to SEQ ID NO:38. The polypeptide that is aligned
to the reference sequence need not be the same length as the
reference sequence.
[0137] As used herein, the term "Fc polypeptide" refers to the
C-terminal region of a naturally occurring immunoglobulin heavy
chain polypeptide that is characterized by an Ig fold as a
structural domain. An Fc polypeptide contains constant region
sequences including at least the CH2 domain and/or the CH3 domain
and may contain at least part of the hinge region, but does not
contain a variable region.
[0138] A "modified Fc polypeptide" refers to an Fc polypeptide that
has at least one mutation, e.g., a substitution, deletion or
insertion, as compared to a wild-type immunoglobulin heavy chain Fc
polypeptide sequence, but retains the overall Ig fold or structure
of the native Fc polypeptide.
[0139] The term "isolated," as used with reference to a nucleic
acid or protein (e.g., antibody), denotes that the nucleic acid or
protein is essentially free of other cellular components with which
it is associated in the natural state. Purity and homogeneity are
typically determined using analytical chemistry techniques such as
electrophoresis (e.g., polyacrylamide gel electrophoresis) or
chromatography (e.g., high performance liquid chromatography). In
some embodiments, an isolated nucleic acid or protein (e.g.,
antibody) is at least 85% pure, at least 90% pure, at least 95%
pure, or at least 99% pure.
[0140] The term "amino acid" refers to naturally occurring and
synthetic amino acids, as well as amino acid analogs and amino acid
mimetics that function in a manner similar to the naturally
occurring amino acids. Naturally occurring amino acids are those
encoded by the genetic code, as well as those amino acids that are
later modified, e.g., hydroxyproline, .gamma.-carboxyglutamate, and
O-phosphoserine. Naturally occurring .alpha.-amino acids include,
without limitation, alanine (Ala), cysteine (Cys), aspartic acid
(Asp), glutamic acid (Glu), phenylalanine (Phe), glycine (Gly),
histidine (His), isoleucine (Ile), arginine (Arg), lysine (Lys),
leucine (Leu), methionine (Met), asparagine (Asn), proline (Pro),
glutamine (Gln), serine (Ser), threonine (Thr), valine (Val),
tryptophan (Trp), tyrosine (Tyr), and combinations thereof.
Stereoisomers of a naturally occurring .alpha.-amino acids include,
without limitation, D-alanine (D-Ala), D-cysteine (D-Cys),
D-aspartic acid (D-Asp), D-glutamic acid (D-Glu), D-phenylalanine
(D-Phe), D-histidine (D-His), D-isoleucine (D-Ile), D-arginine
(D-Arg), D-lysine (D-Lys), D-leucine (D-Leu), D-methionine (D-Met),
D-asparagine (D-Asn), D-proline (D-Pro), D-glutamine (D-Gln),
D-serine (D-Ser), D-threonine (D-Thr), D-valine (D-Val),
D-tryptophan (D-Trp), D-tyrosine (D-Tyr), and combinations thereof
"Amino acid analogs" refer to compounds that have the same basic
chemical structure as a naturally occurring amino acid, i.e., an a
carbon that is bound to a hydrogen, a carboxyl group, an amino
group, and an R group, e.g., homoserine, norleucine, methionine
sulfoxide, methionine methyl sulfonium. Such analogs have modified
R groups (e.g., norleucine) or modified peptide backbones, but
retain the same basic chemical structure as a naturally occurring
amino acid. "Amino acid mimetics" refers to chemical compounds that
have a structure that is different from the general chemical
structure of an amino acid, but that functions in a manner similar
to a naturally occurring amino acid. Amino acids may be referred to
herein by either their commonly known three letter symbols or by
the one-letter symbols recommended by the IUPAC-IUB Biochemical
Nomenclature Commission.
[0141] The terms "polypeptide" and "peptide" are used
interchangeably herein to refer to a polymer of amino acid residues
in a single chain. The terms apply to amino acid polymers in which
one or more amino acid residue is an artificial chemical mimetic of
a corresponding naturally occurring amino acid, as well as to
naturally occurring amino acid polymers and non-naturally occurring
amino acid polymers. Amino acid polymers may comprise entirely
L-amino acids, entirely D-amino acids, or a mixture of L and D
amino acids.
[0142] The term "protein" as used herein refers to either a
polypeptide or a dimer (i.e., two) or multimer (i.e., three or
more) of single chain polypeptides. The single chain polypeptides
of a protein may be joined by a covalent bond, e.g., a disulfide
bond, or non-covalent interactions.
[0143] The terms "polynucleotide" and "nucleic acid"
interchangeably refer to chains of nucleotides of any length, and
include DNA and RNA. The nucleotides can be deoxyribonucleotides,
ribonucleotides, modified nucleotides or bases, and/or their
analogs, or any substrate that can be incorporated into a chain by
DNA or RNA polymerase. A polynucleotide may comprise modified
nucleotides, such as methylated nucleotides and their analogs.
Examples of polynucleotides contemplated herein include single- and
double-stranded DNA, single- and double-stranded RNA, and hybrid
molecules having mixtures of single- and double-stranded DNA and
RNA.
[0144] The terms "conservative substitution" and "conservative
mutation" refer to an alteration that results in the substitution
of an amino acid with another amino acid that can be categorized as
having a similar feature. Examples of categories of conservative
amino acid groups defined in this manner can include: a
"charged/polar group" including Glu (Glutamic acid or E), Asp
(Aspartic acid or D), Asn (Asparagine or N), Gln (Glutamine or Q),
Lys (Lysine or K), Arg (Arginine or R), and His (Histidine or H);
an "aromatic group" including Phe (Phenylalanine or F), Tyr
(Tyrosine or Y), Trp (Tryptophan or W), and (Histidine or H); and
an "aliphatic group" including Gly (Glycine or G), Ala (Alanine or
A), Val (Valine or V), Leu (Leucine or L), Ile (Isoleucine or I),
Met (Methionine or M), Ser (Serine or S), Thr (Threonine or T), and
Cys (Cysteine or C). Within each group, subgroups can also be
identified. For example, the group of charged or polar amino acids
can be sub-divided into sub-groups including: a "positively-charged
sub-group" comprising Lys, Arg and His; a "negatively-charged
sub-group" comprising Glu and Asp; and a "polar sub-group"
comprising Asn and Gln. In another example, the aromatic or cyclic
group can be sub-divided into sub-groups including: a "nitrogen
ring sub-group" comprising Pro, His and Trp; and a "phenyl
sub-group" comprising Phe and Tyr. In another further example, the
aliphatic group can be sub-divided into sub-groups, e.g., an
"aliphatic non-polar sub-group" comprising Val, Leu, Gly, and Ala;
and an "aliphatic slightly-polar sub-group" comprising Met, Ser,
Thr, and Cys. Examples of categories of conservative mutations
include amino acid substitutions of amino acids within the
sub-groups above, such as, but not limited to: Lys for Arg or vice
versa, such that a positive charge can be maintained; Glu for Asp
or vice versa, such that a negative charge can be maintained; Ser
for Thr or vice versa, such that a free --OH can be maintained; and
Gln for Asn or vice versa, such that a free --NH.sub.2 can be
maintained. In some embodiments, hydrophobic amino acids are
substituted for naturally occurring hydrophobic amino acid, e.g.,
in the active site, to preserve hydrophobicity.
[0145] The terms "identical" or percent "identity," in the context
of two or more polypeptide sequences, refer to two or more
sequences or subsequences that are the same or have a specified
percentage of amino acid residues, e.g., at least 60%, at least
65%, at least 70%, at least 75%, at least 80%, at least 85%, at
least 90%, or at least 95% or greater, that are identical over a
specified region when compared and aligned for maximum
correspondence over a comparison window, or designated region as
measured using a sequence comparison algorithm or by manual
alignment and visual inspection.
[0146] For sequence comparison of polypeptides, typically one amino
acid sequence acts as a reference sequence, to which a candidate
sequence is compared. Alignment can be performed using various
methods available to one of skill in the art, e.g., visual
alignment or using publicly available software using known
algorithms to achieve maximal alignment. Such programs include the
BLAST programs, ALIGN, ALIGN-2 (Genentech, South San Francisco,
Calif.) or Megalign (DNASTAR). The parameters employed for an
alignment to achieve maximal alignment can be determined by one of
skill in the art. For sequence comparison of polypeptide sequences
for purposes of this application, the BLASTP algorithm standard
protein BLAST for aligning two proteins sequence with the default
parameters is used.
[0147] The terms "subject," "individual," and "patient," as used
interchangeably herein, refer to a mammal, including but not
limited to humans, non-human primates, rodents (e.g., rats, mice,
and guinea pigs), rabbits, cows, pigs, horses, and other mammalian
species. In one embodiment, the subject, individual, or patient is
a human.
[0148] The terms "treating," "treatment," and the like are used
herein to generally mean obtaining a desired pharmacologic and/or
physiologic effect. "Treating" or "treatment" may refer to any
indicia of success in the treatment or amelioration of a
neurodegenerative disease (e.g., Alzheimer's disease or another
neurodegenerative disease described herein), including any
objective or subjective parameter such as abatement, remission,
improvement in patient survival, increase in survival time or rate,
diminishing of symptoms or making the disease more tolerable to the
patient, slowing in the rate of degeneration or decline, or
improving a patient's physical or mental well-being. The treatment
or amelioration of symptoms can be based on objective or subjective
parameters. The effect of treatment can be compared to an
individual or pool of individuals not receiving the treatment, or
to the same patient prior to treatment or at a different time
during treatment.
[0149] The term "pharmaceutically acceptable excipient" refers to a
non-active pharmaceutical ingredient that is biologically or
pharmacologically compatible for use in humans or animals, such as,
but not limited to a buffer, carrier, or preservative.
[0150] As used herein, a "therapeutic amount" or "therapeutically
effective amount" of an agent (e.g., an antibody as described
herein) is an amount of the agent that treats, alleviates, abates,
or reduces the severity of symptoms of a disease in a subject. A
"therapeutic amount" of an agent (e.g., an antibody as described
herein) may improve patient survival, increase survival time or
rate, diminish symptoms, make an injury, disease, or condition
(e.g., a neurodegenerative disease) more tolerable, slow the rate
of degeneration or decline, or improve a patient's physical or
mental well-being.
[0151] The term "administer" refers to a method of delivering
agents, compounds, or compositions to the desired site of
biological action. These methods include, but are not limited to,
topical delivery, parenteral delivery, intravenous delivery,
intradermal delivery, intramuscular delivery, intrathecal delivery,
colonic delivery, rectal delivery, or intraperitoneal delivery. In
one embodiment, an antibody as described herein is administered
intravenously.
[0152] The term "control" or "control value" refers to a reference
value or baseline value. Appropriate controls can be determined by
one skilled in the art. In some instances, control values can be
determined relative to a baseline within the same subject or
experiment, e.g., a measurement of sTREM2 taken prior to treatment
with an anti-TREM2 antibody can be a control value for a
post-treatment measurement of sTREM2 levels in the same subject. In
other instances, the control value can be determined relative to a
control subject (e.g., a healthy control or a disease control) or
an average value in a population of control subjects (e.g., healthy
controls or disease controls, e.g., a population of 10, 20, 50,
100, 200, 500, 1000 control subjects or more), e.g, a measurement
of a subject's level of sTREM2 either at baseline or after
treatment can be compared to a healthy control value.
III. Anti-TREM2 Antibodies
[0153] In one aspect, antibodies that specifically bind to a TREM2
protein are provided. In some embodiments, the antibody
specifically binds to a human TREM2 protein. In some embodiments,
an anti-TREM2 antibody is selective for TREM2 over other TREM-like
receptors (e.g., TREM1).
[0154] In some embodiments, an anti-TREM2 antibody is an antibody
that comprises one or more complementarity determining region
(CDR), heavy chain variable region, and/or light chain variable
region sequences as disclosed herein. In some embodiments, an
anti-TREM2 antibody comprises one or more CDR, heavy chain variable
region, and/or light chain variable region sequences as disclosed
herein and further comprises one or more functional characteristics
as disclosed herein, e.g., an antibody that enhances TREM2 activity
(e.g., enhances phagocytosis, or enhances the migration,
differentiation, function, or survival of a cell such as a myeloid
cell, microglia, or macrophage) or an antibody that decreases
levels of sTREM2. In some embodiments, the anti-TREM2 antibody
comprises Fc polypeptides that comprise one or more modifications
as described herein.
[0155] In some embodiments, the anti-TREM2 antibody is a chimeric
antibody. In some embodiments, the anti-TREM2 antibody is a
humanized and/or affinity matured antibody.
Anti-TREM2 Sequences
[0156] In some embodiments, a heavy chain sequence, or a portion
thereof, and/or a light chain sequence, or a portion thereof, is
derived from an anti-TREM2 antibody described herein (e.g., Clone
CL0020306, Clone CL0020188, or Clone CL0020307). The CDR, heavy
chain variable region, and light chain variable region amino acid
sequences of these clones is set forth in Table 8.
[0157] In some embodiments, an anti-TREM2 antibody comprises one or
more CDRs selected from the group consisting of:
[0158] (a) a heavy chain CDR1 (CDR-H1) sequence having at least 90%
sequence identity to the amino acid sequence of any one of SEQ ID
NOS:4 and 12, or having up to two amino acid substitutions relative
to the amino acid sequence of any one of SEQ ID NOS:4 and 12;
[0159] (b) a heavy chain CDR2 (CDR-H2) sequence having at least 90%
sequence identity to the amino acid sequence of any one of SEQ ID
NOS:5, 13, and 25, or having up to two amino acid substitutions
relative to the amino acid sequence of any one of SEQ ID NOS:5, 13,
and 25;
[0160] (c) a heavy chain CDR3 (CDR-H3) sequence having at least 90%
sequence identity to the amino acid sequence of any one of SEQ ID
NOS:6, 14, and 17, or having up to two amino acid substitutions
relative to the amino acid sequence of any one of SEQ ID NOS:6, 14,
and 17;
[0161] (d) a light chain CDR1 (CDR-L1) sequence having at least 90%
sequence identity to the amino acid sequence of any one of SEQ ID
NOS:7 and 23, or having up to two amino acid substitutions relative
to the amino acid sequence of any one of SEQ ID NOS:7 and 23;
[0162] (e) a light chain CDR2 (CDR-L2) sequence having at least 90%
sequence identity to the amino acid sequence of SEQ ID NO:8, or
having up to two amino acid substitutions relative to the amino
acid sequence of SEQ ID NO:8; and
[0163] (f) a light chain CDR3 (CDR-L3) sequence having at least 90%
sequence identity to the amino acid sequence of any one of SEQ ID
NOS:9 and 18, or having up to two amino acid substitutions relative
to the amino acid sequence of any one of SEQ ID NOS:9 and 18.
[0164] In some embodiments, an anti-TREM2 antibody comprises two,
three, four, five, or all six of (a)-(f). In some embodiments, an
anti-TREM2 antibody comprises the CDR-H1 of (a), the CDR-H2 of (b),
and the CDR-H3 of (c). In some embodiments, an anti-TREM2 antibody
comprises the CDR-L1 of (d), the CDR-L2 of (e), and the CDR-L3 of
(f). In some embodiments, a CDR having up to two amino acid
substitutions has one amino acid substitution relative to the
reference sequence. In some embodiments, a CDR having up to two
amino acid substitutions has two amino acid substitutions relative
to the reference sequence. In some embodiments, the up to two amino
acid substitutions are conservative substitutions.
[0165] In some embodiments, an anti-TREM2 antibody comprises one or
more CDRs selected from the group consisting of:
[0166] (a) a CDR-H1 sequence comprising the amino acid sequence of
any one of SEQ ID NOS:4 and 12;
[0167] (b) a CDR-H2 sequence comprising the amino acid sequence of
any one of SEQ ID NOS:5, 13, and 25;
[0168] (c) a CDR-H3 sequence comprising the amino acid sequence of
any one of SEQ ID NOS:6, 14, and 17;
[0169] (d) a CDR-L1 sequence comprising the amino acid sequence of
any one of SEQ ID NOS:7 and 23;
[0170] (e) a CDR-L2 sequence comprising the amino acid sequence of
SEQ ID NO:8; and
[0171] (f) a CDR-L3 sequence comprising the amino acid sequence of
any one of SEQ ID NOS:9 and 18.
[0172] In some embodiments, an anti-TREM2 antibody comprises two,
three, four, five, or all six of (a)-(f). In some embodiments, an
anti-TREM2 antibody comprises the CDR-H1 of (a), the CDR-H2 of (b),
and the CDR-H3 of (c). In some embodiments, an anti-TREM2 antibody
comprises the CDR-L1 of (d), the CDR-L2 of (e), and the CDR-L3 of
(f).
[0173] In some embodiments, an anti-TREM2 antibody comprises:
[0174] (a) a CDR-H1 comprising the amino acid sequence of SEQ ID
NO:4, a CDR-H2 comprising the amino acid sequence of SEQ ID NO:5, a
CDR-H3 comprising the amino acid sequence of SEQ ID NO:17, a CDR-L1
comprising the amino acid sequence of SEQ ID NO:7, a CDR-L2
comprising the amino acid sequence of SEQ ID NO:8, and a CDR-L3
comprising the amino acid sequence of SEQ ID NO:18; or
[0175] (b) a CDR-H1 comprising the amino acid sequence of SEQ ID
NO:4, a CDR-H2 comprising the amino acid sequence of SEQ ID NO:5, a
CDR-H3 comprising the amino acid sequence of SEQ ID NO:17, a CDR-L1
comprising the amino acid sequence of SEQ ID NO:23, a CDR-L2
comprising the amino acid sequence of SEQ ID NO:8, and a CDR-L3
comprising the amino acid sequence of SEQ ID NO:18; or
[0176] (c) a CDR-H1 comprising the amino acid sequence of SEQ ID
NO:4, a CDR-H2 comprising the amino acid sequence of SEQ ID NO:25,
a CDR-H3 comprising the amino acid sequence of SEQ ID NO:17, a
CDR-L1 comprising the amino acid sequence of SEQ ID NO:7, a CDR-L2
comprising the amino acid sequence of SEQ ID NO:8, and a CDR-L3
comprising the amino acid sequence of SEQ ID NO:18; or
[0177] (d) a CDR-H1 comprising the amino acid sequence of SEQ ID
NO:4, a CDR-H2 comprising the amino acid sequence of SEQ ID NO:25,
a CDR-H3 comprising the amino acid sequence of SEQ ID NO:17, a
CDR-L1 comprising the amino acid sequence of SEQ ID NO:23, a CDR-L2
comprising the amino acid sequence of SEQ ID NO:8, and a CDR-L3
comprising the amino acid sequence of SEQ ID NO:18; or
[0178] (e) a CDR-H1 comprising the amino acid sequence of SEQ ID
NO:4, a CDR-H2 comprising the amino acid sequence of SEQ ID NO:5, a
CDR-H3 comprising the amino acid sequence of SEQ ID NO:6, a CDR-L1
comprising the amino acid sequence of SEQ ID NO:7, a CDR-L2
comprising the amino acid sequence of SEQ ID NO:8, and a CDR-L3
comprising the amino acid sequence of SEQ ID NO:9; or
[0179] (f) a CDR-H1 comprising the amino acid sequence of SEQ ID
NO:12, a CDR-H2 comprising the amino acid sequence of SEQ ID NO:13,
a CDR-H3 comprising the amino acid sequence of SEQ ID NO:14, a
CDR-L1 comprising the amino acid sequence of SEQ ID NO:7, a CDR-L2
comprising the amino acid sequence of SEQ ID NO:8, and a CDR-L3
comprising the amino acid sequence of SEQ ID NO:9; or
[0180] (g) a CDR-H1 comprising the amino acid sequence of SEQ ID
NO:4, a CDR-H2 comprising the amino acid sequence of SEQ ID NO:25,
a CDR-H3 comprising the amino acid sequence of SEQ ID NO:17, a
CDR-L1 comprising the amino acid sequence of SEQ ID NO:7, a CDR-L2
comprising the amino acid sequence of SEQ ID NO:8, and a CDR-L3
comprising the amino acid sequence of SEQ ID NO:9.
[0181] In some embodiments, an anti-TREM2 antibody comprises a
heavy chain variable region comprising an amino acid sequence that
has at least 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or
99% sequence identity to any one of SEQ ID NOS:2, 10, 15, 19, 21,
24, and 26. In some embodiments, an anti-TREM2 comprises a heavy
chain variable region comprising the amino acid sequence of any one
of SEQ ID NOS:2, 10, 15, 19, 21, 24, and 26.
[0182] In some embodiments, an anti-TREM2 antibody comprises a
light chain variable region comprising an amino acid sequence that
has at least 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or
99% sequence identity to any one of SEQ ID NOS:3, 11, 16, 20, 22,
and 27. In some embodiments, an anti-TREM2 antibody comprises a
light chain variable region comprising the amino acid sequence of
any one of SEQ ID NOS:3, 11, 16, 20, 22, and 27.
[0183] In some embodiments, an anti-TREM2 antibody comprises: a
heavy chain variable region comprising an amino acid sequence that
has at least 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or
99% sequence identity to any one of SEQ ID NOS:2, 10, 15, 19, 21,
24, and 26, a light chain variable region comprising an amino acid
sequence that has at least 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%,
97%, 98%, or 99% sequence identity to any one of SEQ ID NOS:3, 11,
16, 20, 22, and 27. In some embodiments, an anti-TREM2 comprises: a
heavy chain variable region comprising the amino acid sequence of
any one of SEQ ID NOS:2, 10, 15, 19, 21, 24, and 26, and a light
chain variable region comprising the amino acid sequence of any one
of SEQ ID NOS:3, 11, 16, 20, 22, and 27.
[0184] In some embodiments, an anti-TREM2 antibody comprises:
[0185] (a) a V.sub.H sequence that has at least 85% sequence
identity to SEQ ID NO:2 and a V.sub.L sequence has at least 85%
sequence identity to SEQ ID NO:3; or
[0186] (b) a V.sub.H sequence that has at least 85% sequence
identity to SEQ ID NO:10 and a V.sub.L sequence has at least 85%
sequence identity to SEQ ID NO:11; or
[0187] (c) a V.sub.H sequence that has at least 85% sequence
identity to SEQ ID NO:15 and a V.sub.L sequence has at least 85%
sequence identity to SEQ ID NO:16; or
[0188] (d) a V.sub.H sequence that has at least 85% sequence
identity to SEQ ID NO:19 and a V.sub.L sequence has at least 85%
sequence identity to SEQ ID NO:20; or
[0189] (e) a V.sub.H sequence that has at least 85% sequence
identity to SEQ ID NO:21 and a V.sub.L sequence has at least 85%
sequence identity to SEQ ID NO:20; or
[0190] (f) a V.sub.H sequence that has at least 85% sequence
identity to SEQ ID NO:19 and a V.sub.L sequence has at least 85%
sequence identity to SEQ ID NO:22; or
[0191] (g) a V.sub.H sequence that has at least 85% sequence
identity to SEQ ID NO:21 and a V.sub.L sequence has at least 85%
sequence identity to SEQ ID NO:22; or
[0192] (h) a V.sub.H sequence that has at least 85% sequence
identity to SEQ ID NO:24 and a V.sub.L sequence has at least 85%
sequence identity to SEQ ID NO:20; or
[0193] (i) a V.sub.H sequence that has at least 85% sequence
identity to SEQ ID NO:26 and a V.sub.L sequence has at least 85%
sequence identity to SEQ ID NO:20; or
[0194] (j) a V.sub.H sequence that has at least 85% sequence
identity to SEQ ID NO:24 and a V.sub.L sequence has at least 85%
sequence identity to SEQ ID NO:22; or
[0195] (k) a V.sub.H sequence that has at least 85% sequence
identity to SEQ ID NO:26 and a V.sub.L sequence has at least 85%
sequence identity to SEQ ID NO:22; or
[0196] (l) a V.sub.H sequence that has at least 85% sequence
identity to SEQ ID NO:24 and a V.sub.L sequence has at least 85%
sequence identity to SEQ ID NO:27.
[0197] In some embodiments, an anti-TREM2 antibody comprises one or
more sequences that are encompassed by a consensus sequence
disclosed herein. As a non-limiting example, consensus sequences
can be identified by aligning heavy chain or light chain sequences
(e.g., CDRs) for antibodies that are from the same (or similar)
germlines. In some embodiments, consensus sequences may be
generated from antibodies that contain sequences that are of the
same (or similar) length and/or have at least one highly similar
CDR (e.g., a highly similar CDR3). In some embodiments, such
sequences in these antibodies may be aligned and compared to
identify conserved amino acids or motifs (i.e., where alteration in
sequences may alter protein function) and/or regions where
variation occurs the sequences (i.e., where variation of sequence
is not likely to significantly affect protein function).
Alternatively, consensus sequences can be identified by aligning
heavy chain or light chain sequences (e.g., CDRs) for antibodies
that bind to the same or similar (e.g., overlapping) epitopes to
determine conserved amino acids or motifs (i.e., where alteration
in sequences may alter protein function) and regions where
variation occurs in alignment of sequences (i.e., where variation
of sequence is not likely to significantly affect protein
function). In some embodiments, one or more consensus sequences can
be identified for antibodies that recognize the same or similar
epitope as an anti-TREM2 antibody as disclosed herein. Exemplary
consensus sequences include SEQ ID NOS:28-32. In the consensus
sequences of SEQ ID NOS:28-32, the capitalized letter represents an
amino acid residue that is absolutely conserved among the aligned
sequences (e.g., aligned CDR sequences), while an "X" or a Greek
letter (e.g., ".alpha.," ".beta.," ".gamma.," ".delta.,"
".epsilon.," or ".phi.") represents an amino acid residue that is
not absolutely conserved among the aligned sequences. It will be
appreciated that, when selecting an amino acid to insert at a
position marked by an "X" or by a Greek letter, in some embodiments
the amino acid is selected from those amino acids found at the
corresponding position in the aligned sequences.
[0198] Clones CL0020188, CL0020306, CL0020307, and Variants of
CL0020188
[0199] In some embodiments, an anti-TREM2 antibody comprises:
[0200] (a) a CDR-H1 sequence comprising the sequence of
G-F-T-F-T-.alpha..sub.6-F-Y-M-S (SEQ ID NO:28), wherein
.alpha..sub.6 is D or N;
[0201] (b) a CDR-H2 sequence comprising the sequence of
V-I-R-N-.beta..sub.5-.beta..sub.6-N-.beta..sub.8-Y-T-.beta..sub.11-.beta.-
.sub.12-Y-N-P-S-V-K-G (SEQ ID NO:29), wherein .beta..sub.5 is K or
R; .beta..sub.6 is A or P; .beta..sub.8 is G or A; .beta..sub.11 is
A or T; and .beta..sub.12 is G or D;
[0202] (c) a CDR-H3 sequence comprising the sequence of
.gamma..sub.1-R-L-.gamma..sub.4-Y-G-F-D-Y (SEQ ID NO:30), wherein
.gamma..sub.1 is A or T; and .gamma..sub.4 is T or S;
[0203] (d) a CDR-L1 sequence comprising the sequence of
Q-S-S-K-S-L-L-H-S-.delta..sub.10-G-K-T-Y-L-N(SEQ ID NO:31), wherein
.delta..sub.10 is N or T;
[0204] (e) a CDR-L2 sequence comprising the sequence of WMSTRAS
(SEQ ID NO:8); and
[0205] (f) a CDR-L3 sequence comprising the sequence of
Q-Q-F-L-E-.PHI..sub.6-P-F-T (SEQ ID NO:32), wherein .PHI..sub.6 is
Y or F.
[0206] In some embodiments, an anti-TREM2 antibody comprises a
CDR-H1 sequence that is selected from SEQ ID NOS:4 and 12. In some
embodiments, an anti-TREM2 antibody comprises a CDR-H2 sequence
that is selected from SEQ ID NOS:5, 13, and 25. In some
embodiments, an anti-TREM2 antibody comprises a CDR-H3 sequence
that is selected from SEQ ID NOS:6, 14, and 17. In some
embodiments, an anti-TREM2 antibody comprises a CDR-L1 sequence
that is selected from SEQ ID NOS:7 and 23. In some embodiments, an
anti-TREM2 antibody comprises a CDR-L3 sequence is selected from
SEQ ID NOS:9 and 18.
[0207] In some embodiments, an anti-TREM2 antibody comprises a
heavy chain variable region comprising an amino acid sequence that
has at least 85% sequence identity (e.g., at least 90%, 91%, 92%,
93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to any one
of SEQ ID NOS:2, 10, 15, 19, 21, 24, and 26. In some embodiments,
an anti-TREM2 antibody comprises a heavy chain variable region
comprising the amino acid sequence of any one of SEQ ID NOS:2, 10,
15, 19, 21, 24, and 26.
[0208] In some embodiments, an anti-TREM2 antibody comprises a
light chain variable region comprising an amino acid sequence that
has at least 85% sequence identity (e.g., at least 90%, 91%, 92%,
93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to any one
of SEQ ID NOS:3, 11, 16, 20, 22, and 27. In some embodiments, an
anti-TREM2 antibody comprises a light chain variable region
comprising the amino acid sequence of any one of SEQ ID NOS:3, 11,
16, 20, 22, and 27.
Clone CL0020188
[0209] In some embodiments, an anti-TREM2 antibody comprises a
CDR-H1 sequence comprising the amino acid sequence of SEQ ID NO:4,
a CDR-H2 sequence comprising the amino acid sequence of SEQ ID
NO:5, a CDR-H3 sequence comprising the amino acid sequence of SEQ
ID NO:17, a CDR-L1 sequence comprising the amino acid sequence of
SEQ ID NO:7, a CDR-L2 sequence comprising the amino acid sequence
of SEQ ID NO:8, and a CDR-L3 sequence comprising the amino acid
sequence of SEQ ID NO:18.
[0210] In some embodiments, an anti-TREM2 antibody comprises a
heavy chain variable region comprising an amino acid sequence that
has at least 85% sequence identity (e.g., at least 90%, 91%, 92%,
93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to SEQ ID
NO:15. In some embodiments, an anti-TREM2 antibody comprises a
heavy chain variable region comprising the amino acid sequence of
SEQ ID NO:15.
[0211] In some embodiments, an anti-TREM2 antibody comprises a
light chain variable region comprising an amino acid sequence that
has at least 85% sequence identity (e.g., at least 90%, 91%, 92%,
93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to SEQ ID
NO:16. In some embodiments, an anti-TREM2 antibody comprises a
light chain variable region comprising the amino acid sequence of
SEQ ID NO:16.
[0212] In some embodiments, an anti-TREM2 antibody comprises a
heavy chain variable region comprising an amino acid sequence that
has at least 85% sequence identity (e.g., at least 90%, 91%, 92%,
93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to SEQ ID
NO:15 and a light chain variable region comprising an amino acid
sequence that has at least 85% sequence identity (e.g., at least
90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence
identity) to SEQ ID NO:16. In some embodiments, an anti-TREM2
antibody comprises a heavy chain variable region comprising the
amino acid sequence of SEQ ID NO:15 and a light chain variable
region comprising the amino acid sequence of SEQ ID NO: 16.
[0213] In some embodiments, an anti-TREM2 antibody comprises a
heavy chain variable region that comprises a heavy chain CDR1-3
comprising the amino acid sequences of SEQ ID NOS:4, 5, and 17,
respectively, and that has at least 85% sequence identity (e.g., at
least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence
identity) to SEQ ID NO:15. In some embodiments, an anti-TREM2
antibody comprises a light chain variable region that comprises a
light chain CDR1-3 comprising the amino acid sequences of SEQ ID
NOS:7, 8, and 18, respectively, and that has at least 85% sequence
identity (e.g., at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%,
98%, or 99% sequence identity) to SEQ ID NO:16.
[0214] In some embodiments, an anti-TREM2 antibody competes for
binding with an antibody as described herein (e.g., an antibody
comprising a heavy chain CDR1-3 and a light chain CDR1-3 comprising
the amino acid sequences of SEQ ID NOS:4, 5, 17, 7, 8, and 18,
respectively, or an antibody comprising a heavy chain variable
region comprising the amino acid sequence of SEQ ID NO:15 and a
light chain variable region comprising the amino acid sequence of
SEQ ID NO:16).
[0215] In some embodiments, an anti-TREM2 antibody comprises a
CDR-H1 sequence comprising the amino acid sequence of SEQ ID NO:4,
a CDR-H2 sequence comprising the amino acid sequence of SEQ ID
NO:25, a CDR-H3 sequence comprising the amino acid sequence of SEQ
ID NO:17, a CDR-L1 sequence comprising the amino acid sequence of
SEQ ID NO:23, a CDR-L2 sequence comprising the amino acid sequence
of SEQ ID NO:8, and a CDR-L3 sequence comprising the amino acid
sequence of SEQ ID NO:18.
[0216] In some embodiments, an anti-TREM2 antibody or antigen
binding portion comprises a heavy chain variable region comprising
an amino acid sequence that has at least 85% sequence identity
(e.g., at least 90%, 95%, or 97% sequence identity) to SEQ ID
NO:24. In some embodiments, an anti-TREM2 antibody or antigen
binding portion comprises a heavy chain variable region comprising
the amino acid sequence of SEQ ID NO:24.
[0217] In some embodiments, an anti-TREM2 antibody or antigen
binding portion comprises a light chain variable region comprising
an amino acid sequence that has at least 85% sequence identity
(e.g., at least 90%, 95%, or 97% sequence identity) to SEQ ID
NO:22. In some embodiments, an anti-TREM2 antibody or antigen
binding portion comprises a light chain variable region comprising
the amino acid sequence of SEQ ID NO:22.
[0218] In some embodiments, an anti-TREM2 antibody or antigen
binding portion comprises a heavy chain variable region comprising
an amino acid sequence that has at least 85% sequence identity
(e.g., at least 90%, 95%, or 97% sequence identity) to SEQ ID NO:24
and a light chain variable region comprising an amino acid sequence
that has at least 85% sequence identity (e.g., at least 90%, 95%,
or 97% sequence identity) to SEQ ID NO:22. In some embodiments, an
anti-TREM2 antibody or antigen binding portion comprises a heavy
chain variable region comprising the amino acid sequence of SEQ ID
NO:24 and a light chain variable region comprising the amino acid
sequence of SEQ ID NO:22.
[0219] In some embodiments, an anti-TREM2 antibody or antigen
binding portion comprises a heavy chain variable region that
comprises a heavy chain CDR1-3 comprising the amino acid sequences
of SEQ ID NOS:4, 25, and 17, respectively, and that has at least
85% sequence identity (e.g., at least 90%, 95%, or 97% sequence
identity) to SEQ ID NO:24. In some embodiments, an anti-TREM2
antibody or antigen binding portion comprises a light chain
variable region that comprises a light chain CDR1-3 comprising the
amino acid sequences of SEQ ID NOS:23, 8, and 18, respectively, and
that has at least 85% sequence identity (e.g., at least 90%, 95%,
or 97% sequence identity) to SEQ ID NO:22.
[0220] In some embodiments, an anti-TREM2 antibody or antigen
binding portion competes for binding with an antibody as described
herein (e.g., an antibody comprising a heavy chain CDR1-3 and a
light chain CDR1-3 comprising the amino acid sequences of SEQ ID
NOS:4, 25, 17, 23, 8, and 18, respectively, or an antibody
comprising a heavy chain variable region comprising the amino acid
sequence of SEQ ID NO:24 and a light chain variable region
comprising the amino acid sequence of SEQ ID NO:22).
[0221] In some embodiments, an anti-TREM2 antibody or antigen
binding portion comprises a CDR-H1 sequence comprising the amino
acid sequence of SEQ ID NO:4, a CDR-H2 sequence comprising the
amino acid sequence of SEQ ID NO:25, a CDR-H3 sequence comprising
the amino acid sequence of SEQ ID NO:17, a CDR-L1 sequence
comprising the amino acid sequence of SEQ ID NO:7, a CDR-L2
sequence comprising the amino acid sequence of SEQ ID NO:8, and a
CDR-L3 sequence comprising the amino acid sequence of SEQ ID
NO:9.
[0222] In some embodiments, an anti-TREM2 antibody or antigen
binding portion comprises a heavy chain variable region comprising
an amino acid sequence that has at least 85% sequence identity
(e.g., at least 90%, 95%, or 97% sequence identity) to SEQ ID
NO:24. In some embodiments, an anti-TREM2 antibody or antigen
binding portion comprises a heavy chain variable region comprising
the amino acid sequence of SEQ ID NO:24.
[0223] In some embodiments, an anti-TREM2 antibody or antigen
binding portion comprises a light chain variable region comprising
an amino acid sequence that has at least 85% sequence identity
(e.g., at least 90%, 95%, or 97% sequence identity) to SEQ ID
NO:27. In some embodiments, an anti-TREM2 antibody or antigen
binding portion comprises a light chain variable region comprising
the amino acid sequence of SEQ ID NO:27.
[0224] In some embodiments, an anti-TREM2 antibody or antigen
binding portion comprises a heavy chain variable region comprising
an amino acid sequence that has at least 85% sequence identity
(e.g., at least 90%, 95%, or 97% sequence identity) to SEQ ID NO:24
and a light chain variable region comprising an amino acid sequence
that has at least 85% sequence identity (e.g., at least 90%, 95%,
or 97% sequence identity) to SEQ ID NO:27. In some embodiments, an
anti-TREM2 antibody or antigen binding portion comprises a heavy
chain variable region comprising the amino acid sequence of SEQ ID
NO:24 and a light chain variable region comprising the amino acid
sequence of SEQ ID NO:27.
[0225] In some embodiments, an anti-TREM2 antibody or antigen
binding portion comprises a heavy chain variable region that
comprises a heavy chain CDR1-3 comprising the amino acid sequences
of SEQ ID NOS:4, 25, and 17, respectively, and that has at least
85% sequence identity (e.g., at least 90%, 95%, or 97% sequence
identity) to SEQ ID NO:24. In some embodiments, an anti-TREM2
antibody or antigen binding portion comprises a light chain
variable region that comprises a light chain CDR1-3 comprising the
amino acid sequences of SEQ ID NOS:7, 8, and 9, respectively, and
that has at least 85% sequence identity (e.g., at least 90%, 95%,
or 97% sequence identity) to SEQ ID NO:27.
[0226] In some embodiments, an anti-TREM2 antibody or antigen
binding portion is an antibody that competes for binding with an
antibody as described herein (e.g., an antibody comprising a heavy
chain CDR1-3 and a light chain CDR1-3 comprising the amino acid
sequences of SEQ ID NOS:4, 25, 17, 7, 8, and 9, respectively, or an
antibody comprising a heavy chain variable region comprising the
amino acid sequence of SEQ ID NO:24 and a light chain variable
region comprising the amino acid sequence of SEQ ID NO:27).
Clone CL0020306
[0227] In some embodiments, an anti-TREM2 antibody comprises a
CDR-H1 sequence comprising the amino acid sequence of SEQ ID NO:4,
a CDR-H2 sequence comprising the amino acid sequence of SEQ ID
NO:5, a CDR-H3 sequence comprising the amino acid sequence of SEQ
ID NO:6, a CDR-L1 sequence comprising the amino acid sequence of
SEQ ID NO:7, a CDR-L2 sequence comprising the amino acid sequence
of SEQ ID NO:8, and a CDR-L3 sequence comprising the amino acid
sequence of SEQ ID NO:9.
[0228] In some embodiments, an anti-TREM2 antibody comprises a
heavy chain variable region comprising an amino acid sequence that
has at least 85% sequence identity (e.g., at least 90%, 91%, 92%,
93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to SEQ ID
NO:2. In some embodiments, an anti-TREM2 antibody comprises a heavy
chain variable region comprising the amino acid sequence of SEQ ID
NO:2.
[0229] In some embodiments, an anti-TREM2 antibody comprises a
light chain variable region comprising an amino acid sequence that
has at least 85% sequence identity (e.g., at least 90%, 91%, 92%,
93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to SEQ ID
NO:3. In some embodiments, an anti-TREM2 antibody comprises a light
chain variable region comprising the amino acid sequence of SEQ ID
NO:3.
[0230] In some embodiments, an anti-TREM2 antibody comprises a
heavy chain variable region comprising an amino acid sequence that
has at least 85% sequence identity (e.g., at least 90%, 91%, 92%,
93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to SEQ ID
NO:2 and a light chain variable region comprising an amino acid
sequence that has at least 85% sequence identity (e.g., at least
90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence
identity) to SEQ ID NO:3. In some embodiments, an anti-TREM2
antibody comprises a heavy chain variable region comprising the
amino acid sequence of SEQ ID NO:2 and a light chain variable
region comprising the amino acid sequence of SEQ ID NO:3.
[0231] In some embodiments, an anti-TREM2 antibody comprises a
heavy chain variable region that comprises a heavy chain CDR1-3
comprising the amino acid sequences of SEQ ID NOS:4, 5, and 6,
respectively, and that has at least 85% sequence identity (e.g., at
least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence
identity) to SEQ ID NO:2. In some embodiments, an anti-TREM2
antibody comprises a light chain variable region that comprises a
light chain CDR1-3 comprising the amino acid sequences of SEQ ID
NOS:7, 8, and 9, respectively, and that has at least 85% sequence
identity (e.g., at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%,
98%, or 99% sequence identity) to SEQ ID NO:3.
[0232] In some embodiments, an anti-TREM2 antibody competes for
binding with an antibody as described herein (e.g., an antibody
comprising a heavy chain CDR1-3 and a light chain CDR1-3 comprising
the amino acid sequences of SEQ ID NOS:4, 5, 6, 7, 8, and 9,
respectively, or an antibody comprising a heavy chain variable
region comprising the amino acid sequence of SEQ ID NO:2 and a
light chain variable region comprising the amino acid sequence of
SEQ ID NO:3).
Clone CL0020307
[0233] In some embodiments, an anti-TREM2 antibody comprises a
CDR-H1 sequence comprising the amino acid sequence of SEQ ID NO:12,
a CDR-H2 sequence comprising the amino acid sequence of SEQ ID
NO:13, a CDR-H3 sequence comprising the amino acid sequence of SEQ
ID NO:14, a CDR-L1 sequence comprising the amino acid sequence of
SEQ ID NO:7, a CDR-L2 sequence comprising the amino acid sequence
of SEQ ID NO:8, and a CDR-L3 sequence comprising the amino acid
sequence of SEQ ID NO:9.
[0234] In some embodiments, an anti-TREM2 antibody comprises a
heavy chain variable region comprising an amino acid sequence that
has at least 85% sequence identity (e.g., at least 90%, 91%, 92%,
93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to SEQ ID
NO:10. In some embodiments, an anti-TREM2 antibody or antigen
binding portion comprises a heavy chain variable region comprising
the amino acid sequence of SEQ ID NO:10.
[0235] In some embodiments, an anti-TREM2 antibody comprises a
light chain variable region comprising an amino acid sequence that
has at least 85% sequence identity (e.g., at least 90%, 91%, 92%,
93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to SEQ ID
NO:11. In some embodiments, an anti-TREM2 antibody comprises a
light chain variable region comprising the amino acid sequence of
SEQ ID NO:11.
[0236] In some embodiments, an anti-TREM2 antibody comprises a
heavy chain variable region comprising an amino acid sequence that
has at least 85% sequence identity (e.g., at least 90%, 91%, 92%,
93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to SEQ ID
NO:10 and a light chain variable region comprising an amino acid
sequence that has at least 85% sequence identity (e.g., at least
90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence
identity) to SEQ ID NO:11. In some embodiments, an anti-TREM2
antibody comprises a heavy chain variable region comprising the
amino acid sequence of SEQ ID NO:10 and a light chain variable
region comprising the amino acid sequence of SEQ ID NO: 11.
[0237] In some embodiments, an anti-TREM2 antibody comprises a
heavy chain variable region that comprises a heavy chain CDR1-3
comprising the amino acid sequences of SEQ ID NOS:12, 13, and 14,
respectively, and that has at least 85% sequence identity (e.g., at
least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence
identity) to SEQ ID NO:10. In some embodiments, an anti-TREM2
antibody comprises a light chain variable region that comprises a
light chain CDR1-3 comprising the amino acid sequences of SEQ ID
NOS:7, 8, and 9, respectively, and that has at least 85% sequence
identity (e.g., at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%,
98%, or 99% sequence identity) to SEQ ID NO:11.
[0238] In some embodiments, an anti-TREM2 antibody is an antibody
that competes for binding with an antibody as described herein
(e.g., an antibody comprising a heavy chain CDR1-3 and a light
chain CDR1-3 comprising the amino acid sequences of SEQ ID NOS:12,
13, 14, 7, 8, and 9, respectively, or an antibody comprising a
heavy chain variable region comprising the amino acid sequence of
SEQ ID NO:10 and a light chain variable region comprising the amino
acid sequence of SEQ ID NO: 11).
Binding Characteristics of Anti-TREM2 Antibodies
[0239] In some embodiments, an antibody as described herein that
specifically binds to a TREM2 protein binds to TREM2 that is
expressed on a cell (e.g., a primary cell or cell line that
endogenously expresses TREM2, such as human macrophages, or a
primary cell or cell line that has been engineered to express
TREM2, e.g., as described in the Examples section below). In some
embodiments, an antibody that specifically binds to a TREM2 protein
as described herein binds to purified or recombinant TREM2 protein
of a portion thereof, or to a chimeric protein comprising TREM2 or
a portion thereof (e.g., an Fc-fusion protein comprising TREM2 or
an Fc-fusion protein comprising the ecto-domain of TREM2).
[0240] In some embodiments, some embodiments, an antibody that
specifically binds to human TREM2 protein exhibits cross-reactivity
with one or more other TREM2 proteins of another species. In some
embodiments, an antibody that specifically binds to human TREM2
protein exhibits cross-reactivity with a cynomolgus monkey ("cyno")
TREM2 protein. In some embodiments, an antibody that specifically
binds to human TREM2 protein exhibits cross-reactivity with a mouse
TREM2 protein. In some embodiments, an anti-TREM2 antibody exhibits
cross-reactivity with human TREM2, cyno TREM2, and mouse TREM2.
[0241] Methods for analyzing binding affinity, binding kinetics,
and cross-reactivity are known in the art. These methods include,
but are not limited to, solid-phase binding assays (e.g., ELISA
assay), immunoprecipitation, surface plasmon resonance (e.g.,
Biacore.TM. (GE Healthcare, Piscataway, N.J.)), kinetic exclusion
assays (e.g., KinExA.COPYRGT.), flow cytometry,
fluorescence-activated cell sorting (FACS), BioLayer interferometry
(e.g., Octet.TM. (ForteBio, Inc., Menlo Park, Calif.)), and western
blot analysis. In some embodiments, ELISA is used to determine
binding affinity and/or cross-reactivity. Methods for performing
ELISA assays are known in the art, and are also described in the
Examples section below. In some embodiments, surface plasmon
resonance (SPR) is used to determine binding affinity, binding
kinetics, and/or cross-reactivity. In some embodiments, kinetic
exclusion assays are used to determine binding affinity, binding
kinetics, and/or cross-reactivity. In some embodiments, BioLayer
interferometry assays are used to determine binding affinity,
binding kinetics, and/or cross-reactivity.
Epitopes Recognized by Anti-TREM2 Antibodies
[0242] In some embodiments, an anti-TREM2 antibody recognizes an
epitope of human TREM2 that is the same or substantially the same
as the epitope recognized by an antibody clone as described herein.
As used herein, the term "substantially the same," as used with
reference to an epitope recognized by an antibody clone as
described herein, means that the anti-TREM2 antibody recognizes an
epitope that is identical, within, or nearly identical to (e.g.,
has at least 90% sequence identity to, or has one, two, or three
amino acid substitutions, e.g., conservative substitutions,
relative to), or has substantial overlap with (e.g., at least 50%,
60%, 70%, 80%, 90%, or 95% overlap with) the epitope recognized by
the antibody clone as described herein.
[0243] In some embodiments, an anti-TREM2 antibody recognizes an
epitope of human TREM2 that is the same or substantially the same
as the epitope recognized by an antibody clone selected from the
group consisting of Clone CL0020306, Clone CL0020188, Clone
CL0020307, and variants of the same.
[0244] In some embodiments, an anti-TREM2 antibody binds to human
TREM2 at an epitope within the stalk region of TREM2. In some
embodiments, an anti-TREM2 antibody recognizes an epitope of human
TREM2 comprising, within, or consisting of residues 129-172 or
residues 131-169 of SEQ ID NO:1. In some embodiments, an anti-TREM2
antibody recognizes an epitope of human TREM2 comprising, within,
or consisting of residues 129-148 of SEQ ID NO:1 (e.g., 143-148 of
SEQ ID NO:1). In some embodiments, an anti-TREM2 antibody is an
agonist that activates TREM2/DAP12 signaling (e.g., by inducing
phosphorylation of a kinase such as Syk) and binds to human TREM2
at an epitope within the stalk region of TREM2. In some
embodiments, an anti-TREM2 antibody binds to human TREM2 at an
epitope within the stalk region of TREM2 and inhibits cleavage of
TREM2 by a protease (e.g., ADAM17).
Functional Characteristics of Anti-TREM2 Antibodies
[0245] In some embodiments, an anti-TREM2 antibody (e.g., an
antibody having one or more CDR, heavy chain variable region,
and/or light chain variable region sequences as disclosed)
functions in one or more TREM2 activities as disclosed herein. For
example, in some embodiments an anti-TREM2 antibody modulates
levels of sTREM2 protein (e.g., levels of sTREM2 that are shed from
the cell surface into an extracellular sample), modulates
recruitment or phosphorylation of a kinase that interacts with a
TREM2/DAP12 signaling complex (e.g., Syk kinase), and/or modulates
one or more activities downstream of the signaling complex, such as
phagocytosis, cell growth, cell survival, cell differentiation,
cytokine secretion, or cell migration.
[0246] In some embodiments, an anti-TREM2 antibody enhances one or
more TREM2 activities (e.g., those described herein) that are
induced by a ligand. In some embodiments, the ligand is a lipid
ligand. Examples of TREM2 lipid ligands include, but are not
limited to,
1-palmitoyl-2-(5'-oxo-valeroyl)-sn-glycero-3-phosphocholine
(POVPC), 2-Arachidonoylglycerol (2-AG), 7-ketocholesterol (7-KC),
24(S)hydroxycholesterol (24OHC), 25(S)hydroxycholesterol (25OHC),
27-hydroxycholesterol (27OHC), Acyl Carnitine (AC),
alkylacylglycerophosphocholine (PAF), .alpha.-galactosylceramide
(KRN7000), Bis(monoacylglycero)phosphate (BMP), Cardiolipin (CL),
Ceramide, Ceramide-1-phosphate (C1P), Cholesteryl ester (CE),
Cholesterol phosphate (CP), Diacylglycerol 34:1 (DG 34:1),
Diacylglycerol 38:4 (DG 38:4), Diacylglycerol pyrophosphate (DGPP),
Dihyrdoceramide (DhCer), Dihydrosphingomyelin (DhSM), Ether
phosphatidylcholine (PCe), Free cholesterol (FC),
Galactosylceramide (GalCer), Galactosylsphingosine (GalSo),
Ganglioside GM1, Ganglioside GM3, Glucosylsphingosine (GlcSo),
Hank's Balanced Salt Solution (HBSS), Kdo2-Lipid A (KLA),
Lactosylceramide (LacCer), lysoalkylacylglycerophosphocholine
(LPAF), Lysophosphatidic acid (LPA), Lysophosphatidylcholine (LPC),
Lysophosphatidylethanolamine (LPE), Lysophosphatidylglycerol (LPG),
Lysophosphatidylinositol (LPI), Lysosphingomyelin (LSM),
Lysophosphatidylserine (LPS), N-Acyl-phosphatidylethanolamine
(NAPE), N-Acyl-Serine (NSer), Oxidized phosphatidylcholine (oxPC),
Palmitic-acid-9-hydroxy-stearic-acid (PAHSA),
Phosphatidylethanolamine (PE), Phosphatidylethanol (PEtOH),
Phosphatidic acid (PA), Phosphatidylcholine (PC),
Phosphatidylglycerol (PG), Phosphatidylinositol (PI),
Phosphatidylserine (PS), Sphinganine, Sphinganine-1-phosphate
(SalP), Sphingomyelin (SM), Sphingosine, Sphingosine-1-phosphate
(So1P), and Sulfatide.
[0247] Modulation of sTREM2 Shedding
[0248] In some embodiments, an anti-TREM2 antibody alters levels of
sTREM2 protein in a sample, e.g., levels of sTREM2 that are shed
from the cell surface into an extracellular sample. In some
embodiments, an anti-TREM2 antibody decreases levels of sTREM2.
[0249] In some embodiments, an anti-TREM2 antibody decreases levels
of sTREM2 if the amount of sTREM2 in a treated sample is decreased
by at least 10%, at least 20%, at least 30%, at least 40%, at least
50%, at least 60%, at least 70%, at least 80%, at least 90% or more
as compared to a control value. In some embodiments, an anti-TREM2
antibody decreases levels of sTREM2 if the amount of sTREM2 in a
treated sample is decreased by at least 2-fold, 3-fold, 4-fold,
5-fold, 6-fold, 7-fold, 8-fold, 9-fold, 10-fold or more as compared
to a control value. In some embodiments, the control value is the
amount of sTREM2 in an untreated sample (e.g., a supernatant from a
TREM2-expressing cell that has not been treated with an anti-TREM2
antibody, or a sample from a subject that has not been treated with
an anti-TREM2 antibody) or a sample treated with an appropriate
non-TREM2-binding antibody.
[0250] In some embodiments, sTREM2 shedding is measured using a
sample that comprises a fluid, e.g., blood, plasma, serum, urine,
or cerebrospinal fluid. In some embodiments, the sample comprises
cerebrospinal fluid. In some embodiments, the sample comprises
supernatant from cell cultures (e.g., supernatant from a primary
cell or cell line that endogenously expresses TREM2, such as human
macrophages, or a primary cell or cell line that has been
engineered to express TREM2, e.g., as described in the Examples
section below).
[0251] In some embodiments, the level of sTREM2 in a sample is
measured using an immunoassay. Immunoassays are known in the art
and include, but are not limited to, enzyme immunoassays (EIA) such
as enzyme multiplied immunoassay (EMIA), enzyme-linked
immunosorbent assay (ELISA), microparticle enzyme immunoassay
(MEIA), immunohistochemistry (IHC), immunocytochemistry, capillary
electrophoresis immunoassays (CEIA), radioimmunoassays (RIA),
immunofluorescence, chemiluminescence immunoassays (CL), and
electrochemiluminescence immunoassays (ECL). In some embodiments,
sTREM2 levels are measuring using an ELISA assay. In some
embodiments, sTREM2 levels are measured using an ELISA assay as
described in the Examples section below.
[0252] Modulation of Kinase Recruitment or Phosphorylation
[0253] In some embodiments, an anti-TREM2 antibody induces
phosphorylation of a kinase that interacts with the TREM2/DAP12
signaling complex (such as, but not limited to, Syk, ZAP70, PI3K,
Erk, AKT, or GSK3b). In some embodiments, an anti-TREM2 antibody
induces phosphorylation of a kinase that interacts with the
TREM2/DAP12 signaling complex without blocking binding of a native
TREM2 ligand. In some embodiments, an anti-TREM2 antibody enhances
phosphorylation of a kinase that interacts with the TREM2/DAP12
signaling complex that is induced by a TREM2 ligand (e.g., a lipid
ligand). In some embodiments, an anti-TREM2 antibody induces or
enhances phosphorylation of Syk. In some embodiments, an anti-TREM2
antibody induces or enhances phosphorylation of Syk if the level of
Syk phosphorylation in a sample treated with the anti-TREM2
antibody is increased by at least 10%, at least 20%, at least 30%,
at least 40%, at least 50%, at least 60%, at least 70%, at least
80%, at least 90% or more as compared to a control value. In some
embodiments, an anti-TREM2 antibody induces phosphorylation of Syk
if the level of Syk phosphorylation in a sample treated with the
anti-TREM2 antibody is increased by at least 2-fold, 3-fold,
4-fold, 5-fold, 6-fold, 7-fold, 8-fold, 9-fold, 10-fold, or more as
compared to a control value. In some embodiments, the control value
is the level of Syk phosphorylation in an untreated sample (e.g., a
sample comprising a TREM2-expressing cell that has not been treated
with an anti-TREM2 antibody, or a sample from a subject that has
not been treated with an anti-TREM2 antibody), or a sample that has
been treated with a TREM2 ligand but not an anti-TREM2 antibody, or
a sample treated with an appropriate non-TREM2-binding
antibody.
[0254] For detecting and/or quantifying phosphorylation (e.g., Syk
phosphorylation) in a sample, in some embodiments, an immunoassay
is used. In some embodiments, the immunoassay is an enzyme
immunoassay (EIA), enzyme multiplied immunoassay (EMIA),
enzyme-linked immunosorbent assay (ELISA), microparticle enzyme
immunoassay (MEIA), immunohistochemistry (IHC),
immunocytochemistry, capillary electrophoresis immunoassay (CEIA),
radioimmunoassay (RIA), immunofluorescence, chemiluminescence
immunoassay (CL), or electrochemiluminescence immunoassay (ECL). In
some embodiments, phosphorylation is detected and/or quantified
using an immunoassay that utilizes an amplified luminescent
proximity homogenous assay (AlphaLISA.RTM., PerkinElmer Inc.).
[0255] In some embodiments, phosphorylation is measured using a
sample that comprises one or more cells, e.g., one or more
TREM2-expressing cells (e.g., a primary cell or cell line that
endogenously expresses TREM2, such as human macrophages or
iPSC-derived microglia, or a primary cell or cell line that has
been engineered to express TREM2, e.g., as described in the
Examples section below). In some embodiments, the sample comprises
a fluid, e.g., blood, plasma, serum, urine, or cerebrospinal fluid.
In some embodiments, the sample comprises tissue (e.g., lung,
brain, kidney, spleen, nervous tissue, or skeletal muscle) or cells
from such tissue. In some embodiments, the sample comprises
endogenous fluid, tissue, or cells (e.g., from a human or non-human
subject).
[0256] Modulation of Phagocytosis
[0257] In some embodiments, an anti-TREM2 antibody enhances
phagocytosis of dead cell debris, tissue debris, amyloid beta
particles, or foreign material. In some embodiments, an anti-TREM2
antibody enhances phagocytosis without blocking binding of a native
TREM2 ligand. In some embodiments, an anti-TREM2 antibody enhances
phagocytosis that is induced by a TREM2 ligand (e.g., a lipid
ligand). In some embodiments, an anti-TREM2 antibody enhances
phagocytosis if the level of phagocytosis in a sample treated with
the anti-TREM2 antibody is increased by at least 10%, at least 20%,
at least 30%, at least 40%, at least 50%, at least 60%, at least
70%, at least 80%, at least 90% or more as compared to a control
value. In some embodiments, an anti-TREM2 antibody enhances
phagocytosis if the level of phagocytosis in a sample treated with
the anti-TREM2 antibody is increased by at least 2-fold, 3-fold,
4-fold, 5-fold, 6-fold, 7-fold, 8-fold, 9-fold, 10-fold, or more as
compared to a control value. In some embodiments, the control value
is the level of phagocytosis in an untreated sample, a sample that
has been treated with a TREM2 ligand but not an anti-TREM2
antibody, or a sample treated with an appropriate non-TREM2-binding
antibody.
[0258] In some embodiments, phagocytosis is measured using a
phagocytosis assay with a labeled substrate. Phagocytosis assays
are known in the art. In some embodiments, the phagocytosis assay
is performed on a sample comprising cells that endogenously express
TREM2, such as human macrophages or microglia. In some embodiments,
the phagocytosis assay is performed on a sample comprising cells
that have been engineered to express TREM2. In some embodiments,
phagocytosis is measured using a human macrophage phagocytosis
assay as described in the Examples section below.
[0259] Modulation of Cell Differentiation, Function, Migration, and
Survival
[0260] In some embodiments, an anti-TREM2 antibody enhances cell
migration, cell survival, cell function, or cell differentiation
(e.g., for myeloid cells, macrophages, and microglia, including
iPSC-derived microglia and disease-associated microglia).
Disease-associated microglia and methods of detecting
disease-associated microglia are described in Keren-Shaul et al.,
Cell, 2017, 169:1276-1290. In some embodiments, an anti-TREM2
antibody enhances cell migration of one or more cell types (e.g.,
myeloid cells, macrophages, or microglia). In some embodiments, an
anti-TREM2 antibody enhances cell survival of one or more cell
types (e.g., myeloid cells, macrophages, or microglia). In some
embodiments, an anti-TREM2 antibody enhances cell function of one
or more cell types (e.g., myeloid cells, macrophages, or
microglia). In some embodiments, an anti-TREM2 antibody enhances
cell differentiation of one or more cell types (e.g., myeloid
cells, macrophages, or microglia). In some embodiments, an
anti-TREM2 antibody enhances the migration, survival, function,
and/or differentiation of myeloid cells. In some embodiments, an
anti-TREM2 antibody enhances the migration, survival, function,
and/or differentiation of macrophages. In some embodiments, an
anti-TREM2 antibody enhances the migration, survival, function,
and/or differentiation of microglia. In some embodiments, an
anti-TREM2 antibody enhances microglia activation. In some
embodiments, an anti-TREM2 antibody enhances the migration,
survival, function, and/or differentiation of disease-associated
microglia. In some embodiments, an anti-TREM2 antibody enhances
cell migration, cell survival, cell function, or cell
differentiation without blocking binding of a native TREM2 ligand.
In some embodiments, an anti-TREM2 antibody enhances cell
migration, cell survival, cell function, or cell differentiation
that is induced by a TREM2 ligand (e.g., a lipid ligand).
[0261] In some embodiments, an anti-TREM2 antibody enhances cell
migration, cell survival, cell function, or cell differentiation if
the level of activity (e.g., migration, survival, function, or
differentiation) in a sample treated with the anti-TREM2 antibody
is increased by at least 10%, at least 20%, at least 30%, at least
40%, at least 50%, at least 60%, at least 70%, at least 80%, at
least 90% or more as compared to a control value. In some
embodiments, an anti-TREM2 antibody enhances cell migration, cell
survival, cell function, or cell differentiation if the level of
activity (e.g., migration, survival, function, or differentiation)
in a sample treated with the anti-TREM2 antibody is increased by at
least 2-fold, 3-fold, 4-fold, 5-fold, 6-fold, 7-fold, 8-fold,
9-fold, 10-fold, or more as compared to a control value. In some
embodiments, the control value is the level of activity (e.g.,
migration, survival, function, or differentiation) in an untreated
sample (e.g., a sample that has not been treated with an anti-TREM2
antibody), a sample that has been treated with a TREM2 ligand but
not an anti-TREM2 antibody, or a sample treated with an appropriate
non-TREM2-binding antibody.
[0262] In some embodiments, cell migration is measured using a
chemotaxis assay. Chemotaxis assays are known in the art. In some
embodiments, the cell migration assay (e.g., chemotaxis assay) is
performed on a sample comprising cells that endogenously express
TREM2, such as human macrophages. In some embodiments, the cell
migration assay (e.g., chemotaxis assay) is performed on a sample
comprising cells that have been engineered to express TREM2. In
some embodiments, cell migration is measured using a human
macrophage chemotaxis assay as described in the Examples section
below.
[0263] In some embodiments, cell survival is measured using a cell
viability assay. Cell viability assays are known in the art. In
some embodiments, the cell survival assay (e.g., cell viability
assay) is performed on a sample comprising cells that endogenously
express TREM2, such as human macrophages. In some embodiments, the
cell survival assay (e.g., cell viability assay) is performed on a
sample comprising cells that have been engineered to express TREM2.
In some embodiments, cell survival is measured using a human
macrophage viability assay as described in the Examples section
below.
[0264] In some embodiments, cell function is measured using a
functional assay that is appropriate for that cell. For example, in
some embodiments, macrophage cell function is evaluated using a
phagocytosis assay, e.g., as described in the Examples section
below.
[0265] In some embodiments, cell differentiation is measured by
evaluating the ability of cells that endogenously express TREM2 to
differentiate. For example, in some embodiments, cell
differentiation is measured by evaluating the ability of
macrophages to differentiate from monocytes, e.g., as described in
the Examples section below.
[0266] In some embodiments, activation of microglia is measured in
vivo. In some embodiments, microglia activation is measured using
TSPO-PET imaging. TSPO-PET imaging methods are known in the
art.
[0267] In some embodiments, an anti-TREM2 antibody enhances
microglia function without increasing neuroinflammation. Levels of
neuroinflammation can be determined by measuring levels of
cytokines (e.g., inflammatory cytokines), such as but not limited
to TNF-.alpha., IL-1.beta., IL-6, IL-1ra, TGF.beta., IL-15, or
IFN-.gamma.. In some embodiments, cytokine levels are measured
using immunoassays, for example, an enzyme immunoassay (EIA),
enzyme multiplied immunoassay (EMIA), enzyme-linked immunosorbent
assay (ELISA), microparticle enzyme immunoassay (MEIA),
immunohistochemistry (IHC), immunocytochemistry, capillary
electrophoresis immunoassay (CEIA), radioimmunoassay (RIA),
immunofluorescence, chemiluminescence immunoassay (CL), or
electrochemiluminescence immunoassay (ECL).
IV. FC Polypeptide Mutations of Proteins Having Anti-TREM2 Antigen
Binding Portion
[0268] In some aspects, an anti-TREM2 antibody comprises two Fc
polypeptides, one or both of which may each comprise independently
selected modifications (e.g., mutations) or may be a wild-type Fc
polypeptide, e.g., a human IgG1 Fc polypeptide. Non-limiting
examples of mutations that can be introduced into one or both Fc
polypeptides include, e.g., mutations to permit binding of an Fc
polypeptide (or antibody comprising the same) to a BBB-receptor,
such as transferrin receptor (TfR) protein (e.g., a human or
cynomolgus TfR, such as may be expressed on a brain endothelial
cell), mutations to increase serum stability, to modulate effector
function, to influence glycosylation, to reduce immunogenicity in
humans, and/or to provide for knob and hole heterodimerization of
the Fc polypeptides.
Transferrin Receptor-Binding Mutations
[0269] In some embodiments, an anti-TREM2 antibody includes an Fc
polypeptide that comprises modifications (e.g., amino acid
substitutions) that permit binding of the Fc polypeptide to a TfR
protein. Briefly, binding to a TfR protein (e.g., to the apical
domain thereof) that is expressed on, for example, a brain
endothelial cell, can, in some embodiments, permit a modified Fc
polypeptide of this disclosure or an antibody comprising the same
to cross the blood-brain barrier via receptor-mediated
transcytosis. In certain embodiments, receptor-mediated
transcytosis can enhance or improve the ability of the protein
comprising the Fc polypeptide to be present in the brain (i.e., on
the luminal side of the blood-brain barrier), which can allow for
improved binding to TREM2 in the CNS, and other functions, e.g.,
clearance, neutralization, or immunodepletion of the target, or the
like.
[0270] Exemplary TfR-binding amino acid modifications to an Fc
(e.g., CH2 and/or CH3 portion, fragment, or domain), and Fc
polypeptides and portions thereof that comprise the amino acid
modifications, are described in PCT patent publication no. WO
2018/152326A1. These amino acid modifications, TfR-binding Fc
polypeptide sequences and TfR-binding Fc polypeptides, and
techniques for generating and testing the same are incorporated
herein by reference. One or two Fc polypeptides of an Fc dimer of
the present disclosure can be engineered to comprise modifications
to permit binding to TfR. In certain embodiments, one Fc
polypeptide of an Fc dimer comprises modifications to permit
binding to TfR, and the other Fc polypeptide does not.
[0271] In some embodiments, a modified Fc polypeptide comprises a
YxTEWSS (SEQ ID NO:58) motif. In some embodiments, a modified Fc
polypeptide comprises a TxxExxxxF (SEQ ID NO:59) motif. In some
embodiments, a modified Fc polypeptide comprises a YxTEWSS (SEQ ID
NO:58) and a TxxExxxxF (SEQ ID NO:59) motif.
[0272] In some embodiments, a modified Fc polypeptide comprises a
wild-type amino acid residue at positions 380, 389, 390, and 415,
according to EU numbering, wherein the wild-type amino acid residue
is found at a corresponding position in SEQ ID NO:38.
[0273] In some embodiments, an anti-TREM2 antibody includes an Fc
polypeptide having the following amino acids: Trp, Leu, or Glu at
position 380; Tyr or Phe at position 384; Thr at position 386; Glu
at position 387; Trp at position 388; Ser, Ala, or Val at position
389; Ser or Asn at position 390; Thr or Ser at position 413; Glu or
Ser at position 415; Glu at position 416; and Phe at position 421,
according to EU numbering.
[0274] In some embodiments, the anti-TREM2 antibody includes an Fc
polypeptide having the following amino acids: Trp at position 380;
Tyr at position 384; Thr at position 386; Glu at position 387; Trp
at position 388; Ser at position 389; Ser at position 390; Thr at
position 413; Glu at position 415; Glu at position 416; and Phe at
position 421, according to EU numbering.
[0275] In some embodiments, an anti-TREM2 antibody includes an Fc
polypeptide having the following amino acids: Glu at position 380;
Phe at position 384; Thr at position 386; Glu at position 387; Trp
at position 388; Ser at position 389; Asn at position 390; Ser at
position 413; Glu at position 415; Glu at position 416; and Phe at
position 421, according to EU numbering.
[0276] In some embodiments, an anti-TREM2 antibody includes an Fc
polypeptide having the following amino acids: Glu at position 380;
Tyr at position 384; Thr at position 386; Glu at position 387; Trp
at position 388; Val at position 389; Asn at position 390; Thr at
position 413; Glu at position 415; Glu at position 416; and Phe at
position 421, according to EU numbering.
[0277] In some embodiments, an anti-TREM2 antibody includes an Fc
polypeptide having the following amino acids: Glu at position 380;
Tyr at position 384; Thr at position 386; Glu at position 387; Trp
at position 388; Ser at position 389; Asn at position 390; Ser at
position 413; Glu at position 415; Glu at position 416; and Phe at
position 421, according to EU numbering.
[0278] In some embodiments, a modified Fc polypeptide comprises a
sequence having at least 90% identity to an amino acid sequence set
forth in any one of SEQ ID NOS:40, 43, 46, and 49. In some
embodiments, a modified Fc polypeptide comprises or consists of the
amino acid sequence set forth in any one of SEQ ID NOS:41, 44, 47,
and 50.
[0279] Additional examples of modified Fc polypeptides are
described in Table 8.
Mutations to Promote Heterodimerization of Fc Polypeptides
[0280] In some embodiments, the Fc polypeptides present in an
anti-TREM2 antibody as disclosed herein include knob and hole
mutations to promote heterodimer formation and hinder homodimer
formation. Generally, the modifications introduce a protuberance
("knob") at the interface of a first polypeptide and a
corresponding cavity ("hole") in the interface of a second
polypeptide, such that the protuberance can be positioned in the
cavity so as to promote heterodimer formation and thus hinder
homodimer formation. Protuberances are constructed by replacing
small amino acid side chains from the interface of the first
polypeptide with larger side chains (e.g., tyrosine or tryptophan).
Compensatory cavities of identical or similar size to the
protuberances are created in the interface of the second
polypeptide by replacing large amino acid side chains with smaller
ones (e.g., alanine or threonine). In some embodiments, such
additional mutations are at a position in the Fc polypeptide that
does not have a negative (e.g., inhibitory) effect on binding of a
Fc polypeptide to a BBB receptor, e.g., TfR.
[0281] In one illustrative embodiment of a knob and hole approach
for dimerization, position 366 (numbered according to the EU
numbering scheme) of one of the Fc polypeptides present in the
proteins described herein comprises a tryptophan in place of a
native threonine. The other Fc polypeptide in the dimer has a
valine at position 407 (numbered according to the EU numbering
scheme) in place of the native tyrosine. The other Fc polypeptide
may further comprise a substitution in which the native threonine
at position 366 (numbered according to the EU numbering scheme) is
substituted with a serine and a native leucine at position 368
(numbered according to the EU numbering scheme) is substituted with
an alanine. Thus, one of the Fc polypeptides of an anti-TREM2
protein of the disclosure has the T366W knob mutation and the other
Fc polypeptide has the Y407V mutation, which is typically
accompanied by the T366S and L368A hole mutations.
[0282] In some embodiments, one or both Fc polypeptides may also be
engineered to contain other modifications for heterodimerization,
e.g., electrostatic engineering of contact residues within a
CH3-CH3 interface that are naturally charged or hydrophobic patch
modifications.
[0283] In some embodiments, modifications to enhance serum
half-life may be introduced. For example, in some embodiments, one
or both Fc polypeptides present in an anti-TREM2 protein of the
disclosure may comprise a tyrosine at position 252, a threonine at
position 254, and a glutamic acid at position 256, as numbered
according to the EU numbering scheme. Thus, one or both Fc
polypeptides may have M252Y, S254T, and T256E substitutions.
Alternatively, one or both Fc polypeptides may have M428L and/or
N434S substitutions, according to EU numbering. Alternatively, one
or both Fc polypeptides may have an N434S or N434A
substitution.
Fc Effector Functions
[0284] In some embodiments, one or both Fc polypeptides in an
anti-TREM2 protein disclosed herein may comprise modifications that
reduce effector function, i.e., having a reduced ability to induce
certain biological functions upon binding to an Fc receptor
expressed on an effector cell that mediates the effector function.
Examples of antibody effector functions include, but are not
limited to, C1q binding and complement dependent cytotoxicity
(CDC), Fc receptor binding, antibody-dependent cell-mediated
cytotoxicity (ADCC), antibody-dependent cell-mediated phagocytosis
(ADCP), down-regulation of cell surface receptors (e.g., B cell
receptor), and B-cell activation. Effector functions may vary with
the antibody class. For example, native human IgG1 and IgG3
antibodies can elicit ADCC and CDC activities upon binding to an
appropriate Fc receptor present on an immune system cell; and
native human IgG1, IgG2, IgG3, and IgG4 can elicit ADCP functions
upon binding to the appropriate Fc receptor present on an immune
cell.
[0285] In some embodiments, one or both Fc polypeptides may include
modifications that modulate effector function.
[0286] In some embodiments, one or both Fc polypeptides may
comprise modifications that reduce or eliminate effector function.
Illustrative Fc polypeptide mutations that reduce effector function
include, but are not limited to, substitutions in a CH2 domain,
e.g., at positions 234 and 235, according to the EU numbering
scheme. For example, in some embodiments, one or both Fc
polypeptides can comprise alanine residues at positions 234 and
235. Thus, one or both Fc polypeptides may have L234A and L235A
(LALA) substitutions.
[0287] Additional Fc polypeptide mutations that modulate an
effector function include, but are not limited to, the following:
position 329 may have a mutation in which proline is substituted
with a glycine, arginine, serine, or an amino acid residue large
enough to destroy the Fc/Fc.gamma. receptor interface that is
formed between proline 329 of the Fc and tryptophan residues Trp 87
and Trp 110 of FcTRIII. Additional illustrative substitutions
include S228P, E233P, L235E, N297A, N297D, N297G, and P331S,
according to the EU numbering scheme. Multiple substitutions may
also be present, e.g., L234A and L235A of a human IgG1 Fc region;
L234A, L235A, and P329G of a human IgG1 Fc region; L234A, L235A,
and P329S of a human IgG1 Fc region; S228P and L235E of a human
IgG4 Fc region; L234A and G237A of a human IgG1 Fc region; L234A,
L235A, and G237A of a human IgG1 Fc region; V234A and G237A of a
human IgG2 Fc region; L235A, G237A, and E318A of a human IgG4 Fc
region; and S228P and L236E of a human IgG4 Fc region, according to
the EU numbering scheme. In some embodiments, one or both Fc
polypeptides may have one or more amino acid substitutions that
modulate ADCC, e.g., substitutions at positions 298, 333, and/or
334, according to the EU numbering scheme.
FcRn Binding Sites and Mutations to Increase Serum Half-Life
[0288] In certain aspects, Fc polypeptides (e.g., modified Fc
polypeptides) present in an anti-TREM2 protein of the disclosure,
can comprise an FcRn binding site. In some embodiments, the FcRn
binding site is within the Fc polypeptide or a fragment
thereof.
[0289] In some embodiments, the FcRn binding site comprises a
native FcRn binding site. In some embodiments, the FcRn binding
site does not comprise amino acid changes relative to the amino
acid sequence of a native FcRn binding site. In some embodiments,
the native FcRn binding site is an IgG binding site, e.g., a human
IgG binding site. In some embodiments, the FcRn binding site
comprises a modification that alters FcRn binding.
[0290] In some embodiments, an FcRn binding site has one or more
amino acid residues that are mutated, e.g., substituted, wherein
the mutation(s) increase serum half-life or do not substantially
reduce serum half-life (i.e., reduce serum half-life by no more
than 25% compared to a counterpart Fc polypeptide having the
wild-type residues at the mutated positions when assayed under the
same conditions). In some embodiments, an FcRn binding site has one
or more amino acid residues that are substituted at positions
251-256, 428, and 433-436, according to the EU numbering
scheme.
[0291] In some embodiments, one or more residues at or near an FcRn
binding site are mutated, relative to a native human IgG sequence,
to extend serum half-life of the polypeptide. In some embodiments,
mutations are introduced into one, two, or three of positions 252,
254, and 256. In some embodiments, the mutations are M252Y, S254T,
and T256E. In some embodiments, an Fc polypeptide further comprises
the mutations M252Y, S254T, and T256E. In particular embodiments,
one or both Fc polypeptides present in an anti-TREM2 protein of the
disclosure may comprise a tyrosine at position 252, a threonine at
position 254, and a glutamic acid at position 256, as numbered
according to the EU numbering scheme. Thus, one or both Fc
polypeptides may have M252Y, S254T, and T256E substitutions.
[0292] In some embodiments, the mutations are M428L and/or N434S.
In some embodiments, an Fc polypeptide further comprises the
mutation N434S with or without M428L. In some embodiments, an Fc
polypeptide comprises a mutation at one, two, or all three of
positions T307, E380, and N434, according to the EU numbering
scheme. In some embodiments, the mutations are T307Q and N434A. In
some embodiments, an Fc polypeptide comprises mutations T307A,
E380A, and N434A. In some embodiments, an Fc polypeptide comprises
mutations at positions T250 and M428, according to the EU numbering
scheme. In some embodiments, the Fc polypeptide comprises mutations
T250Q and/or M428L. In some embodiments, an Fc polypeptide
comprises mutations at positions M428 and N434, according to the EU
numbering scheme. In some embodiments, the Fc polypeptide comprises
mutations M428L and N434S. In some embodiments, an antibody of the
present disclosure can comprise two Fc polypeptides, wherein each
of the two Fc polypeptides comprises M428L and/or N434S
substitutions. In some embodiments, the Fc polypeptide comprises an
N434S or N434A mutation. In some embodiments, an antibody of the
present disclosure can comprise two Fc polypeptides, wherein each
of the two Fc polypeptides comprises an N434S or N434A
substitution.
V. Preparation of Antibodies
[0293] In some embodiments, antibodies are prepared by immunizing
an animal or animals (e.g., mice, rabbits, or rats) with an antigen
or a mixture of antigens for the induction of an antibody response.
In some embodiments, the antigen or mixture of antigens is
administered in conjugation with an adjuvant (e.g., Freund's
adjuvant). After an initial immunization, one or more subsequent
booster injections of the antigen or antigens may be administered
to improve antibody production. Following immunization,
antigen-specific B cells are harvested, e.g., from the spleen
and/or lymphoid tissue. For generating monoclonal antibodies, the B
cells are fused with myeloma cells, which are subsequently screened
for antigen specificity. Methods of preparing antibodies are also
described in the Examples section below.
[0294] The genes encoding the heavy and light chains of an antibody
of interest can be cloned from a cell, e.g., the genes encoding a
monoclonal antibody can be cloned from a hybridoma and used to
produce a recombinant monoclonal antibody. Gene libraries encoding
heavy and light chains of monoclonal antibodies can also be made
from hybridoma or plasma cells. Alternatively, phage or yeast
display technology can be used to identify antibodies and Fab
fragments that specifically bind to selected antigens. Antibodies
can also be made bispecific, i.e., able to recognize two different
antigens. Antibodies can also be heteroconjugates, e.g., two
covalently joined antibodies, or immunotoxins.
[0295] Antibodies can be produced using any number of expression
systems, including prokaryotic and eukaryotic expression systems.
In some embodiments, the expression system is a mammalian cell
expression, such as a hybridoma, or a CHO cell expression system.
Many such systems are widely available from commercial suppliers.
In embodiments in which an antibody comprises both a V.sub.H and
V.sub.L region, the V.sub.H and V.sub.L regions may be expressed
using a single vector, e.g., in a di-cistronic expression unit, or
under the control of different promoters. In other embodiments, the
V.sub.H and V.sub.L region may be expressed using separate vectors.
A V.sub.H or V.sub.L region as described herein may optionally
comprise a methionine at the N-terminus.
[0296] In some embodiments, the antibody is a chimeric antibody.
Methods for making chimeric antibodies are known in the art. For
example, chimeric antibodies can be made in which the antigen
binding region (heavy chain variable region and light chain
variable region) from one species, such as a mouse, is fused to the
effector region (constant domain) of another species, such as a
human. As another example, "class switched" chimeric antibodies can
be made in which the effector region of an antibody is substituted
with an effector region of a different immunoglobulin class or
subclass.
[0297] In some embodiments, the antibody is a humanized antibody.
Generally, a non-human antibody is humanized in order to reduce its
immunogenicity. Humanized antibodies typically comprise one or more
variable regions (e.g., CDRs) or portions thereof that are
non-human (e.g., derived from a mouse variable region sequence),
and possibly some framework regions or portions thereof that are
non-human, and further comprise one or more constant regions that
are derived from human antibody sequences. Methods for humanizing
non-human antibodies are known in the art. Transgenic mice, or
other organisms such as other mammals, can be used to express
humanized or human antibodies. Other methods of humanizing
antibodies include, for example, variable domain resurfacing, CDR
grafting, grafting specificity-determining residues (SDR), guided
selection, and framework shuffling.
[0298] As an alternative to humanization, fully human antibodies
can be generated. As a non-limiting example, transgenic animals
(e.g., mice) can be produced that are capable, upon immunization,
of producing a full repertoire of human antibodies in the absence
of endogenous immunoglobulin production. For example, it has been
described that the homozygous deletion of the antibody heavy-chain
joining region (JH) gene in chimeric and germ-line mutant mice
results in complete inhibition of endogenous antibody production.
Transfer of the human germ-line immunoglobulin gene array in such
germ-line mutant mice will result in the production of human
antibodies upon antigen challenge. As another example, human
antibodies can be produced by hybridoma-based methods, such as by
using primary human B cells for generating cell lines producing
human monoclonal antibodies.
[0299] Human antibodies can also be produced using phage display or
yeast display technology. In phage display, repertoires of variable
heavy chain and variable light chain genes are amplified and
expressed in phage display vectors. In some embodiments, the
antibody library is a natural repertoire amplified from a human
source. In some embodiments, the antibody library is a synthetic
library made by cloning heavy chain and light chain sequences and
recombining to generate a large pool of antibodies with different
antigenic specificity. Phage typically display antibody fragments
(e.g., Fab fragments or scFv fragments), which are then screened
for binding to an antigen of interest.
[0300] In some embodiments, antibody fragments (such as a Fab, a
Fab', a F(ab').sub.2, a scFv, a V.sub.H, or a V.sub.HH) are
generated. Various techniques have been developed for the
production of antibody fragments. Traditionally, these fragments
were derived via proteolytic digestion of intact antibodies.
However, these fragments can now be produced directly using
recombinant host cells. For example, antibody fragments can be
isolated from antibody phage libraries. Alternatively, Fab'-SH
fragments can be directly recovered from E. coli cells and
chemically coupled to form F(ab').sub.2 fragments. According to
another approach, F(ab').sub.2 fragments can be isolated directly
from recombinant host cell culture. Other techniques for the
production of antibody fragments will be apparent to those skilled
in the art.
[0301] In some embodiments, an antibody or an antibody fragment is
conjugated to another molecule, e.g., polyethylene glycol
(PEGylation) or serum albumin, to provide an extended half-life in
vivo.
VI. Nucleic Acids, Vectors, and Host Cells
[0302] In some embodiments, the anti-TREM2 antibodies as disclosed
herein are prepared using recombinant methods. Accordingly, in some
aspects, the disclosure provides isolated nucleic acids comprising
a nucleic acid sequence encoding any of the anti-TREM2 antibodies
as described herein (e.g., any one or more of the CDRs, heavy chain
variable regions, and light chain variable regions described
herein); vectors comprising such nucleic acids; and host cells into
which the nucleic acids are introduced that are used to replicate
the antibody-encoding nucleic acids and/or to express the
antibodies.
[0303] In some embodiments, a polynucleotide (e.g., an isolated
polynucleotide) comprises a nucleotide sequence encoding an
antibody as described herein (e.g., as described in the Section
above entitled "Anti-TREM2 Antibody Sequences"). In some
embodiments, the polynucleotide comprises a nucleotide sequence
encoding one or more amino acid sequences (e.g., CDR, heavy chain,
or light chain sequences) disclosed in Table 8 below. In some
embodiments, the polynucleotide comprises a nucleotide sequence
encoding an amino acid sequence having at least 85% sequence
identity (e.g., at least 85%, at least 90%, at least 91%, at least
92%, at least 93%, at least 94%, at least 95%, at least 96%, at
least 97%, at least 98%, or at least 99% sequence identity) to a
sequence (e.g., a CDR, heavy chain, or light chain sequence)
disclosed in Table 8 below. In some embodiments, a polynucleotide
as described herein is operably linked to a heterologous nucleic
acid, e.g., a heterologous promoter.
[0304] Suitable vectors containing polynucleotides encoding
antibodies of the present disclosure, or fragments thereof, include
cloning vectors and expression vectors. While the cloning vector
selected may vary according to the host cell intended to be used,
useful cloning vectors generally have the ability to
self-replicate, may possess a single target for a particular
restriction endonuclease, and/or may carry genes for a marker that
can be used in selecting clones containing the vector. Examples
include plasmids and bacterial viruses, e.g., pUC18, pUC19,
Bluescript (e.g., pBS SK+) and its derivatives, mpl8, mpl9, pBR322,
pMB9, ColEl, pCR1, RP4, phage DNAs, and shuttle vectors such as
pSA3 and pAT28. These and many other cloning vectors are available
from commercial vendors such as BioRad, Strategene, and
Invitrogen.
[0305] Expression vectors generally are replicable polynucleotide
constructs that contain a nucleic acid of the present disclosure.
The expression vector may replicate in the host cells either as
episomes or as an integral part of the chromosomal DNA. Suitable
expression vectors include but are not limited to plasmids, viral
vectors, including adenoviruses, adeno-associated viruses,
retroviruses, and any other vector.
[0306] Suitable host cells for cloning or expressing a
polynucleotide or vector as described herein include prokaryotic or
eukaryotic cells. In some embodiments, the host cell is
prokaryotic. In some embodiments, the host cell is eukaryotic,
e.g., Chinese Hamster Ovary (CHO) cells or lymphoid cells. In some
embodiments, the host cell is a human cell, e.g., a Human Embryonic
Kidney (HEK) cell.
[0307] In another aspect, methods of making an anti-TREM2 antibody
as described herein are provided. In some embodiments, the method
includes culturing a host cell as described herein (e.g., a host
cell expressing a polynucleotide or vector as described herein)
under conditions suitable for expression of the antibody. In some
embodiments, the antibody is subsequently recovered from the host
cell (or host cell culture medium).
VII. Therapeutic Methods Using Anti-TREM2 Antibodies
[0308] In another aspect, therapeutic methods using an anti-TREM2
antibody as disclosed herein (e.g., an anti-TREM2 antibody as
described in Section III above) are provided. In some embodiments,
methods of treating a neurodegenerative disease are provided. In
some embodiments, methods of modulating one or more TREM2
activities (e.g., in a subject having a neurodegenerative disease)
are provided.
[0309] In some embodiments, methods of treating a neurodegenerative
disease are provided. In some embodiments, the neurodegenerative
disease is selected from the group consisting of Alzheimer's
disease, primary age-related tauopathy, progressive supranuclear
palsy (PSP), frontotemporal dementia, frontotemporal dementia with
parkinsonism linked to chromosome 17, argyrophilic grain dementia,
amyotrophic lateral sclerosis, amyotrophic lateral
sclerosis/parkinsonism-dementia complex of Guam (ALS-PDC),
corticobasal degeneration, chronic traumatic encephalopathy,
Creutzfeldt-Jakob disease, dementia pugilistica, diffuse
neurofibrillary tangles with calcification, Down's syndrome,
familial British dementia, familial Danish dementia,
Gerstmann-Straussler-Scheinker disease, globular glial tauopathy,
Guadeloupean parkinsonism with dementia, Guadelopean PSP,
Hallevorden-Spatz disease, hereditary diffuse leukoencephalopathy
with spheroids (HDLS), Huntington's disease, inclusion-body
myositis, multiple system atrophy, myotonic dystrophy, Nasu-Hakola
disease, neurofibrillary tangle-predominant dementia, Niemann-Pick
disease type C, pallido-ponto-nigral degeneration, Parkinson's
disease, Pick's disease, postencephalitic parkinsonism, prion
protein cerebral amyloid angiopathy, progressive subcortical
gliosis, subacute sclerosing panencephalitis, and tangle only
dementia. In some embodiments, the neurodegenerative disease is
Alzheimer's disease. In some embodiments, the neurodegenerative
disease is Nasu-Hakola disease. In some embodiments, the
neurodegenerative disease is frontotemporal dementia. In some
embodiments, the neurodegenerative disease is Parkinson's disease.
In some embodiments, the method comprises administering to the
subject an isolated antibody or an antigen-binding fragment thereof
that specifically binds to a human TREM2 protein, e.g., an
anti-TREM2 antibody as described herein, or a pharmaceutical
composition comprising an anti-TREM2 antibody as described
herein.
[0310] In some embodiments, an anti-TREM2 antibody (or
antigen-binding portion or pharmaceutical composition thereof) as
described herein is used in treating a neurodegenerative disease
that is characterized by a mutation in TREM2. In some embodiments,
the neurodegenerative disease that is characterized by a mutation
in TREM2 is Alzheimer's disease, e.g., Alzheimer's disease that is
characterized by a R47H mutation in TREM2.
[0311] In some embodiments, methods of modulating one or more TREM2
activities in a subject (e.g., a subject having a neurodegenerative
disease) are provided. In some embodiments, the method comprises
modulating levels of sTREM2; modulating recruitment or
phosphorylation of a kinase that interacts with a TREM2/DAP12
signaling complex (e.g., Syk kinase); modulating phagocytosis
(e.g., phagocytosis of cell debris, amyloid beta particles, etc.);
modulating cell migration (e.g., migration of myeloid cells,
macrophages, microglia, and disease associated microglia); and/or
modulating cell differentiation (e.g., for myeloid cells,
macrophages, microglia, and disease associated microglia). In some
embodiments, methods of enhancing one or more TREM2 activities in a
subject having a neurodegenerative disease are provided. In some
embodiments, methods of decreasing levels of sTREM2 in a subject
having a neurodegenerative disease are provided. In some
embodiments, the method of modulating one or more TREM2 activities
in a subject comprises administering to the subject an isolated
antibody or an antigen-binding portion thereof that specifically
binds to a human TREM2 protein, e.g., an anti-TREM2 antibody as
describe herein, or a pharmaceutical composition comprising an
anti-TREM2 antibody as described herein.
[0312] In some embodiments, the subject to be treated is a human,
e.g., a human adult or a human child.
[0313] In some embodiments, methods of reducing plaque accumulation
in a subject having a neurodegenerative disease are provided. In
some embodiments, the method comprises administering to the subject
an antibody or pharmaceutical composition as described herein. In
some embodiments, the subject has Alzheimer's disease. In some
embodiments, the subject is an animal model of a neurodegenerative
disease (e.g., a 5XFAD or APP/PS1 mouse model). In some
embodiments, plaque accumulation is measured by amyloid plaque
imaging and/or Tau imaging, e.g., using positron emission
tomography (PET) scanning. In some embodiments, administration of
an anti-TREM2 antibody reduces plaque accumulation by at least 20%,
at least 30%, at least 40%, at least 50%, at least 60%, at least
70%, at least 80%, or at least 90% as compared to a baseline value
(e.g., the level of plaque accumulation in the subject prior to
administration of the anti-TREM2 antibody).
[0314] In some embodiments, an anti-TREM2 antibody is administered
to a subject at a therapeutically effective amount or dose. The
dosages, however, may be varied according to several factors,
including the chosen route of administration, the formulation of
the composition, patient response, the severity of the condition,
the subject's weight, and the judgment of the prescribing
physician. The dosage can be increased or decreased over time, as
required by an individual patient. In certain instances, a patient
initially is given a low dose, which is then increased to an
efficacious dosage tolerable to the patient. Determination of an
effective amount is well within the capability of those skilled in
the art.
[0315] The route of administration of an anti-TREM2 antibody as
described herein can be oral, intraperitoneal, transdermal,
subcutaneous, intravenous, intramuscular, intrathecal,
inhalational, topical, intralesional, rectal, intrabronchial,
nasal, transmucosal, intestinal, ocular or otic delivery, or any
other methods known in the art. In some embodiments, the antibody
is administered orally, intravenously, or intraperitoneally.
[0316] In some embodiments, the anti-TREM2 antibody (and optionally
another therapeutic agent) is administered to the subject over an
extended period of time, e.g., for at least 30, 40, 50, 60, 70, 80,
90, 100, 150, 200, 250, 300, 350 days or longer.
VIII. Pharmaceutical Compositions and Kits
[0317] In another aspect, pharmaceutical compositions and kits
comprising an antibody that specifically binds to a human TREM2
protein are provided. In some embodiments, the pharmaceutical
compositions and kits are for use in treating a neurodegenerative
disease. In some embodiments, the pharmaceutical compositions and
kits are for use in modulating (e.g., enhancing or inhibiting) one
or more TREM2 activities, e.g., Syk phosphorylation. In some
embodiments, the pharmaceutical compositions and kits are for use
in modulating (e.g., decreasing) sTREM2 levels.
Pharmaceutical Compositions
[0318] In some embodiments, pharmaceutical compositions comprising
an anti-TREM2 antibody or an antigen-binding fragment thereof are
provided. In some embodiments, the anti-TREM2 antibody is an
antibody as described in Section III above or an antigen-binding
fragment thereof.
[0319] In some embodiments, a pharmaceutical composition comprises
an anti-TREM2 antibody as described herein and further comprises
one or more pharmaceutically acceptable carriers and/or excipients.
A pharmaceutically acceptable carrier includes any solvents,
dispersion media, or coatings that are physiologically compatible
and that does not interfere with or otherwise inhibit the activity
of the active agent. Various pharmaceutically acceptable excipients
are well-known in the art.
[0320] In some embodiments, the carrier is suitable for
intravenous, intramuscular, oral, intraperitoneal, intrathecal,
transdermal, topical, or subcutaneous administration.
Pharmaceutically acceptable carriers can contain one or more
physiologically acceptable compound(s) that act, for example, to
stabilize the composition or to increase or decrease the absorption
of the active agent(s). Physiologically acceptable compounds can
include, for example, carbohydrates, such as glucose, sucrose, or
dextrans, antioxidants, such as ascorbic acid or glutathione,
chelating agents, low molecular weight proteins, compositions that
reduce the clearance or hydrolysis of the active agents, or
excipients or other stabilizers and/or buffers. Other
pharmaceutically acceptable carriers and their formulations are
well-known in the art.
[0321] The pharmaceutical compositions described herein can be
manufactured in a manner that is known to those of skill in the
art, e.g., by means of conventional mixing, dissolving,
granulating, dragee-making, emulsifying, encapsulating, entrapping
or lyophilizing processes. The following methods and excipients are
merely exemplary and are in no way limiting.
[0322] For oral administration, an anti-TREM2 antibody can be
formulated by combining it with pharmaceutically acceptable
carriers that are well known in the art. Such carriers enable the
compounds to be formulated as tablets, pills, dragees, capsules,
emulsions, lipophilic and hydrophilic suspensions, liquids, gels,
syrups, slurries, suspensions and the like, for oral ingestion by a
patient to be treated. Pharmaceutical preparations for oral use can
be obtained by mixing the compounds with a solid excipient,
optionally grinding a resulting mixture, and processing the mixture
of granules, after adding suitable auxiliaries, if desired, to
obtain tablets or dragee cores. Suitable excipients include, for
example, fillers such as sugars, including lactose, sucrose,
mannitol, or sorbitol; cellulose preparations such as, for example,
maize starch, wheat starch, rice starch, potato starch, gelatin,
gum tragacanth, methyl cellulose, hydroxypropylmethyl-cellulose,
sodium carboxymethylcellulose, and/or polyvinylpyrrolidone (PVP).
If desired, disintegrating agents can be added, such as a
cross-linked polyvinyl pyrrolidone, agar, or alginic acid or a salt
thereof such as sodium alginate.
[0323] An anti-TREM2 antibody can be formulated for parenteral
administration by injection, e.g., by bolus injection or continuous
infusion. For injection, the compound or compounds can be
formulated into preparations by dissolving, suspending or
emulsifying them in an aqueous or nonaqueous solvent, such as
vegetable or other similar oils, synthetic aliphatic acid
glycerides, esters of higher aliphatic acids or propylene glycol;
and if desired, with conventional additives such as solubilizers,
isotonic agents, suspending agents, emulsifying agents, stabilizers
and preservatives. In some embodiments, compounds can be formulated
in aqueous solutions, e.g., in physiologically compatible buffers
such as Hanks's solution, Ringer's solution, or physiological
saline buffer. Formulations for injection can be presented in unit
dosage form, e.g., in ampules or in multi-dose containers, with an
added preservative. The compositions can take such forms as
suspensions, solutions or emulsions in oily or aqueous vehicles,
and can contain formulatory agents such as suspending, stabilizing
and/or dispersing agents.
[0324] Typically, a pharmaceutical composition for use in in vivo
administration is sterile. Sterilization can be accomplished
according to methods known in the art, e.g., heat sterilization,
steam sterilization, sterile filtration, or irradiation.
[0325] Dosages and desired drug concentration of pharmaceutical
compositions of the disclosure may vary depending on the particular
use envisioned. The determination of the appropriate dosage or
route of administration is well within the skill of one in the art.
Suitable dosages are also described in Section VII above.
Kits
[0326] In some embodiments, kits comprising an anti-TREM2 antibody
are provided. In some embodiments, the anti-TREM2 antibody is an
antibody as described in Section III above or an antigen-binding
fragment thereof.
[0327] In some embodiments, the kit further comprises one or more
additional therapeutic agents. For example, in some embodiments,
the kit comprises an anti-TREM2 antibody as described herein and
further comprises one or more additional therapeutic agents for use
in the treatment of a neurodegenerative disease, e.g., Alzheimer's
disease. In some embodiments, the therapeutic agent is an agent for
use in treating a cognitive or behavioral symptom of a
neurodegenerative disease (e.g., an antidepressant, a dopamine
agonist, or an anti-psychotic). In some embodiments, the
therapeutic agent is a neuroprotective agent (e.g.,
carbidopa/levodopa, an anticholinergic agent, a dopaminergic agent,
a monoamine oxidase B (MAO-B) inhibitor, a catechol-O-methyl
transferase (COMT) inhibitor, a glutamatergic agent, a histone
deacetylase (HDAC) inhibitor, a cannabinoid, a caspase inhibitor,
melatonin, an anti-inflammatory agent, a hormone (e.g., estrogen or
progesterone), or a vitamin).
[0328] In some embodiments, the kit comprises an anti-TREM2
antibody as described herein and further comprises one or more
reagents for measuring sTREM2 levels. In some embodiments, the kit
comprises an anti-TREM2 antibody as described herein and further
comprises one or more reagents for measuring TREM2 activity (e.g.,
for measuring Syk phosphorylation).
[0329] In some embodiments, the kit further comprises instructional
materials containing directions (i.e., protocols) for the practice
of the methods described herein (e.g., instructions for using the
kit for a therapeutic method as described in Section VI above).
While the instructional materials typically comprise written or
printed materials, they are not limited to such. Any medium capable
of storing such instructions and communicating them to an end user
is contemplated by this disclosure. Such media include, but are not
limited to, electronic storage media (e.g., magnetic discs, tapes,
cartridges, chips), optical media (e.g., CD-ROM), and the like.
Such media may include addresses to internet sites that provide
such instructional materials.
IX. Examples
[0330] The present disclosure will be described in greater detail
by way of specific examples. The following examples are offered for
illustrative purposes only, and are not intended to limit the
disclosure in any manner.
Example 1. Generation and Initial Characterization of Anti-TREM2
Antibodies
[0331] Recombinant Expression and Purification of Mouse Fc Fused
Human TREM2 ECD
[0332] The ecto domain (residues 19-172) of human TREM2 (UniProtKB
ID--Q9NZC2) was subcloned into pRK vector with the secretion signal
from mouse IgG kappa chain V-III, amino acids 1-20 (UniProtKB
ID--P01661) at the N-terminal region, and a mouse Fc tag at the
C-terminal region with a GGGGS (SEQ ID NO:34) between TREM2 ECD and
Fc.
[0333] Purified plasmid was transfected into Expi293F.TM. cells
(Thermo Fisher) using the Expi293F.TM. Expression System Kit
according to the manufacturer's instructions. To inhibit maturation
of N-linked glycans and reduce glycosylation heterogeneity,
kifunensine (Sigma), an inhibitor of high mannosidase I was added
to the culture at 1 .mu.g/mL concentration immediately after
transfection. Transfected cells were incubated in an orbital shaker
(Infors HT Multitron) at 125 rpm and 37.degree. C. in a humidified
atmosphere of 6% CO.sub.2. ExpiFectamine.TM. 293 Transfection
Enhancer 1 and 2 were added to the cells 16 hours post transfection
and the media supernatant was harvested 96 hours post transfection.
The clarified supernatant was supplemented with EDTA-free protease
inhibitor (Roche) and was stored at -80.degree. C.
[0334] For rhTREM2-Fc isolation, clarified media supernatant was
loaded on HiTrap MabSelect SuRe Protein A affinity column (GE
Healthcare Life Sciences) and washed with 200 mM arginine and 137
mM succinate buffer pH 5.0. The fusion protein was eluted in 100 mM
QB citrate buffer pH 3.0 and 50 mM NaCl. Immediately after elution,
1M Tris-HCl buffer pH 8.0 was added to the protein solution to
neutralize the pH. Protein aggregates were separated by size
exclusion chromatography (SEC) on Superdex 200 increase 10/300 GL
column (GE Healthcare Life Sciences). The SEC mobile phase buffer
was kept at 20 mM Tris-HCl pH 8.0, 100 mM NaCl and 50 mM arginine,
which was also the protein storage buffer. All chromatography steps
were performed on AKTA pure or AKTA Avant systems (GE Healthcare
Life Sciences).
[0335] Recombinant Expression and Purification of His-Tagged TREM2
ECD
[0336] The ecto domain (residues 19-172) of TREM2
(UniProtKB--Q9NZC2) was subcloned in the pRK vector with the
secretion signal from mouse Ig kappa chain V-III, amino acids 1-20
(UniProtKB ID--P01661) at the N-terminal region, and a 6.times.-His
tag (SEQ ID NO:35) at the C-terminal region. The insert was
verified by sequencing and maxi prep plasmid purification was
performed.
[0337] Purified plasmid was transfected into Expi293F.TM. cells
(Thermo Fisher) using the Expi293F.TM. Expression System Kit
according to the manufacturer's instructions. Transfected cells
were incubated in an orbital shaker (Infors HT Multitron) at 125
rpm and 37.degree. C. in a humidified atmosphere of 6% CO.sub.2.
ExpiFectamine.TM. 293 Transfection Enhancer 1 and 2 were added to
the cells 16 hours post transfection and the media supernatant was
harvested 96 hours post transfection.
[0338] Harvested media was supplemented with 1M imidazole pH 8.0 to
a final concentration of 10 mM and filtered using the Nalgene.TM.
Rapid-Flow.TM. disposable filter units (Thermo Fisher) with a pore
size of 0.4 microns. HisPur.TM. Ni-NTA Resin (Thermo Fisher) was
washed with MQ water and equilibrated with load buffer (20 mM Tris
pH 8.0, 150 mM NaCl, and 10 mM imidazole). Affinity purification
was performed using the gravity flow method. The harvested media
was loaded onto the resin and nonspecifically bound proteins were
washed with load buffer supplemented with 50 mM and 100 mM
imidazole. The bound His-tagged TREM2 eco domain was eluted with 20
mM Tris pH 8.0, 150 mM NaCl, and 200 mM imidazole. Eluted protein
was concentrated using Amicon 10 kDa concentrators and the
concentrated protein was further purified by gel filtration
chromatography using the AKTA Avant system (GE Healthcare Life
Sciences). The protein was loaded onto a HiLoad Superdex 200 16/600
(GE Healthcare Life Sciences) column equilibrated with 1.times.PBS
and eluted and fractionated using 1.times.PBS as the running
buffer. Eluted fractions were analyzed by electrophoresis on
polyacrylamide (PAGE) gels under denaturing and native conditions.
Eluted fractions were further characterized by analytical size
exclusion chromatography and the intact protein mass determination.
Results from the PAGE and analytical characterization were used to
pool the heavily glycosylated protein fractions and these were
aliquoted and stored at -80.degree. C.
[0339] Generation of Antibodies
[0340] Rodents (mice and rats) were immunized using standard
protocols with rhTREM2-Fc immunogen or BWZ cells expressing full
length Trem2 receptor. Titers were measured throughout immunization
using sera collected at different time points. The detection of an
antigen specific immune response was performed using flow cytometry
with the rhTREM2-Fc immunogen and live BWZ cells expressing
full-length TREM2. Selection criteria of candidate antibodies
included rodent antibody production and specificity of binding to
TREM2 as detected by flow cytometry. Antibody-secreting cells were
isolated from animal immune tissues including spleen, lymph nodes
and bone marrow.
[0341] Single cell suspensions were analyzed to determine the
binding properties of secreted antibodies. Antibody-secreting cells
were loaded into microfluidic devices and isolated in nanoliter
volume reaction chambers to enable the detection of secreted
antibodies using fluorescent and brightfield image-based microscopy
assays (see, e.g., U.S. Pat. No. 9,188,593). Binding assays
involving detection of antibodies binding to antigen-coated
micro-beads, detection of soluble fluorescently-labeled antigen
binding to antibodies immobilized on beads, and detection of
antibody binding to cell surface-expressed antigens were carried
out. Cell surface-expressed antigens included both recombinant form
and the native forms of antigens presented on the surface of
cells.
[0342] Image analysis was used to identify chambers exhibiting
positive fluorescent signals, indicating the presence of a single
cell producing antibodies with the desired properties, and the
contents of chambers were recovered and lysed in 384 well plates
(see, e.g., U.S. Pat. No. 10,087,408). Single cell lysates were
then subjected to RT-PCR to amplify the heavy and light chain
variable region sequences. The resulting amplicons were then
sequenced to determine the cDNA sequence of paired heavy and light
chain variable regions from the selected single cells. The
resulting sequences were manually inspected and analyzed to
determine sequence diversity and somatic hypermutation. Sequences
were selected for expression based on screening data and sequence
diversity. Expressed antibodies were tested to confirm antigen
binding specificity.
Example 2. Sequence Optimization and Humanization of Anti-TREM2
Antibodies
[0343] Exemplary anti-TREM2 antibodies were sequence optimized and
humanized, followed by characterization for binding kinetics and
binding specificity.
[0344] Sequence optimization was conducted by searching within CDR
sequences for residues that are susceptible to chemical
modification (e.g., asparagine deamidation motifs (NG), aspartic
acid isomerization motifs (DS), and potential oxidation residues
(tryptophan (W) and methionine (M)) and making amino acid
substitutions with conservative and germline residues to remove
such sequence liabilities. Humanized and sequence-optimized
variants of anti-TREM2 antibodies were then analyzed for binding
kinetics using Biacore and dose-titrated cell binding to HEK293-H6
cells (see, Example 5 for representative protocols).
Example 3. Generation of Anti-TREM2 Antibodies Having Modified Fc
Polypeptides ("ATV:TREM2")
[0345] The Fd (V.sub.H+CH1) region of a humanized, affinity matured
anti-TREM2 antibody (SEQ ID NOS:22 and 24) was cloned into
expression vectors comprising a sequence encoding an Fc polypeptide
engineered to bind to the human transferrin receptor (TfR)
(CH3C.35.23.1.1, CH3C.35.23.3, CH3C.35.23.3 cisLALA, or CH3C.35.24)
or a sequence encoding an Fc polypeptide that binds to the
cynomolgus monkey transferrin receptor (CH3C.35.21). The Fc
polypeptide-encoding sequence also contained a "knob" (T366W)
mutation to prevent homodimerization and promote heterodimerization
with an Fc polypeptide comprising "hole" (T366S/L368A/Y407V)
mutations. The Fd region was also cloned into corresponding "hole"
vectors comprising a sequence encoding an Fc polypeptide with hole
mutations, but lacking the TfR binding mutations. The coding
sequences (both Fd-knob-Fc and Fd-hole-Fc constructs) also
contained "LALA" (L234A; L235A) mutations in the hinge region to
reduce effector function (Wines et al., J. Immunol. 164:5313-5318
(2000) and "LS" (M428L; N434S) mutations in the Fc CH3 region to
increase binding to FcRn (see, e.g., Zalevsky et al., Nat. Biotech.
28(2):157-159 (2010)). The final encoded heavy chain sequences
expressed by the vectors are set forth in Table 1.
TABLE-US-00001 TABLE 1 ATV: TREM2 Sequences ATV: TREM2 First Heavy
Chain Second Heavy Chain #1 SEQ ID NO: 42 SEQ ID NO: 53 #2 SEQ ID
NO: 45 SEQ ID NO: 53 #3 SEQ ID NO: 48 SEQ ID NO: 53 #4 SEQ ID NO:
48 SEQ ID NO: 52 #5 SEQ ID NO: 51 SEQ ID NO: 53
[0346] The corresponding aforementioned knob and hole vectors were
co-transfected to ExpiCHO or Expi293 cells along with the
corresponding light chain vector (SEQ ID NO:54) in the ratio
knob:hole:light chain of 1:1:2. The expressed protein was purified
by Protein A chromatography followed by preparative size-exclusion
chromatography (SEC) to isolate purified anti-TREM2 protein.
[0347] Binding of anti-TREM2 protein to human transferrin receptor
was determined as follows: anti-human-Fab was immobilized on a CM5
chip, and the anti-TREM2 protein was captured. Full-length human
TfR or human TfR apical domain at serial dilution (e.g.,
concentrations of 1-1,000 nM) was flowed over the chip (180 second
association time) and then allowed to dissociate. Fitting was
performed using a 1:1 binding model.
Example 4. Characterization of Anti-TREM2 Antibodies
[0348] The following sections describe various assays that were
carried out to assess the binding and functional characteristics of
generated anti-TREM2 antibodies.
[0349] Affinity Measurement by Biacore Kinetic Measurement
[0350] Surface plasmon resonance (Biacore.TM. 8K instrument) was
used to measure anti-TREM2 antibody affinities for human and
cynomolgus TREM2 ECD. Anti-TREM2 antibodies were captured using
Human Fab Capture Kit (GE Healthcare Life Sciences, Catalog No.
28958325) on a Biacore Series S CM5 sensor chip (GE Healthcare Life
Sciences, Catalog No. 29149604). Serial 3-fold dilutions of
recombinant human or cynomolgus TREM2 were injected at a flow rate
of 30 .mu.L/min. Antibody binding was monitored for 300 seconds,
followed by monitoring of antibody dissociation for 600+ seconds in
HBS-EP+ running buffer (GE Healthcare Life Sciences, Catalog No.
BR100669). The binding response was corrected by subtracting the RU
value from a blank flow cell. A 1:1 Languir model of simultaneous
fitting of k.sub.on and k.sub.off was used for kinetics analysis.
K.sub.D binding values were calculated from k.sub.on and
k.sub.off.
[0351] Evaluation of TRFEM2 Binding in TREM2-Expressing HEK
Cells
[0352] The binding characteristics of anti-TREM2 antibodies was
evaluated in HEK 293 cells expressing human TREM2 as follows.
[0353] A HEK 293 cell line stably expressing human TREM2/DAP12 was
generated by transfecting the cells with a vector expressing wild
type human TREM2 and DAP12, and DAP12 alone, respectively. Stable
expressing clones were selected, and the cell surface TREM2
expression was evaluated by flow cytometry. APC-conjugated rat
anti-human/mouse-TREM2 monoclonal antibody (R&D, Catalog No.
MAB17291) was used to detect surface TREM2 expression. The clone
showing the highest wild type TREM2 expression level was selected
and named "HEK293-H6." The clones stably expressing DAP12 were
analyzed by Western blot, and the selected clone was named
"HEK293-DAP12#1."
[0354] HEK 293 overexpressing human TREM2 (HEK293-H6) and HEK 293
overexpressing GFP (B5) were harvested by 0.05% trypsin and
incubated at 37.degree. C. for 2 hours. After incubation, the cells
were centrifuged and washed in FACS buffer (PBS+0.5% BSA) twice.
Mixed cells were resuspended in FACS buffer with human Trustain FcX
solution (Biolegend, Catalog No. 422302) at a density of
10.sup.6/mL per cell line. The mixed cell lines were seeded at
200,000 cells per well in a 96-well round-bottom plate and
incubated for 20 minutes at room temperature. After incubation, the
cells were centrifuged and incubated with a dose titration of
anti-TREM2 antibodies for 45 minutes on ice. After incubation, the
cells were centrifuged and washed with FACS buffer three times. The
cells were then incubated with secondary antibody (Alexa Fluor 647
AffiniPure F(ab')2 Fragment Goat Anti-human IgG(H+L), Jackson
ImmunoResearch Laboratories, Catalog No. 109-606-088, 1:800
dilution) for 30 minutes on ice. After incubation, the cells were
washed with FACS buffer three times, resuspended in 100 .mu.L of
FACS buffer, and analyzed by flow cytometry (BD FACSCanto II, San
Jose, Calif.), for which 30,000 events were obtained for each
sample. Mean fluorescence intensity per cells were calculated by
FlowJo software and used for generating dose response binding
curve.
[0355] Activation of TREM2-Dependent pSyk Signaling
[0356] Activation of TREM2-dependent pSyk signaling was measured in
human macrophage cells or in HEK293-H6 cells using a commercial
AlphaLisa assay from Perkin-Elmer.
[0357] For all experiments involving use of lipid vesicles
containing 70% DOPC and 30% POPS, the lipid vesicles were prepared
within two weeks of experiments as follows: 7 mg DOPC
(1,2-dioleoyl-sn-glycero-3-phosphocholine) and 3 mg POPS
(1-palmitoyl-2-oleoyl-sn-glycero-3-phospho-L-serine) were combined
in chloroform in a glass vial and dried under a stream of N2 gas
for 1-2 hours, or until completely dry. The lipid mixture was
re-suspended in 1 mL HBSS (for a final lipid concentration of about
10 mg/mL) and vortexed for 2-3 minutes. Subsequently, the lipid
suspension was extruded using an Avanti mini-extruder constructed
with one 100-nm pore size membrane to form small unilamellar
vesicles at 10 mg/mL.
[0358] 1. Dosing of Antibodies in Cells
[0359] The day before assay, human macrophage cells or HEK293-H6
cells were plated at 100,000 cells/well or 40,000 cells/well,
respectively, on a 96-well plate coated with poly-D-lysine.
Antibodies were diluted in a 10-point serial dilution with 3-fold
dilution between points into PBS. For antagonist dose-response
curves, lipid vesicles containing 70% DOPC and 30% POPS at 1 mg/mL
final concentration were also included in the antibody/PBS mixture.
The cells were washed 3 times with HBSS using a Biotek 405/406
plate washer, after which 50 .mu.L per well of the antibody/PBS
(with or without vesicles) solution was added using a Hamilton
Nimbus liquid handler. The cell plate was then transferred to a
37.degree. C. incubator for 5 minutes. The liposome/antibody
solution was removed by flicking the plate, and 40 .mu.L lysis
buffer (Cell Signaling Technologies, CST) containing 1 .mu.M PMSF
was added using the liquid handler. The lysate was then either
frozen at -80.degree. C. or immediately assayed in the AlphaLisa
assay.
[0360] Human macrophage cells were prepared for assay as follows.
Human monocytes were isolated following the RosetteSep human
monocyte enrichment cocktail protocol (Stemcell Technologies, REF
#15068) from fresh blood. Isolated monocytes were washed in wash
buffer (PBS+2% FBS) and resuspended in 10 mL ACK lysis buffer
(ThermoFisher Scientific, Catalog No. A10492) to lyse red blood
cells. Twenty (20) mL of wash buffer was added to stop cell lysis,
and the sample was centrifuged and washed once more with culture
media (RPMI, 10% Hyclone FBS, 1% Sodium Pyruvate, 1% Glutamax, 1%
non-essential amino acids, and 1% Penicillin-streptomycin). Human
monocytes were then differentiated into macrophage cells in culture
media in the presence of 50 ng/mL human recombinant M-CSF (Gibco,
Catalog No. PHC9501) at 250-mL flask. Fresh human M-CSF was spiked
on day 3 and human macrophages were subsequently harvested on day 5
and used for assay.
[0361] 2. AlphaLisa Assay
[0362] Cell lysates were assayed for pSyk using the standard
protocol for the Perkin Elmer pSyk AlphaLisa kit. In brief, 10
.mu.L of lysate/well was transferred to a white opaque 384 well
Optiplate (Perkin Elmer). Next, 5 .mu.L of Acceptor Mix (containing
the working solution of acceptor beads) was added per well,
followed by sealing of plates with foil seals and incubation for 1
hour at room temperature. Subsequently, 5 .mu.L of Donor Mix
(containing the working solution of donor beads) was added to each
well under reduced light conditions. Plates were again sealed and
incubated for 1 hour at room temperature. Finally, the plates were
read using AlphaLisa settings on a Perkin Elmer EnVision plate
reader.
[0363] Survival Assay in Human Macrophage Cells
[0364] Human monocytes were isolated following the RosetteSep human
monocyte enrichment cocktail protocol (Stemcell Technologies,
Catalog No. 15068). Isolated monocytes were washed in wash buffer
(PBS+2% FBS) and resuspended in 10 mL ACK lysis solution
(ThermoFisher Scientific, Catalog No. A10492) to lyse red blood
cells. Twenty (20) mL wash buffer was added to stop lysis. The cell
suspension was centrifuged and washed once with culture media (RPMI
1640+10% FBS+penicillin/streptomycin). Cells were resuspended in
culture media at a density of 10.sup.6 cells .mu.L/mL and used in
the survival assay described below.
[0365] The day prior to assay, 96-well plates were pre-coated with
anti-TREM2 antibody or isotype control in a dose titration (45
.mu.L/well, total 12 points) and incubated overnight at 4.degree.
C. After overnight incubation, the pre-coated plate was washed
twice with PBS and then loaded with human monocyte (10.sup.5
cells/well) in the presence of low concentration human M-CSF (5
ng/mL, Gibco, Catalog No. PHC9501). After 5 days at 37.degree. C.,
the media was aspirated, and 100 .mu.L PBS+100 .mu.L Celltiter-glo
media (Promega, Catalog No. G7571) was added to each well. After 10
minutes of incubation, the cell media was transferred to multiwell
plates compatible for luminometer use, and luminescence for cell
viability was recorded.
[0366] Lipid Storage Assay
[0367] Prior to assay, induced human pluripotent stem cells (iPSCs)
were first differentiated into hematopoietic progenitor cells
(HPCs) using a commercially available kit (STEMdiff Hematopoietic
Kit from StemCell Technologies). HPCs were transferred to a plate
containing primary human astrocytes and co-cultured for 14-21 days.
Once floating cells in co-culture were predominantly identified as
mature microglia (>80%), the microglia were used for assay.
[0368] Cells (iPSC-derived human microglia, 30,000 cells/well) were
plated on PDL-coated 96-well plates in full serum media. After 24
hours at 37.degree. C., the media was exchanged for full serum
media containing oleic acid-albumin (10 .mu.M or 33 .mu.M final
concentration, Sigma O3008) or purified unlabeled myelin (50
.mu.g/mL final concentration, purified from wildtype C57Bl/6 mouse
brain (Jackson Laboratories) using methods described in Safaiyan et
al. (2016, Nature Neuroscience 19(8):995-998)). After 24 hours at
37.degree. C. of lipid treatment, the media was exchanged for media
containing anti-TREM2 antibody. For single point experiments, the
concentration of anti-TREM2-antibody used was 100 nM. For
dose-response curves, media containing 100 nM anti-TREM2 antibody
was serially diluted 3-fold for a total of 10 points. RSV was used
as a control. The cells were incubated for another 48 hours at
37.degree. C. before imaging cells using Bodipy stain, or
extracting the cells for lipidomics, as described below.
[0369] For Bodipy imaging, the supernatant was removed, and cells
were incubated at 37.degree. C. for 30 minutes in live cell imaging
buffer (Life Technologies, Catalog No. A14291DJ) containing 1:2500
of a 1 mg/mL Bodipy 493/503 solution in DMSO (Thermo-Fisher D3922)
and 1 drop/mL of Nucblue (ThermoFisher, Catalog No. R37605). After
the incubation period, the staining solution was removed, and the
cells were either imaged live or fixed in 4% paraformaldehyde. The
cells were imaged using the Alexa 488 channel for Bodipy, and DAPI
illumination settings on an Opera Phoenix high content confocal
imager. Lipid spots were analyzed using a spot-finding algorithm on
the Harmony software supplied with the instrument.
[0370] For lipidomic analysis, cells were washed once with PBS
while kept on ice. A volume of 70 .mu.L of a 9:1 methanol:water
solution containing 1:100 internal standards was added to the cells
in the 96-well plate. The plate was agitated on a shaker at
4.degree. C. and 1200 rpm for 20 minutes and then centrifuged for 5
minutes at 300.times.g. A 50 .mu.L sample of supernatant was
transferred to LCMS vials and kept at -80.degree. C. until analyzed
on the instrument.
[0371] Lipid levels were analyzed by liquid chromatography
(Shimadzu Nexera X2 system, Shimadzu Scientific Instrument,
Columbia, Md., USA) coupled to electrospray mass spectrometry
(QTRAP 6500+, Sciex, Framingham, Mass., USA). For each analysis, 5
.mu.L of sample was injected on a BEH C18 1.7 .mu.m, 2.1.times.100
mm column (Waters Corporation, Milford, Mass., USA) using a flow
rate of 0.25 mL/min at 55.degree. C. For positive ionization mode,
mobile phase A consisted of 60:40 acetonitrile/water (v/v) with 10
mM ammonium formate+0.1% formic acid; mobile phase B consisted of
90:10 isopropyl alcohol/acetonitrile (v/v) with 10 mM ammonium
formate+0.1% formic acid. For negative ionization mode, mobile
phase A consisted of 60:40 acetonitrile/water (v/v) with 10 mM
ammonium acetate; mobile phase B consisted of 90:10 isopropyl
alcohol/acetonitrile (v/v) with 10 mM ammonium acetate. The
gradient was programmed as follows: 0.0-8.0 min from 45% B to 99%
B, 8.0-9.0 min at 99% B, 9.0-9.1 min to 45% B, and 9.1-10.0 min at
45% B. Electrospray ionization was performed in either positive or
negative ion mode applying the following settings: curtain gas at
30; collision gas set at medium; ion spray voltage at 5500
(positive mode) or 4500 (negative mode); temperature at 250.degree.
C. (positive mode) or 600.degree. C. (negative mode); ion source
Gas 1 at 50; ion source Gas 2 at 60. Data acquisition was performed
using Analyst 1.6.3 (Sciex) in multiple reaction monitoring mode
(MRM), with the following parameters: dwell time (msec) and
collision energy (CE); declustering potential (DP) at 80; entrance
potential (EP) at 10 (positive mode) or -10 (negative mode), and
collision cell exit potential (CXP) at 12.5 (positive mode) or
-12.5 (negative mode). Lipids were quantified using a mixture of
non-endogenous internal standards. Lipids were identified based on
their retention times and MRM properties of commercially available
reference standards (Avanti Polar Lipids, Birmingham, Ala.,
USA).
[0372] Activation of TREM2-Dependent mTOR Signaling
[0373] Wild-type iPSC-derived human microglia were cultured and
treated with anti-TREM2 antibody (100 nM final concentration) and
either DMSO or a commercial mTOR inhibitor (Selleckchem, Catalog
No. AZD8055, 20 nM final concentration) for 96 hours. The treated
cells were subsequently lysed, and the cell lysates were prepared
for Western blots to investigate phosphorylation of major signaling
targets in the mTOR pathway. Primary antibodies for Western blots
were obtained from Cell Signaling Technologies: (1) phospho-mTOR
(Ser2448), Product No. 5536T; (2) mTOR (7C10), Product No. 2983T;
(3) phospho-AKT (Ser473), Product No. 9271T; (3) phospho-GSK-3beta
(Ser9), Product No. 5558T; (4) phospho-S6 ribosomal protein
(S235/236), Product No. 4858T; (5) phospho-4E-BP1 (Thr37/46),
Product No. 2855T; (6) beta-actin, Product No. 58169S.
Example 5. Results
[0374] Results for an analysis of the binding characteristics of
humanized and sequence-optimized variants of antibody CL0020188 are
provided in Table 2 and FIGS. 1A-1H. NG motifs in the CL0020188
CDR-H2 sequence (SEQ ID NO:5) and CDR-L1 sequence (SEQ ID NO:7)
were modified, grafted onto human framework regions, and analyzed.
Table 2 provides K.sub.D values as measured by Biacore and
EC.sub.50 values as measured by dose-titrated binding assay in
HEK293-H6 cells. FIGS. 1A-1H include representative dose-response
curves of binding to TREM2 expressed by HEK293-H6 cells for the
humanized and sequence-optimized variants. Variants are represented
by solid black circles (.circle-solid.), while isotype controls are
represented by open white circles (.smallcircle.).
TABLE-US-00002 TABLE 2 Binding Characteristics of
Sequence-Optimized and Humanized Variants of CL0020188 Clone
hV.sub.H hV.sub.L K.sub.D EC.sub.50 CL0020188-1 NG/graft NG/graft
2.3 nM 0.42 nM CL0020188-2 NG/3m NG/graft 3.4 nM 0.26 nM
CL0020188-3 NG/graft TG/graft 6.8 nM 0.64 nM CL0020188-4 NG/3m
TG/graft 4.8 nM 0.44 nM CL0020188-5 NA/graft NG/graft 5.1 nM 0.45
nM CL0020188-6 NA/3m NG/graft 4.0 nM 0.31 nM CL0020188-7 NA/graft
TG/graft 10 nM 0.68 nM CL0020188-8 NA/3m TG/graft 7.3 nM 0.51 nM
Parent 9.5 nM 0.44 nM 3m = A24G/L45P/V48L in V.sub.H
[0375] As illustrated in Table 2, humanized and sequence-optimized
clones of CL00201088 exhibited similar affinity values for hTREM2
compared to the parent antibody (K.sub.D=9.5 nM), as measured by
Biacore. This was consistent with cell-binding results in HEK293-H6
cells, which are illustrated in Table 1, with corresponding
dose-response curves provided in FIGS. 1A-1H. Compared to the
parent antibody (EC.sub.50=0.44 nM), humanized and
sequence-optimized clones exhibited comparable and sub-nanomolar
affinity for TREM2 expressed in HEK293-H6 cells. Taken together,
the results indicate comparable binding kinetics between the parent
antibody and the humanized and sequence-optimized variants.
[0376] Results for ATV:TREM2 variants described in Example 3 are
summarized in Table 3 below. An exemplary cell binding curve based
on binding to human TREM2-expressing HEK cells and analysis by FACS
is illustrated in FIG. 2.
TABLE-US-00003 TABLE 3 Summary of ATV: TREM2 Characteristics EC50
(nM) ATV: Biacore K.sub.D (nM) Human TREM2 TREM2 Human TREM2 Cyno
TREM2 Human TfR HEK #1 TBD TBD TBD TBD #2 5.4 2.4 650 4.9 #3 3.0
2.2 1400 4.6 #4 2.0 2.2 TBD TBD #5 5.7 2.4 620 3.6 TBD = to be
determined
[0377] The antibodies were also assessed for TREM2-dependent pSyk
signaling in HEK-H6 cells, capability for promoting survival of
human macrophage cells, and ability to modulate lipid accumulation
in iPSC-derived human microglial cells (hereinafter referenced as
"iPSC microglia" or "iMG"). FIG. 3 illustrates the results for an
ATV:TREM2 variant (ATV:TREM2 #3) and a corresponding anti-TREM2
antibody. The ATV:TREM2 was able to activate pSyk signaling in
TREM2-expressing HEK293-H6 cells to a significantly greater extent
than the corresponding TREM2 antibody, indicating that the addition
of ATV to the molecule can increase its potency (FIG. 3). In
addition, the ATV:TREM2 induced macrophage survival with an
EC.sub.50 of 4.1+0.3 nM. Finally, the anti-TREM2 antibodies
demonstrated capability in reducing lipid accumulation in
myelin-treated iMG (FIGS. 4A and 4B) with an IC.sub.50 for
inhibition of lipid storage of 0.20 nM (97.7+0.3% max.
inhibition).
[0378] Additional studies were carried out to investigate the
ability of ATV:TREM2 to reduce lipid accumulation. FIGS. 5A-5F and
FIGS. 6A-6C show that a representative ATV:TREM2 variant (ATV:TREM2
#3) reduces lipid accumulation while enhancing fatty acid oxidation
intermediates, suggesting a potential role of ATV:TREM2 in
enhancing mitochondrial function. Cells (iMG) treated with oleic
acid lipid challenge (33 .mu.M) followed by incubation with
ATV:TREM2 were able to reduce lipid accumulation, as illustrated by
Bodipy staining (FIGS. 5A and 5B). LCMS analysis of iMG treated
with myelin for 24 hours, followed by incubation with ATV:TREM2 for
48 hours, indicated that ATV:TREM2 reduces triglyceride (TG)
species while concomitantly increasing beta-oxidation intermediates
(acyl carnitines) and TCA cycle intermediates (FIGS. 5C-5F). FIG.
5C provides a heat map showing all TG, acyl carnitine, and TCA
cycle intermediate species that illustrated a fold change of
>1.5 (p<0.05), while FIGS. 5D-5F illustrate the changes of
representative species in vehicle and myelin-challenged iMG that
were incubated with ATV:TREM2 or isotype control following
challenge. FIGS. 6A-6C illustrate the changes of specific TG, acyl
carnitine, and TCA cycle intermediate species in iMG incubated with
ATV:TREM2 or isotype control following myelin challenge. As
depicted in FIGS. 6A-6C, ATV:TREM2 reduces all species of TG and
ceramides while increasing certain species of short chain acyl
carnitines, indicating that ATV:TREM2 may enhance mitochondrial
function.
[0379] Microglial cell proliferation is associated with mTOR signal
activation and engagement. The role of ATV:TREM2 in downstream mTOR
pathway signaling was thus explored. The phosphorylation status of
mTOR signal pathway targets were analyzed by Western blot in
wild-type iPSC microglia incubated with a representative ATV:TREM2
variant (ATV:TREM2 #3) in the presence and absence of an mTOR
inhibitor. FIG. 7A illustrates representative Western blot images
of mTOR signal pathway targets. Quantification of Western blot data
is provided in FIGS. 7B-7E (phosphorylation levels normalized to
beta-actin loading control). Phosphorylation levels for each
ATV:TREM2-treated sample were compared to isotype antibody
control-treated samples for each independent experiment (n=6). The
results show that ATV:TREM2 activates mTOR pathway signaling, as
evidenced by increased phosphorylation levels of mTOR at
serine.sup.2488, AKT at serine.sup.473, ribosomal protein S6 (RPS6)
at serine.sup.235/236, and GSK3b at serine.sup.9 in
ATV:TREM2-treated samples relative to isotype controls (FIGS.
7B-7E; statistical summary: "ns" (p>0.05); "*" (p<0.01); "**"
(p<0.001)). RPS6 is a signal target that is downstream of mTORC1
complex, and GSK3b is a signal target that is downstream of mTORC2
complex. For all data generated in FIGS. 5A to 7E, the isotype
control ("ISO") for ATV:TREM2 #3 contains the sequences provided in
Table 5.
Example 6. ATV:TREM2 Role in Lysosomal Dysfunction
[0380] The role of ATV:TREM2 in lysosomal function was
investigated. To assess impact on lysosomal function, levels of
progranulin (PGRN) and bis(monoacylglycero)phosphates (BMPs) were
measured in iPSC-derived microglial cells ("iMG") treated with an
ATV:TREM2 variant.
[0381] To evaluate the effect of ATV:TREM2 on PGRN levels, iMG were
plated at a density of 30,000 cells per well in a 96-well plate,
and incubated with ATV:TREM2 #3 (100 nM) for 72 hours, after which
the cell supernatant and cell lysate were collected and analyzed
for progranulin (PGRN) levels using a colorimetric sandwich
ELISA.
[0382] For measurement of PGRN levels, Thermo Scientific 384-well
Maxisorp plates were coated with 4 .mu.g/mL capture antibody
(R&D anti-PGRN antibody from DuoSet ELISA kit, Catalog No.
DY2420) diluted in phosphate-buffered saline (PBS) and incubated
overnight at 4.degree. C. The sample wells were blocked for 90
minutes with 3% BSA in PBS. Cell samples were diluted 1:10 in 3%
BSA in PBS and added to each sample well on the plate, followed by
incubation for 90 minutes at room temperature. Detection antibody
(R&D anti-PGRN antibody from DuoSet ELISA kit, Catalog No.
DY2420) diluted to 125 ng/mL was subsequently added to each sample
well, and the plate was incubated for 90 minutes at room
temperature. Lastly, HRP-conjugated Streptavidin (R&D SA-HRP
from DuoSet ELISA kit, Catalog No. DY2420) was diluted 1:200 and
added to each sample well. The plate was incubated for 20 minutes
at room temperature. After washing the sample wells with PBS,
development reagent (TMB substrate) was added and allowed to react
for 5 minutes before the reaction was stopped with 4 N
H.sub.2SO.sub.4. Absorbance was measured using a BioTek Synergy
Neo2 plate reader, and PGRN levels were determined by interpolation
from the standard curve fit with a four-parameter logistic curve.
As illustrated in FIG. 8, incubation with ATV:TREM2 increased PGRN
levels in both the supernatant and cell lysate relative to isotype
control.
[0383] To assess effect on BMP levels, iMG were challenged with
myelin or vehicle for 24 hours, followed by incubation with
ATV:TREM2 #3 (100 nM) for 48 hours. Cellular lipids were extracted
via addition of methanol containing an internal standard mixture
and BMP abundance was measured by liquid chromatography-mass
spectrometry (LC-MS/MS) on a Q-trap 6500 (SCIEX) similar to that
described in International PCT Publication No. WO 2020/112889. BMP
species were quantified using BMP(14:0_14:0) as the internal
standard and identified based on their retention times and MRM
properties. Quantification was performed using MultiQuant 3.02
(Sciex) after correction for isotopic overlap. BMP species were
normalized to median lipid content of all species measured. Protein
concentration was measured using the bicinchoninic acid (BCA) assay
(Pierce, Rockford, Ill., USA). A representative BMP result is
illustrated in FIG. 9. As shown in FIG. 9, iMG challenged with
myelin and subsequently treated with ATV:TREM2 reduces levels of
BMP species, suggesting potential rescue of lysosomal challenge
induced by myelin.
[0384] As illustrated in FIGS. 8 and 9, incubation of iMG with
ATV:TREM2 increases PGRN levels and corrects myelin-induced BMP
levels, indicating a role for ATV:TREM2 in modulating lysosomal
effects. In FIGS. 8 and 9, the isotype control (ISO) for ATV:TREM2
#3 contains the sequences provided in Table 5.
Example 7. ATV:TREM2 Role in Mitochondrial Respiration
[0385] To assess impact on mitochondrial respiration, oxygen
consumption was measured in iPSC-derived microglial cells ("iMG")
treated with an ATV:TREM2 variant or isotype control using a
Seahorse XFe96 analyzer (Agilent) and using materials and protocol
from the Seahorse XF Palmitate Oxidation Stress Kit (Agilent
103693). Cells were cultured on XF96 microplates (Agilent, Cat. No.
102416) with ATV:TREM2 #3 (100 nM) or isotype control for 72 hours
prior to the Seahorse experiment. Cells were subjected to
substrate-limiting media composed of Seahorse XF RPMI (Agilent
103576) supplemented with 0.5 mM glucose (Agilent 103577), 1 mM
glutamine (Agilent 103579), 0.5 mM L-carnitine (part of the
Seahorse XF Palmitate Oxidation Stress Kit), and 1% Hyclone FBS 16
hours prior to Seahorse experiment. Cells were dosed with either
palmitate-BSA conjugate (166 .mu.M) or BSA control immediately
before the experiment. Sequential injection of (1) etomoxir (4
.mu.M) (carnitine palmitoyltransferase 1 (CPT1) inhibitor) or
vehicle, (2) oligomycin (1.5 .mu.M) (ATP synthase complex V
inhibitor), (3) Carbonyl cyanide
4-(trifluoromethoxy)phenylhydrazone (FCCP, a mitochondrial
decoupling agent, 1 .mu.M), and (4) rotenone/antimycin (0.5 mM
each, complex I and complex III inhibitors, respectively) were used
to evaluate the mitochondrial respiration capacity of the cells.
Oxygen consumption rate was measured during the course of the
experiment by Seahorse analyzer (Agilent). A summary of
experimental conditions for evaluating mitochondrial respiration is
provided in Table 4.
TABLE-US-00004 TABLE 4 Experimental Conditions Condition Antibody
CPT1 Inhibitor Recovery substrate 1 Isotype Control Etoxomir BSA
(control) 2 ATV: TREM2 Vehicle BSA (control)) 3 Isotype Control
Etoxomir Palmitic acid (PAL) 4 ATV: TREM2 Vehicle Palmitic acid
(PAL) 5 Isotype Control Etoxomir BSA (control) 6 ATV: TREM2 Vehicle
BSA (control)) 7 Isotype Control Etoxomir Palmitic acid (PAL) 8
ATV: TREM2 Vehicle Palmitic acid (PAL)
[0386] A representative kinetic graph of oxygen consumption is
illustrated in FIG. 10A, and a bar graph illustrating the maximal
respiratory capacity of the cells is provided in FIG. 10B. As shown
in the figures, ATV:TREM2 increases maximal respiration to an
extent similar to that of fatty acid substrate palmitic acid (PAL).
This effect is diminished in the presence of CPT1 inhibitor. The
results indicate that ATV:TREM2 enhances maximal mitochondrial
respiration and that this effect appears to be conferred by
enhanced fatty acid oxidation capacity.
Example 8. ATV:TREM2 Comparative Properties
[0387] The properties of ATV:TREM2 #1 and ATV:TREM2 #3 were
compared to those of reference antibodies that bind TREM2, which
are described in WO 2019/028292. The heavy chain and light chain
sequences of reference antibody #1 ("Ref. Ab. #1") are represented
by SEQ ID NOs:74 and 75, respectively. The heavy chain and light
chain sequences of reference antibody #2 ("Ref. Ab. #2") are
represented by SEQ ID NOs:76 and 75, respectively. Heavy and light
chain sequences for the isotype controls of each anti-TREM2
antibody are provided in Table 5.
TABLE-US-00005 TABLE 5 Isotype Control Sequences Isotype Control
First Heavy Chain Second Heavy Chain Light Chain ISO for reference
SEQ ID NO: 77 SEQ ID NO: 77 SEQ ID NO: 78 antibody #1 and #2 ISO
for ATV: TREM2 #1 SEQ ID NO: 79 SEQ ID NO: 81 SEQ ID NO: 82 ISO for
ATV: TREM2 #3 SEQ ID NO: 80 SEQ ID NO: 81 SEQ ID NO: 82
[0388] ATV:TRFEM2 is More Potent In Vitro and Induces Less
Inflammation (Less Cytokine Release) than Reference Abs
[0389] Anti-TREM2 antibodies were evaluated using the human
macrophage cell survival assay described in Example 4. FIG. 11A
illustrates cell viability dose-response curves with the anti-TREM2
antibodies, with "ISO" referring to the isotype control for
ATV:TREM2 #3 (Table 5). Corresponding potency (EC50) and maximum
response (Emax) values as determined from the dose-response curves
are provided in FIGS. 11B and 11C; individual marks represent
single values from human cell donors (n=3). The results provided in
FIGS. 11A-11C indicate that ATV:TREM2 #3 is more potent in
promoting human macrophage survival in vitro than reference
antibodies #1 and #2.
[0390] In a separate experiment, human macrophage cells were
treated with 100 nM of surface-immobilized anti-TREM2 antibodies
for five days, after which cell culture media for each set of cells
was collected and analyzed for cytokine release using Luminex xMAP
technology and a commercial bead-based multiplex assay kit (Human
Cytokine 42-Plex Discovery Assay.RTM., Eve Technologies Corp.). A
heat map of relative cytokine release level is illustrated in FIG.
12 (Z-score used for plotting). The results in FIG. 12 illustrates
that ATV:TREM2 #3 induces less inflammation in human macrophage
cells than reference antibodies #1 and #2.
[0391] In summary, ATV:TREM2 #3, reference antibody #1, and
reference antibody #2 are able to promote human macrophage survival
and proliferation (FIG. 11A). However, ATV:TREM2 #3 shows stronger
potency for cell survival with reduced overall cytokine signature
relative to reference antibodies #1 and #2 (FIGS. 11B, 11C, and
12).
[0392] ATV:TRFEM2 is Able to Reduce Triglyceride Species Levels
after Challenge
[0393] Anti-TREM2 antibodies were evaluated by lipid storage assay
(using 10 .mu.M oleic acid challenge) as described in Example 4.
The results for ATV:TREM2 #3, reference antibody #1, and reference
antibody #2 are provided in FIGS. 13A-13E.
[0394] FIGS. 13A-13C illustrate volcano plots with cut-offs of
p<0.05 and fold change >1.5 for triglyceride species in
iPSC-derived microglial cells ("iMG") as quantified by LCMS. The
data is normalized to the isotype control for each antibody. As
shown in FIGS. 13A-13C, ATV:TREM2 #3 is able to modulate
triglyceride (TG) species post-oleic acid dosing, while the
reference antibodies did not significantly change levels of TG
species after oleic acid challenge. FIGS. 13D and 13E illustrate
bar graphs of representative TG species measurement, with data
normalized to isotype control for ATV:TREM2 #3.
[0395] ATV:TRFEM2 is Able Modulate TREM2 Levels In Vitro
[0396] To assess how anti-TREM2 antibodies impact TREM2 levels in
iPSC-derived microglial cells ("iMG"), iMG were incubated with
ATV:TREM2 #3 or reference antibody #1 for 72 hours at various
concentrations, followed by measurement of TREM2 levels in iMG cell
lysate and cell culture medium. TREM2 was measured as follows.
Briefly, MSD small spot streptavidin plates (Meso Scale Discovery)
were coated with biotinylated goat anti-hTREM2 polyclonal antibody
(R&D Systems, BAF1828) at room temperature for 1 hour. The
plates were then blocked with MSD Block A buffer (Meso Scale
Discovery) for 1 hour at room temperature. Samples and standards
were prepared/diluted with assay buffer (25% MSD Block A buffer in
TBST), and 30 .mu.L of samples and standards was loaded into the
plate after blocking. After 1 hour of incubation at room
temperature, the plates were washed with TBST and followed by
binding the primary antibody (ATV:TREM2 #3) for 1-hour at room
temperature. Afterwards, diluted sulfo-tagged goat anti-human IgG
(Southern Biotech, 2049-01) was added to the plates, and incubated
for one hour at room temperature. After washing with TBST, the MSD
plates were developed using 2.times.MSD read buffer T, followed by
detection using an MSD Sector plate reader. MSD values were
converted to absolute quantities of TREM2 by fitting a standard
curve using Meso Scale Discovery software (Discovery
Workbench).
[0397] FIGS. 14A and 14B show plots of TREM2 levels as a function
of antibody concentration in iMG cell lysates and cell culture
media after incubation with anti-TREM2 antibodies. TREM2 levels for
each antibody are normalized to their specific isotype control. In
FIGS. 14A and 14B, "ISO" represents isotype control for ATV:TREM2
#3. As illustrated in FIG. 14A, the levels of total TREM2 increased
with increasing amounts of antibody in the cell lysates of iMG
treated with ATV:TREM2 #3 and reference antibody #1. In contrast,
the levels of soluble TREM2 decreased with increasing amounts of
antibody in the cell culture media of the antibody-treated
cells.
[0398] ATV:TREM2 Exhibits a Superior Pharmokinetic Profile in
Non-Human Primates
[0399] The pharmacokinetic (PK) properties of anti-TREM2 antibodies
were studied in non-human primates. Briefly, young adult/adult male
cynomolgus monkeys ranging in age from 36 to 53 months were
intravenously administered a single dose of 30 mg/kg test article
(Table 6, n=5 per cohort). CSF samples were collected at 24, 168,
and 336 hours post-dose.
[0400] The results are illustrated in Table 6 and FIG. 15. Table 6
shows the PK values of anti-TREM2 antibodies in the cerebrospinal
fluid (CSF) of non-human primates. The PK values of ATV:TREM2
variants show that these antibodies have a higher exposure in CSF
relative to TREM2 Ab (lacking binding capacity for transferrin
receptor; heavy and light chain sequences represented by SEQ ID
NOs: 83 and 54, respectively) and reference antibody #1.
TABLE-US-00006 TABLE 6 CSF pharmacokinetic profiles of anti-TREM2
antibodies in non-human primates Test Article Dose (mg/kg)
C.sub.max (nM) T.sub.max (hr) AUC.sub.0-last (nM * hr) ATV: TREM2
#1 30 3.17 24 822.59 ATV: TREM2 #3 30 1.49 24 461.79 ISO* 30 3.23
24 618.90 TREM2 Ab 30 0.98 24 280.38 Reference 30 0.70 24 166.69
antibody #1 *Isotype control for ATV: TREM2 #1
[0401] FIG. 15 illustrates the PK profiles for anti-TREM2
antibodies in the CSF of dosed non-human primates. As illustrated
in FIG. 15 and Table 6, ATV:TREM2 shows at least a 2-fold increase
in CSF PK compared to reference antibody #1.
Example 9. ATV:TREM2 Properties in a Humanized TREM2 Mouse
[0402] To investigate the effects of ATV:TREM2 in vivo, we
generated a BAC transgenic mouse that expresses human TREM2. These
mice were crossed to a human transferrin receptor knock-in mouse
described in U.S. Pat. No. 10,143,187 to allow for characterization
of the ATV:TREM2 molecules described herein.
[0403] Generation of Human TREM2 BAC Transgenic Model
[0404] To investigate the effects of ATV:TREM2 in vivo, we
generated several transgenic mouse lines expressing the human TREM2
by using modified BAC DNA CTD-2210D2 (ThermoFisher Scientific; Cat.
No. 96012). In the original BAC DNA CTD-2210D2, the human TREM2
coding region and its regulatory elements are flanked by two other
TREM-like genes, TREML1 and TREML2. To avoid interference from
these TREM-like genes, we abolished their expression by deleting
exon 1 from TREML1 and exon 3 from TREML2. This engineered BAC
CTD-2210D2 DNA construct was injected into the pronucleus of
fertilized mouse eggs from C57BL/6J mice. Two independent founder
lines (termed TB36 and TB45) were obtained and were shown to have
germline transmission.
[0405] To characterize and compare these two transgenic lines,
hemizygous transgenic animals and wild type non-transgenic litter
mate controls were used for analysis. Human TREM2 copy number was
determined using qPCR analysis of tail genomic DNA. Human TREM2
mRNA and protein levels were measured in brains, liver, lung, and
spleen by qRT-PCR and an MSD assay. Human TREML1 and TREML2 mRNA
were analyzed in brain-sorted microglia by qRT-PCR. Surface TREM2
expression was quantified by FACS in bone marrow derived
macrophages (BMDM). Human TREM2 function was assessed by the in
vitro BMDM survival assay.
[0406] Human TREML1 and TREML2 were undetectable in either TB36 or
TB45, showing successful deletion of these genes. qPCR analysis
showed that there are two and one copies of human TREM2 transgenes
in TB36 and TB45, respectively, with corresponding higher human
TREM2 expression in TB36 relative to TB45 in brains and peripheral
tissues. The in vitro survival assay showed that human TREM2
agonist antibodies trigger stronger responses in TB36 line than in
TB45 line.
[0407] Based on this ex vivo and in vitro characterization, we
selected TB36 for the following breeding and in vivo studies. The
hemizygous TB36 mice were further backcrossed to C57BL/6J for three
rounds and then bred with hTfR KI mice (described in U.S. Pat. No.
10,143,187) to generate human TREM2 BAC hemizygous; hTfR KI
homozygous mice for in vivo studies.
[0408] Pharmacokinetics and Pharmacodynamics Responses with ATV:
TREM2 in TB36 hT R KI Mice
[0409] To determine whether ATV:TREM2 can trigger microglia
response in vivo, a single dose of ATV:TREM2 #3 or a corresponding
isotype control (ATV:RSV) (100 mg/kg) was intravenously
administered to TB36/hTfR KI mice at day 0, and mice were
sacrificed at day 1 or day 4 post-dose for ex vivo analysis. At the
time of sacrifice, animals were anesthetized by intraperitoneal
injection of 2.5% Avertin. Terminal blood was collected through the
cardiac puncture in an EDTA tube with slow inversion (10 times) and
was then centrifuged at 15,350 g for 7 minutes at 4.degree. C.
Plasma (top layer) was transferred to a 1.5-ml Eppendorf tube and
stored at -80.degree. C. until measurement. After blood collection,
CSF samples were collected by pre-pulled glass capillary tubes from
the cisterna magna and then transferred to 0.5 mL Protein LoBind
Eppendorf tubes for centrifugation at 12,700 rpm for 7 minutes at
4.degree. C. The supernatants of CSF samples were snap frozen on
dry ice and stored at -80.degree. C. until measurement. The animals
were then perfused with cold PBS, and brains were dissected out,
and the two hemispheres were separated. Right hemi-brains were
immersion fixed in 4% paraformaldehyde at 4.degree. C. for 24 hours
and then transferred to a phosphate buffered saline (PBS) solution
with 0.1% sodium azide for storage until ready for 30% sucrose
processing and sectioning. Left hemi-brains were cut into two
pieces and snap frozen in two tubes for the PK measurement and
other target engagement/cytokines analyses, respectively.
[0410] Plasma and brain levels of human IgG were evaluated at 1 day
and 4 days post-dose and are shown in Table 7 below.
TABLE-US-00007 TABLE 7 Plasma and Brain pharmacokinetic profiles of
in TB36/hTfR-KI mice Plasma Brain TB36 * ATV: RSV ATV: 188-14 ATV:
RSV [hIgG] ATV: 188-14 hTfR-KI [hIgG] uM [hIgG] uM nM [hIgG] nM 24
hours 2.96 .+-. 0.17 1.78 .+-. 0.073 34.1 .+-. 3.2 24.4 .+-. 1.6 96
hours 0.401 .+-. 0.074 0.29 .+-. 0.013 13.8 .+-. 1.8 6.67 .+-.
1.0
[0411] Effect of ATV:TREM2 on Microglial Proliferation
[0412] Four doses of 5-Ethynyl-2'-deoxyuridine (EdU, 80 mg/kg), a
thymidine analog that can be incorporated into newly synthesized
DNA, were intraperitoneally administered to the mice at day 0, day
1, day 2 and day 3 after the treatment of ATV:TREM2 #3 or ATV:RSV
at day 0.
[0413] For detection of proliferating microglia (EdU+Iba1+), the
brain sections at day 4 post-dose were treated with Click-iT
EdU-imaging kits (ThermoFisher Scientific, C10637) followed by Iba1
immunostaining. EdU-Iba1 staining showed that ATV:TREM2
dramatically increases newborn microglia number (EdU+ Iba1+; FIG.
16A) and total microglial coverage area (Iba1+ area; FIG. 16B) in
the brains compared to the isotype control, thus demonstrating that
ATV:TREM2 significantly increases microglial proliferation as
compared to the control.
[0414] ATV:TREM2 is Dramatically More Potent for Microglial
Proliferation, Compared to Non-ATV TREM2 Antibodies in Mouse
Model
[0415] To determine the lowest effective dosage of ATV:TREM2 and to
compare to non-ATV TREM2 antibodies, a single dose of ATV:TREM2 #3
(1, 3, 10, 30 mg/kg) or a corresponding TREM2 reference antibody
("TREM2 Ab;" 30 mg/kg), reference antibody #2 (30 mg/kg) or isotype
control (ATV:RSV; 30 mg/kg) were intravenously administered to
TB36/hTfR KI mice at day 0. To measure microglial proliferation,
mice were IP administrated with EdU at day 0, day 1, day 2 and day
3 after antibody treatment. Mice were taken down at day 1 and day 4
for analysis. EdU-Iba1 double staining shows dosage-dependent
increase of new born microglia from 1 mg/kg to 10 mg/kg of the
ATV:TREM2-dosed animals at day 4, with 10 mg/kg ATV:TREM2 showing
the maximum effect on microglial proliferation (FIGS. 17A and 17B).
Furthermore, the effect of 1 mg/kg ATV:TREM2 on microglial
proliferation is slightly higher (not statistically significant)
than either anti-TREM2 or reference antibody #2, despite 30-fold
higher dosing.
[0416] ATV:TREM2 Transiently Increases Brain Cytokine Levels in
TB36/hTfR KI Mice
[0417] To determine the effect of ATV:TREM2 on cytokine (e.g.,
chemokine) levels, cytokine levels were measured in terminal plasma
and brain lysates (prepared by Cell Signaling lysis buffer #9803)
of TB36/hTfR KI mice using the Mouse Cytokine Array/Chemokine Array
44-Plex (MD44). We found that treatment with ATV:TREM2 #3 did not
change the cytokine levels measured in the plasma, but acutely
increased some cytokine levels (e.g., IP-10 and MCP-5) measured in
the brain at 24-hr post-dose, and these cytokine levels were
restored to the baseline level at 96-hr post-dose. ATV:TREM2 #3 at
3 mg/kg had maximum effect on brain cytokine levels (FIGS. 18A and
18B).
[0418] ATV:TREM12 Increases Brain CSF1R Level in TB36/hTfR KI
Mice
[0419] To determine the effect of ATV:TREM2 on the level of glial
marker CSF1R, CSF1R protein level was measured in brain lysates
(prepared by Cell Signaling lysis buffer #9803) of TB36/hTfR KI
mice using a commercial ELISA kit (Abcam ab240681). We found that
increasing doses of ATV:TREM2 #3 resulted in an increased CSF1R
protein level at 1 day after treatment and 4 days after treatment
(FIG. 19).
[0420] Plasma PK Profile of ATV:TREM2
[0421] Proportional increases in plasma PK were observed with doses
of ATV:TREM2 #3 ranging from 1 mg/kg to 100 mg/kg. At 24 hours, the
plasma concentrations of matched doses of 30 mg/kg of ATV:TREM2 #3,
reference antibody #2, and TREM2 Ab were not significantly
different. Reference antibody #2 appeared to have a lower clearance
compared to ATV:TREM2 #3 and TREM2 Ab at the same dose (FIG.
20).
[0422] Brain PK Profile of ATV:TREM2
[0423] At 24 hours, the brain concentration of 10 mg/kg of
ATV:TREM2 #3 was similar to TREM2 Ab at 30 mg/kg, and the brain
concentration of 1-3 mg/kg of ATV:TREM2 #3 was similar to reference
antibody #2 at 30 mg/kg. More than proportional increases were
observed at doses of 3 mg/kg to 10 mg/kg of ATV:TREM2 #3, and
proportional increases were observed at other doses. At 96 hours,
the brain concentration of 10 mg/kg of ATV:TREM2 #3 was similar to
TREM2 Ab and the reference antibody #2 at 30 mg/kg (FIG. 21).
Overall, brain uptake of ATV:TREM2 #3 is more efficient than TREM2
Ab and reference antibody #2.
TABLE-US-00008 TABLE 8 Informal Sequence Listing SEQ ID NO Sequence
Description 1 MEPLRLLILLFVTELSGAHNTTVFQGVAGQSLQVSCPYDSMKH Human
TREM2 protein WGRRKAWCRQLGEKGPCQRVVSTHNLWLLSFLRRWNGSTAIT
DDTLGGTLTITLRNLQPHDAGLYQCQSLHGSEADTLRKVLVEVL
ADPLDHRDAGDLWFPGESESFEDAHVEHSISRSLLEGEIPFPPTSI
LLLLACIFLIKILAASALWAAAWHGQKPGTHPPSELDCGHDPGY QLQTLPGLRDT 2
EVKLLDSGGGLVQAGGSLRLSCAGSGFTFTDFYMSWIRQPPGKA CL0020306 V.sub.H
PEWLGVIRNKANGYTAGYNPSVKGRFTISRDNTQNILYLQMNTL
RAEDTAIYYCARLSYGFDYWGQGVMVTVSS 3
DIVMTQGALPNPVPSGESASITCQSSKSLLHSNGKTYLNWYLQR CL0020306 V.sub.L
PGQSPQLLIYWMSTRASGVSDRFSGSGSGTDFTLKISSVEAEDVG
VYYCQQFLEFPFTFGSGTKLEIK 4 GFTFTDFYMS CL0020306 CDR-H1; CL0020164
CDR-H1; CDR-H1 for CL0020188 and variants CL0020188-1, CL0020188-2,
CL0020188-3, CL0020188-4, CL0020188-5, CL0020188-6, CL0020188-7,
and CL0020188-8 5 VIRNKANGYTAGYNPSVKG CL0020306 CDR-H2; CDR-H2 for
CL0020188 and variants CL0020188-1, CL0020188-2, CL0020188-3, and
CL0020188-4 6 ARLSYGFDY CL0020306 CDR-H3 7 QSSKSLLHSNGKTYLN
CL0020306 CDR-L1; CL0020164 CDR-L1; CL0020307 CDR-L1; CL0020307-1
CDR-L1; CDR-L1 for CL0020188 and variants CL0020188-1, CL0020188-2,
CL0020188-5, and CL0020188-6 8 WMSTRAS CL0020306 CDR-L2; CL0020307
CDR-L2; CL0020307-1 CDR-L2; CL0020164 CDR-L2; CDR- L2 for CL0020188
and variants CL0020188-1, CL0020188-2, CL0020188-3, CL0020188-4,
CL0020188-5, CL0020188-6, CL0020188-7, and CL0020188-8 9 QQFLEFPFT
CL0020306 CDR-L3; CL0020307 CDR-L3; CL0020307-1 CDR-L3 10
EVKLLESGGGLVQPGGSLRLSCAASGFTFTNFYMSWIRQPPGRA CL0020307 V.sub.H
PEWLGVIRNRPNGYTTDYNPSVKGRFTISRDNTQNILYLQMSTL
RADDTAFYYCTRLTYGFDYWGQGVMVTVSS 11
DIVMTQGALPNPVPSGESASITCQSSKSLLHSNGKTYLNWYLQR CL0020307 V.sub.L
PGQSPQLLIYWMSTRASGVSDRFSGSGSGTDFTLKISSVEAEVVG
VYYCQQFLEFPFTFGSGTKLEIK 12 GFTFTNFYMS CL0020307 CDR-H1 13
VIRNRPNGYTTDYNPSVKG CL0020307 CDR-H2 14 TRLTYGFDY CL0020307 CDR-H3
15 EVKLLDSGGGLVQAGGSLRLSCAGSGFTFTDFYMSWIRQPPGKA CL0020188 V.sub.H
PEWLGVIRNKANGYTAGYNPSVKGRFTISRDNTQNILYLQMNTL
RAEDTAIYYCARLTYGFDYWGQGVMVTVSS 16
DIVMTQGALPNPVPSGESASITCQSSKSLLHSNGKTYLNWYLQR CL0020188 V.sub.L
PGQSPQLLIYWMSTRASGVSDRFSGSGSGTDFTLKISSVEAEDVG
VYYCQQFLEYPFTFGSGTKLEIK 17 ARLTYGFDY CDR-H3 for CL0020188 and
variants CL0020188-1, CL0020188-2, CL0020188-3, CL0020188-4,
CL0020188-5, CL0020188-6, CL0020188-7, and CL0020188-8; CL0020164
CDR-H3 18 QQFLEYPFT CDR-L3 for CL0020188 and variants CL0020188-1,
CL0020188-2, CL0020188-3, CL0020188-4, CL0020188-5, CL0020188-6,
CL0020188-7, and CL0020188-8 19
EVQLVESGGGLVQPGGSLRLSCAASGFTFTDFYMSWVRQAPGK CL0020188-1 V.sub.H;
GLEWVSVIRNKANGYTAGYNPSVKGRFTISRDNSKNTLYLQMN CL0020188-3 V.sub.H
SLRAEDTAVYYCARLTYGFDYWGQGTLVTVSS 20
DIVMTQTPLSLPVTPGEPASISCQSSKSLLHSNGKTYLNWYLQKP CL0020188-1 V.sub.L;
GQSPQLLIYWMSTRASGVPDRFSGSGSGTDFTLKISRVEAEDVG CL0020188-2 V.sub.L;
VYYCQQFLEYPFTFGQGTKVEIK CL0020188-5 V.sub.L; CL0020188-6 V.sub.L 21
EVQLVESGGGLVQPGGSLRLSCAGSGFTFTDFYMSWVRQAPGK CL0020188-2 V.sub.H;
GLEWVSVIRNKANGYTAGYNPSVKGRFTISRDNSKNTLYLQMN CL0020188-4 V.sub.H
SLRAEDTAVYYCARLTYGFDYWGQGTLVTVSS 22
DIVMTQTPLSLPVTPGEPASISCQSSKSLLHSTGKTYLNWYLQKP CL0020188-3 V.sub.L;
GQSPQLLIYWMSTRASGVPDRFSGSGSGTDFTLKISRVEAEDVG CL0020188-4 V.sub.L;
VYYCQQFLEYPFTFGQGTKVEIK CL0020188-7 V.sub.L; CL0020188-8 V.sub.L 23
QSSKSLLHSTGKTYLN CDR-L1 for variants CL0020188-3, CL0020188-4,
CL0020188-7, and CL0020188-8 24
EVQLVESGGGLVQPGGSLRLSCAASGFTFTDFYMSWVRQAPGK CL0020188-5 V.sub.H;
GLEWVSVIRNKANAYTAGYNPSVKGRFTISRDNSKNTLYLQMN CL0020188-7 V.sub.H
SLRAEDTAVYYCARLTYGFDYWGQGTLVTVSS 25 VIRNKANAYTAGYNPSVKG CDR-H2 for
variants CL0020188-5, CL0020188-6, CL0020188-7, and CL0020188-8 26
EVQLVESGGGLVQPGGSLRLSCAGSGFTFTDFYMSWVRQAPGK CL0020188-6 V.sub.H;
GPEWLSVIRNKANAYTAGYNPSVKGRFTISRDNSKNTLYLQMN CL0020188-8 V.sub.H
SLRAEDTAVYYCARLTYGFDYWGQGTLVTVSS 27
DIVMTQSPDSLAVSLGERATINCQSSKSLLHSNGKTYLNWYQQK CL0020307-1 V.sub.L
PGQPPKLLIYWMSTRASGVPDRFSGSGSGTDFTLTISSLQAEDVA
VYYCQQFLEFPFTFGQGTKVEIK 28 G-F-T-F-T-.alpha..sub.6-F-Y-M-S, wherein
.alpha..sub.6 is D or N CDR-H1 consensus sequence 29
V-I-R-N-.beta..sub.5-.beta..sub.6-N-.beta..sub.8-Y-T-.beta..sub.11-.bet-
a..sub.12-Y-N-P-S-V-K-G, CDR-H2 consensus sequence wherein
.beta..sub.5 is K or R; .beta..sub.6 is A or P; .beta..sub.8 is G
or A; .beta..sub.11 is A or T; and .beta..sub.12 is G or D 30
.gamma..sub.1-R-L-.gamma..sub.4-Y-G-F-D-Y, wherein .gamma..sub.1 is
A or T; CDR-H3 consensus sequence and .gamma..sub.4 is T or S 31
Q-S-S-K-S-L-L-H-S-.delta..sub.10-G-K-T-Y-L-N, CDR-L1 consensus
sequence wherein .delta..sub.10 is N or T 32
Q-Q-F-L-E-.PHI..sub.6-P-F-T, wherein .PHI..sub.6 is Y or F CDR-L3
consensus sequence 33 000 34 GGGGS Linker sequence 35 HHHHHH 6X-His
tag 36 ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSG Heavy chain
constant domain ALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKP 1
(CH1) SNTKVDKKVEPKSCDKTHTCPPCP 37
RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVD light chain constant
domain NALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACE (CL)
VTHQGLSSPVTKSFNRGEC 38
APELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKF Wild-type human Fc
sequence NWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNG positions
231-447 EU index KEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKN
numbering QVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFF
LYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK 39
APEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKF Fc polypeptide with
hole, NWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNG LALA, and LS
mutations KEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKN
QVSLSCAVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFF
LVSKLTVDKSRWQQGNVFSCSVLHEALHSHYTQKSLSLSPGK 40
APELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKF Clone 35.21
NWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNG
KEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKN
QVSLTCLVKGFYPSDIAVWWESYG1EWSSYKTTPPVLDSDGSFF
LYSKLTVTKEEWQQGFVFSCSVMHEALHNHYTQKSLSLSPGK 41
APEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKF Clone CH3C.35.21 Fc
NWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNG polypeptide with knob,
LALA, KEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKN and LS
mutations QVSLWCLVKGFYPSDIAVWWESYGTEWSSYKTTPPVLDSDGSFF
LYSKLTVTKEEWQQGFVFSCSVLHEALHSHYTQKSLSLSPGK 42
EVQLVESGGGLVQPGGSLRLSCAASGFTFTDFYMSWVRQAPGK anti-TREM2/CH35.21
heavy GLEWVSVIRNKANAYTAGYNPSVKGRFTISRDNSKNTLYLQMN chain with knob,
LALA and SLRAEDTAVYYCARLTYGFDYWGQGTLVTVSSASTKGPSVFPL LS mutations
APSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPA
VLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEP
KSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCV
VVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVS
VLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQ
VYTLPPSRDELTKNQVSLWCLVKGFYPSDIAVWWESYGTEWSS
YKTTPPVLDSDGSFFLYSKLTVTKEEWQQGFVFSCSVLHEALHS HYTQKSLSLSPGK 43
APELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKF Clone CH3C.35.23.1.1
NWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNG
KEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKN
QVSLTCLVKGFYPSDIAVEWESFGTEWSNYKTTPPVLDSDGSFF
LYSKLTVSKEEWQQGFVFSCSVMHEALHNHYTQKSLSLSPGK 44
APEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKF Clone CH3C.35.23.1.1
Fc NWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNG polypeptide with knob,
LALA, KEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKN and LS
mutations QVSLWCLVKGFYPSDIAVEWESFGIEWSNYKTTPPVLDSDGSFF
LYSKLTVSKEEWQQGFVFSCSVLHEALHSHYTQKSLSLSPGK 45
EVQLVESGGGLVQPGGSLRLSCAASGFTFTDFYMSWVRQAPGK anti-TREM2/CH35.23.1.1
GLEWVSVIRNKANAYTAGYNPSVKGRFTISRDNSKNTLYLQMN heavy chain with knob,
LALA SLRAEDTAVYYCARLTYGFDYWGQGTLVTVSSASTKGPSVFPL and LS mutations
APSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPA
VLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEP
KSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCV
VVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVS
VLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQ
VYTLPPSRDELTKNQVSLWCLVKGFYPSDIAVEWESFGTEWSNY
KTTPPVLDSDGSFFLYSKLTVSKEEWQQGFVFSCSVLHEALHSH YTQKSLSLSPGK 46
APELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKF Clone CH3C.35.23.3
NWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNG
KEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKN
QVSLTCLVKGFYPSDIAVEWESYGIEWVNYKTTPPVLDSDGSFF
LYSKLTVTKEEWQQGFVFSCSVMHEALHNHYTQKSLSLSPGK 47
APEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKF CH3C.35.23.3 Fc
polypeptide NWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNG with knob,
LALA, and LS KEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKN
mutations QVSLWCLVKGFYPSDIAVEWESYGTEWVNYKTTPPVLDSDGSFF
LYSKLTVTKEEWQQGFVFSCSVLHEALHSHYTQKSLSLSPGK
48 EVQLVESGGGLVQPGGSLRLSCAASGFTFTDFYMSWVRQAPGK anti-TREM2/CH35.23.3
GLEWVSVIRNKANAYTAGYNPSVKGRFTISRDNSKNTLYLQMN heavy chain with knob,
LALA SLRAEDTAVYYCARLTYGFDYWGQGTLVTVSSASTKGPSVFPL and LS mutations
APSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPA
VLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEP
KSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCV
VVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVS
VLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQ
VYTLPPSRDELTKNQVSLWCLVKGFYPSDIAVEWESYGTEWVN
YKTTPPVLDSDGSFFLYSKLTVTKEEWQQGFVFSCSVLHEALHS HYTQKSLSLSPGK 49
APELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKF Clone CH3C.35.23.4
NWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNG
KEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKN
QVSLTCLVKGFYPSDIAVEWESYGTEWSNYKTTPPVLDSDGSFF
LYSKLTVSKEEWQQGFVFSCSVMHEALHNHYTQKSLSLSPGK 50
APEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKF CH3C.35.23.4 Fc
polypeptide NWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNG with knob,
LALA, and LS KEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKN
mutations QVSLWCLVKGFYPSDIAVEWESYGTEWSNYKTTPPVLDSDGSFF
LYSKLTVSKEEWQQGFVFSCSVLHEALHSHYTQKSLSLSPGK 51
EVQLVESGGGLVQPGGSLRLSCAASGFTFTDFYMSWVRQAPGK anti-TREM2/CH35.23.4
GLEWVSVIRNKANAYTAGYNPSVKGRFTISRDNSKNTLYLQMN heavy chain with knob,
LALA SLRAEDTAVYYCARLTYGFDYWGQGTLVTVSSASTKGPSVFPL and LS mutations
APSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPA
VLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEP
KSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCV
VVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVS
VLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQ
VYTLPPSRDELTKNQVSLWCLVKGFYPSDIAVEWESYGTEWSN
YKTTPPVLDSDGSFFLYSKLTVSKEEWQQGFVFSCSVLHEALHS HYTQKSLSLSPGK 52
EVQLVESGGGLVQPGGSLRLSCAASGFTFTDFYMSWVRQAPGK anti-TREM2 heavy chain
with GLEWVSVIRNKANAYTAGYNPSVKGRFTISRDNSKNTLYLQMN hole and LS
mutations SLRAEDTAVYYCARLTYGFDYWGQGTLVTVSSASTKGPSVFPL
APSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPA
VLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEP
KSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCV
VVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVS
VLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQ
VYTLPPSRDELTKNQVSLSCAVKGFYPSDIAVEWESNGQPENN
YKTTPPVLDSDGSFFLVSKLTVDKSRWQQGNVFSCSVLHEALHS HYTQKSLSLSPGK 53
EVQLVESGGGLVQPGGSLRLSCAASGFTFTDFYMSWVRQAPGK anti-TREM2 heavy chain
with GLEWVSVIRNKANAYTAGYNPSVKGRFTISRDNSKNTLYLQMN hole, LALA and LS
mutations SLRAEDTAVYYCARLTYGFDYWGQGTLVTVSSASTKGPSVFPL
APSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPA
VLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEP
KSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCV
VVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVS
VLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQ
VYTLPPSRDELTKNQVSLSCAVKGFYPSDIAVEWESNGQPENN
YKTTPPVLDSDGSFFLVSKLTVDKSRWQQGNVFSCSVLHEALHS HYTQKSLSLSPGK 54
DIVMTQTPLSLPVTPGEPASISCQSSKSLLHSTGKTYLNWYLQKP anti-TREM2 light
chain GQSPQLLIYWMSTRASGVPDRFSGSGSGTDFTLKISRVEAEDVG
VYYCQQFLEYPFTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGT
ASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDST
YSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC 55
NSVIIVDKNGRLVYLVENPGGYVAYSKAATVTGKLVHANFGTK Human TfR apical domain
KDFEDLYTPVNGSIVIVRAGKITFAEKVANAESLNAIGVLIYMDQ
TKFPIVNAELSFFGHAHLGTGDPYTPGFPSFNHTQFPPSRSSGLPN
IPVQTISRAAAEKLFGNMEGDCPSDWKTDSTCRMVTSESKNVKL TVS 56 GGGGSGGGGS
Linker 57 DKTHTCPPCP Portion of human IgG1 hinge sequence 58
YxTEWSS CH3 motif (TfR-binding) 59 TxxExxxxF CH3 motif
(TfR-binding) 60 MMDQARSAFSNLFGGEPLSYTRFSLARQVDGDNSHVEMKLGV Cyno
TfR DEEENTDNNTKANGTKPKRCGGNICYGTIAVIIFFLIGFMIGYLG
YCKGVEPKTECERLAGTESPAREEPEEDFPAAPRLYWDDLKRKL
SEKLDTTDFTSTIKLLNENLYVPREAGSQKDENLALYIENQFREF
KLSKVWRDQHFVKIQVKDSAQNSVIIVDKNGGLVYLVENPGGY
VAYSKAATVTGKLVHANFGTKKDFEDLDSPVNGSIVIVRAGKIT
FAEKVANAESLNAIGVLIYMDQTKFPIVKADLSFFGHAHLGTGD
PYTPGFPSFNHTQFPPSQSSGLPNIPVQTISRAAAEKLFGNMEGDC
PSDWKTDSTCKMVTSENKSVKLTVSNVLKETKILNIFGVIKGFV
EPDHYVVVGAQRDAWGPGAAKSSVGTALLLKLAQMFSDMVLK
DGFQPSRSIIFASWSAGDFGSVGATEWLEGYLSSLHLKAFTYINL
DKAVLGTSNFKVSASPLLYTLIEKTMQDVKHPVTGRSLYQDSN
WASKVEKLTLDNAAFPFLAYSGIPAVSFCFCEDTDYPYLGTTMD
TYKELVERIPELNKVARAAAEVAGQFVIKLTHDTELNLDYERYN
SQLLLFLRDLNQYRADVKEMGLSLQWLYSARGDFFRATSRLTT
DFRNAEKRDKFVMKKLNDRVMRVEYYFLSPYVSPKESPFRHVF
WGSGSHTLSALLESLKLRRQNNSAFNETLFRNQLALATWTIQGA ANALSGDVWDIDNEF 61
APELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKF Fc polypeptide with
hole and NWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNG LS mutations
KEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKN
QVSLSCAVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFF
LVSKLTVDKSRWQQGNVFSCSVLHEALHSHYTQKSLSLSPGK 62
MMDQARSAFSNLFGGEPLSYTRFSLARQVDGDNSHVEMKLAV Human transferrin
receptor DEEENADNNTKANVTKPKRCSGSICYGTIAVIVFFLIGFMIGYLG protein 1
(TFR1) YCKGVEPKTECERLAGTESPVREEPGEDFPAARRLYWDDLKRK
LSEKLDSTDFTGTIKLLNENSYVPREAGSQKDENLALYVENQFR
EFKLSKVWRDQHFVKIQVKDSAQNSVIIVDKNGRLVYLVENPG
GYVAYSKAATVTGKLVHANFGTKKDFEDLYTPVNGSIVIVRAG
KITFAEKVANAESLNAIGVLIYMDQTKFPIVNAELSFFGHAHLGT
GDPYTPGFPSFNHTQFPPSRSSGLPNIPVQTISRAAAEKLFGNMEG
DCPSDWKTDSTCRMVTSESKNVKLTVSNVLKEIKILNIFGVIKGF
VEPDHYVVVGAQRDAWGPGAAKSGVGTALLLKLAQMFSDMV
LKDGFQPSRSIIFASWSAGDFGSVGATEWLEGYLSSLHLKAFTYI
NLDKAVLGTSNFKVSASPLLYTLIEKTMQNVKHPVTGQFLYQDS
NVVASKVEKLTLDNAAFPFLAYSGIPAVSFCFCEDTDYPYLGTTM
DTYKELIERIPELNKVARAAAEVAGQFVIKLTHDVELNLDYERY
NSQLLSFVRDLNQYRADIKEMGLSLQWLYSARGDFFRATSRLTT
DFGNAEKTDRFVMKKLNDRVMRVEYHFLSPYVSPKESPFRHVF
WGSGSHTLPALLENLKLRKQNNGAFNETLFRNQLALATWTIQG AANALSGDVWDIDNEF 63
APEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKF Fc polypeptide with
hole, NWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNG LALA, and LS
mutations, KEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKN truncated
QVSLSCAVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFF
LVSKLTVDKSRWQQGNVFSCSVLHEALHSHYTQKSLSLSPG 64
APEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKF Clone CH3C.35.21 Fc
NWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNG polypeptide with knob,
LALA, KEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKN and LS
mutations, truncated QVSLWCLVKGFYPSDIAVWWESYGTEWSSYKTTPPVLDSDGSFF
LYSKLTVTKEEWQQGFVFSCSVLHEALHSHYTQKSLSLSPG 65
EVQLVESGGGLVQPGGSLRLSCAASGFTFTDFYMSWVRQAPGK anti-TREM2/CH35.21
heavy GLEWVSVIRNKANAYTAGYNPSVKGRFTISRDNSKNTLYLQMN chain with knob,
LALA and SLRAEDTAVYYCARLTYGFDYWGQGTLVTVSSASTKGPSVFPL LS mutations,
truncated APSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPA
VLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEP
KSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCV
VVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVS
VLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQ
VYTLPPSRDELTKNQVSLWCLVKGFYPSDIAVWWESYGTEWSS
YKTTPPVLDSDGSFFLYSKLTVTKEEWQQGFVFSCSVLHEALHS HYTQKSLSLSPG 66
APEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKF Clone CH3C.35.23.1.1
Fc NWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNG polypeptide with knob,
LALA, KEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKN and LS
mutations, truncated QVSLWCLVKGFYPSDIAVEWESFGIEWSNYKTTPPVLDSDGSFF
LYSKLTVSKEEWQQGFVFSCSVLHEALHSHYTQKSLSLSPG 67
EVQLVESGGGLVQPGGSLRLSCAASGFTFTDFYMSWVRQAPGK anti-TREM2/CH35.23.1.1
GLEWVSVIRNKANAYTAGYNPSVKGRFTISRDNSKNTLYLQMN heavy chain with knob,
LALA SLRAEDTAVYYCARLTYGFDYWGQGTLVTVSSASTKGPSVFPL and LS mutations,
truncated APSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPA
VLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEP
KSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCV
VVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVS
VLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQ
VYTLPPSRDELTKNQVSLWCLVKGFYPSDIAVEWESFGTEWSNY
KTTPPVLDSDGSFFLYSKLTVSKEEWQQGFVFSCSVLHEALHSH YTQKSLSLSPG 68
APEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKF CH3C.35.23.3 Fc
polypeptide NWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNG with knob,
LALA, and LS KEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKN
mutations, truncated QVSLWCLVKGFYPSDIAVEWESYGTEWVNYKTTPPVLDSDGSFF
LYSKLTVTKEEWQQGFVFSCSVLHEALHSHYTQKSLSLSPG 69
EVQLVESGGGLVQPGGSLRLSCAASGFTFTDFYMSWVRQAPGK anti-TREM2/CH35.23.3
GLEWVSVIRNKANAYTAGYNPSVKGRFTISRDNSKNTLYLQMN heavy chain with knob,
LALA SLRAEDTAVYYCARLTYGFDYWGQGTLVTVSSASTKGPSVFPL and LS mutations,
truncated APSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPA
VLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEP
KSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCV
VVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVS
VLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQ
VYTLPPSRDELTKNQVSLWCLVKGFYPSDIAVEWESYGTEWVN
YKTTPPVLDSDGSFFLYSKLTVTKEEWQQGFVFSCSVLHEALHS HYTQKSLSLSPG 70
APEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKF CH3C.35.23.4 Fc
polypeptide NWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNG with knob,
LALA, and LS KEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKN
mutations, truncated QVSLWCLVKGFYPSDIAVEWESYGTEWSNYKTTPPVLDSDGSFF
LYSKLTVSKEEWQQGFVFSCSVLHEALHSHYTQKSLSLSPG 71
EVQLVESGGGLVQPGGSLRLSCAASGFTFTDFYMSWVRQAPGK anti-TREM2/CH35.23.4
GLEWVSVIRNKANAYTAGYNPSVKGRFTISRDNSKNTLYLQMN heavy chain with knob,
LALA SLRAEDTAVYYCARLTYGFDYWGQGTLVTVSSASTKGPSVFPL and LS mutations,
truncated APSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPA
VLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEP
KSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCV
VVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVS
VLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQ
VYTLPPSRDELTKNQVSLWCLVKGFYPSDIAVEWESYGTEWSN
YKTTPPVLDSDGSFFLYSKLTVSKEEWQQGFVFSCSVLHEALHS HYTQKSLSLSPG 72
EVQLVESGGGLVQPGGSLRLSCAASGFTFTDFYMSWVRQAPGK anti-TREM2 heavy chain
with GLEWVSVIRNKANAYTAGYNPSVKGRFTISRDNSKNTLYLQMN hole and LS
mutations, SLRAEDTAVYYCARLTYGFDYWGQGTLVTVSSASTKGPSVFPL truncated
APSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPA
VLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEP
KSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCV
VVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVS
VLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQ
VYTLPPSRDELTKNQVSLSCAVKGFYPSDIAVEWESNGQPENN
YKTTPPVLDSDGSFFLVSKLTVDKSRWQQGNVFSCSVLHEALHS HYTQKSLSLSPG 73
EVQLVESGGGLVQPGGSLRLSCAASGFTFTDFYMSWVRQAPGK anti-TREM2 heavy chain
with GLEWVSVIRNKANAYTAGYNPSVKGRFTISRDNSKNTLYLQMN hole, LALA and LS
mutations, SLRAEDTAVYYCARLTYGFDYWGQGTLVTVSSASTKGPSVFPL truncated
APSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPA
VLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEP
KSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCV
VVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVS
VLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQ
VYTLPPSRDELTKNQVSLSCAVKGFYPSDIAVEWESNGQPENN
YKTTPPVLDSDGSFFLVSKLTVDKSRWQQGNVFSCSVLHEALHS HYTQKSLSLSPG 74
QVQLVQSGAEVKKPGASVKVSCKASGYAFSSQWMNWVRQAP Reference antibody #1
heavy GQRLEWIGRIYPGGGDTNYAGKFQGRVTITADTSASTAYMELSS chain
LRSEDTAVYYCARLLRNQPGESYAMDYWGQGTLVTVSSASTK
GPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTS
GVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTK
VDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISR
TPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYN
STYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAK
GQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESN
GQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSV MHEALHNHYTQKSLSLSPGK 75
DVVMTQSPDSLAVSLGERATINCRSSQSLVHSNRYTYLHWYQQ Reference antibody #1
and
#2 KPGQSPKLLIYKVSNRFSGVPDRFSGSGSGTDFTLKISRVEAEDV light chain
GVYYCSQSTRVPYTFGQGTKLEIKRTVAAPSVFIFPPSDEQLKSG
TASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDS
TYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC 76
QVQLVQSGAEVKKPGASVKVSCKASGYAFSSQWMNWVRQAP Reference antibody #2
heavy GQRLEWIGRIYPGGGDTNYAGKFQGRVTITADTSASTAYMELSS chain
LRSEDTAVYYCARLLRNQPGESYAMDYWGQGTLVTVSSASTK
GPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTS
GVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTK
VDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISR
TPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYN
STYRVVSVLTVLHQDWLNGKEYKCKVSNKALPASIEKTISKAK
GQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESN
GQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSV MHGALHNHYTQKSLSLSPGK 77
QVQLQESGPGLVKPSETLSLTCAVSGYSITSDYAWNWIRQPPGK Isotype control for
reference GLEWIGYMSYSGSTRYNPSLRSRVTISVDTSKNQFSLKLSSVTAA antibodies
#1 and #2, heavy DTAVYYCARGWPLAYWGQGTLVTVSSASTKGPSVFPLAPSSKS chain
TSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSG
LYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDK
THTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVS
HEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVL
HQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPP
SRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPV
LDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKS LSLSPGK 78
EIVMTQSPATLSLSPGERATLSCSASSSVSYMYWYQQKPGQAPR Isotype control for
reference LLIYDTSNLASGIPARFSGSGSGTDFTLTISSLQPEDFAVYYCQQW antibodies
#1 and #2, light SSYPPITFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLL
chain NNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTL
TLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC 79
QVTLRESGPALVKPTQTLTLTCTFSGFSLSTSGMSVGWIRQPPGK Isotype control for
ALEWLADIWWDDKKDYNPSLKSRLTISKDTSKNQVVLKVTNM ATV: TREM2 #1, heavy
chain EPADTATYYCARSMITNWYFDVWGAGTTVTVSSASTKGPSVFP with knob, LALA
LAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFP
AVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKV
EPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVT
CVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRV
VSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREP
QVYTLPPSRDELTKNQVSLWCLVKGFYPSDIAVWWESYGTEWS
SYKTTPPVLDSDGSFFLYSKLTVTKEEWQQGFVFSCSVMHEALH NHYTQKSLSLSPGK 80
QVTLRESGPALVKPTQTLTLTCTFSGFSLSTSGMSVGWIRQPPGK Isotype control for
ALEWLADIWWDDKKDYNPSLKSRLTISKDTSKNQVVLKVTNM ATV: TREM2 #3, heavy
chain EPADTATYYCARSMITNWYFDVWGAGTTVTVSSASTKGPSVFP with knob, LALA
LAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFP
AVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKV
EPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVT
CVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRV
VSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREP
QVYTLPPSRDELTKNQVSLWCLVKGFYPSDIAVEWESYGTEWV
NYKTTPPVLDSDGSFFLYSKLTVTKEEWQQGFVFSCSVMHEALH NHYTQKSLSLSPGK 81
QVTLRESGPALVKPTQTLTLTCTFSGFSLSTSGMSVGWIRQPPGK Isotype control for
ALEWLADIWWDDKKDYNPSLKSRLTISKDTSKNQVVLKVTNM ATV: TREM #1 and #3,
heavy EPADTATYYCARSMITNWYFDVWGAGTTVTVSSASTKGPSVFP chain with hole,
LALA LAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFP
AVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKV
EPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVT
CVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRV
VSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREP
QVYTLPPSRDELTKNQVSLSCAVKGFYPSDIAVEWESNGQPENN
YKTTPPVLDSDGSFFLVSKLTVDKSRWQQGNVFSCSVMHEALH NHYTQKSLSLSPGK 82
DIQMTQSPSTLSASVGDRVTITCKCQLSVGYMHWYQQKPGKAP Isotype control for
KLLIYDTSKLASGVPSRFSGSGSGTEFTLTISSLQPDDFATYYCFQ ATV: TREM2 #1 and
#3, light GSGYPFTFGGGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLL chain
NNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTL
TLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC 83
EVQLVESGGGLVQPGGSLRLSCAASGFTFTDFYMSWVRQAPGK anti-TREM2 heavy chain
with GLEWVSVIRNKANAYTAGYNPSVKGRFTISRDNSKNTLYLQMN LALA mutations
SLRAEDTAVYYCARLTYGFDYWGQGTLVTVSSASTKGPSVFPL
APSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPA
VLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEP
KSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCV
VVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVS
VLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQ
VYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNY
KTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHN HYTQKSLSLSPGK 84
APELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKF Fc polypeptide with
hole and NWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNG LS mutations,
truncated KEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKN
QVSLSCAVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFF
LVSKLTVDKSRWQQGNVFSCSVLHEALHSHYTQKSLSLSPG
Sequence CWU 1 SEQUENCE LISTING <160> NUMBER OF SEQ ID
NOS: 84 <210> SEQ ID NO 1 <211> LENGTH: 230 <212>
TYPE: PRT <213> ORGANISM: Homo sapiens <400> SEQUENCE:
1 Met Glu Pro Leu Arg Leu Leu Ile Leu Leu Phe Val Thr Glu Leu Ser 1
5 10 15 Gly Ala His Asn Thr Thr Val Phe Gln Gly Val Ala Gly Gln Ser
Leu 20 25 30 Gln Val Ser Cys Pro Tyr Asp Ser Met Lys His Trp Gly
Arg Arg Lys 35 40 45 Ala Trp Cys Arg Gln Leu Gly Glu Lys Gly Pro
Cys Gln Arg Val Val 50 55 60 Ser Thr His Asn Leu Trp Leu Leu Ser
Phe Leu Arg Arg Trp Asn Gly 65 70 75 80 Ser Thr Ala Ile Thr Asp Asp
Thr Leu Gly Gly Thr Leu Thr Ile Thr 85 90 95 Leu Arg Asn Leu Gln
Pro His Asp Ala Gly Leu Tyr Gln Cys Gln Ser 100 105 110 Leu His Gly
Ser Glu Ala Asp Thr Leu Arg Lys Val Leu Val Glu Val 115 120 125 Leu
Ala Asp Pro Leu Asp His Arg Asp Ala Gly Asp Leu Trp Phe Pro 130 135
140 Gly Glu Ser Glu Ser Phe Glu Asp Ala His Val Glu His Ser Ile Ser
145 150 155 160 Arg Ser Leu Leu Glu Gly Glu Ile Pro Phe Pro Pro Thr
Ser Ile Leu 165 170 175 Leu Leu Leu Ala Cys Ile Phe Leu Ile Lys Ile
Leu Ala Ala Ser Ala 180 185 190 Leu Trp Ala Ala Ala Trp His Gly Gln
Lys Pro Gly Thr His Pro Pro 195 200 205 Ser Glu Leu Asp Cys Gly His
Asp Pro Gly Tyr Gln Leu Gln Thr Leu 210 215 220 Pro Gly Leu Arg Asp
Thr 225 230 <210> SEQ ID NO 2 <211> LENGTH: 118
<212> TYPE: PRT <213> ORGANISM: Artificial Sequence
<220> FEATURE: <221> NAME/KEY: source <223> OTHER
INFORMATION: /note="Description of Artificial Sequence: Synthetic
polypeptide" <400> SEQUENCE: 2 Glu Val Lys Leu Leu Asp Ser
Gly Gly Gly Leu Val Gln Ala Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser
Cys Ala Gly Ser Gly Phe Thr Phe Thr Asp Phe 20 25 30 Tyr Met Ser
Trp Ile Arg Gln Pro Pro Gly Lys Ala Pro Glu Trp Leu 35 40 45 Gly
Val Ile Arg Asn Lys Ala Asn Gly Tyr Thr Ala Gly Tyr Asn Pro 50 55
60 Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Thr Gln Asn Ile
65 70 75 80 Leu Tyr Leu Gln Met Asn Thr Leu Arg Ala Glu Asp Thr Ala
Ile Tyr 85 90 95 Tyr Cys Ala Arg Leu Ser Tyr Gly Phe Asp Tyr Trp
Gly Gln Gly Val 100 105 110 Met Val Thr Val Ser Ser 115 <210>
SEQ ID NO 3 <211> LENGTH: 112 <212> TYPE: PRT
<213> ORGANISM: Artificial Sequence <220> FEATURE:
<221> NAME/KEY: source <223> OTHER INFORMATION:
/note="Description of Artificial Sequence: Synthetic polypeptide"
<400> SEQUENCE: 3 Asp Ile Val Met Thr Gln Gly Ala Leu Pro Asn
Pro Val Pro Ser Gly 1 5 10 15 Glu Ser Ala Ser Ile Thr Cys Gln Ser
Ser Lys Ser Leu Leu His Ser 20 25 30 Asn Gly Lys Thr Tyr Leu Asn
Trp Tyr Leu Gln Arg Pro Gly Gln Ser 35 40 45 Pro Gln Leu Leu Ile
Tyr Trp Met Ser Thr Arg Ala Ser Gly Val Ser 50 55 60 Asp Arg Phe
Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Lys Ile 65 70 75 80 Ser
Ser Val Glu Ala Glu Asp Val Gly Val Tyr Tyr Cys Gln Gln Phe 85 90
95 Leu Glu Phe Pro Phe Thr Phe Gly Ser Gly Thr Lys Leu Glu Ile Lys
100 105 110 <210> SEQ ID NO 4 <211> LENGTH: 10
<212> TYPE: PRT <213> ORGANISM: Artificial Sequence
<220> FEATURE: <221> NAME/KEY: source <223> OTHER
INFORMATION: /note="Description of Artificial Sequence: Synthetic
peptide" <400> SEQUENCE: 4 Gly Phe Thr Phe Thr Asp Phe Tyr
Met Ser 1 5 10 <210> SEQ ID NO 5 <211> LENGTH: 19
<212> TYPE: PRT <213> ORGANISM: Artificial Sequence
<220> FEATURE: <221> NAME/KEY: source <223> OTHER
INFORMATION: /note="Description of Artificial Sequence: Synthetic
peptide" <400> SEQUENCE: 5 Val Ile Arg Asn Lys Ala Asn Gly
Tyr Thr Ala Gly Tyr Asn Pro Ser 1 5 10 15 Val Lys Gly <210>
SEQ ID NO 6 <211> LENGTH: 9 <212> TYPE: PRT <213>
ORGANISM: Artificial Sequence <220> FEATURE: <221>
NAME/KEY: source <223> OTHER INFORMATION: /note="Description
of Artificial Sequence: Synthetic peptide" <400> SEQUENCE: 6
Ala Arg Leu Ser Tyr Gly Phe Asp Tyr 1 5 <210> SEQ ID NO 7
<211> LENGTH: 16 <212> TYPE: PRT <213> ORGANISM:
Artificial Sequence <220> FEATURE: <221> NAME/KEY:
source <223> OTHER INFORMATION: /note="Description of
Artificial Sequence: Synthetic peptide" <400> SEQUENCE: 7 Gln
Ser Ser Lys Ser Leu Leu His Ser Asn Gly Lys Thr Tyr Leu Asn 1 5 10
15 <210> SEQ ID NO 8 <211> LENGTH: 7 <212> TYPE:
PRT <213> ORGANISM: Artificial Sequence <220> FEATURE:
<221> NAME/KEY: source <223> OTHER INFORMATION:
/note="Description of Artificial Sequence: Synthetic peptide"
<400> SEQUENCE: 8 Trp Met Ser Thr Arg Ala Ser 1 5 <210>
SEQ ID NO 9 <211> LENGTH: 9 <212> TYPE: PRT <213>
ORGANISM: Artificial Sequence <220> FEATURE: <221>
NAME/KEY: source <223> OTHER INFORMATION: /note="Description
of Artificial Sequence: Synthetic peptide" <400> SEQUENCE: 9
Gln Gln Phe Leu Glu Phe Pro Phe Thr 1 5 <210> SEQ ID NO 10
<211> LENGTH: 118 <212> TYPE: PRT <213> ORGANISM:
Artificial Sequence <220> FEATURE: <221> NAME/KEY:
source <223> OTHER INFORMATION: /note="Description of
Artificial Sequence: Synthetic polypeptide" <400> SEQUENCE:
10 Glu Val Lys Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly
1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Thr
Asn Phe 20 25 30 Tyr Met Ser Trp Ile Arg Gln Pro Pro Gly Arg Ala
Pro Glu Trp Leu 35 40 45 Gly Val Ile Arg Asn Arg Pro Asn Gly Tyr
Thr Thr Asp Tyr Asn Pro 50 55 60 Ser Val Lys Gly Arg Phe Thr Ile
Ser Arg Asp Asn Thr Gln Asn Ile 65 70 75 80 Leu Tyr Leu Gln Met Ser
Thr Leu Arg Ala Asp Asp Thr Ala Phe Tyr 85 90 95 Tyr Cys Thr Arg
Leu Thr Tyr Gly Phe Asp Tyr Trp Gly Gln Gly Val 100 105 110 Met Val
Thr Val Ser Ser 115 <210> SEQ ID NO 11 <211> LENGTH:
112 <212> TYPE: PRT <213> ORGANISM: Artificial Sequence
<220> FEATURE: <221> NAME/KEY: source <223> OTHER
INFORMATION: /note="Description of Artificial Sequence: Synthetic
polypeptide" <400> SEQUENCE: 11 Asp Ile Val Met Thr Gln Gly
Ala Leu Pro Asn Pro Val Pro Ser Gly 1 5 10 15 Glu Ser Ala Ser Ile
Thr Cys Gln Ser Ser Lys Ser Leu Leu His Ser 20 25 30 Asn Gly Lys
Thr Tyr Leu Asn Trp Tyr Leu Gln Arg Pro Gly Gln Ser 35 40 45 Pro
Gln Leu Leu Ile Tyr Trp Met Ser Thr Arg Ala Ser Gly Val Ser 50 55
60 Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Lys Ile
65 70 75 80 Ser Ser Val Glu Ala Glu Val Val Gly Val Tyr Tyr Cys Gln
Gln Phe 85 90 95 Leu Glu Phe Pro Phe Thr Phe Gly Ser Gly Thr Lys
Leu Glu Ile Lys 100 105 110 <210> SEQ ID NO 12 <211>
LENGTH: 10 <212> TYPE: PRT <213> ORGANISM: Artificial
Sequence <220> FEATURE: <221> NAME/KEY: source
<223> OTHER INFORMATION: /note="Description of Artificial
Sequence: Synthetic peptide" <400> SEQUENCE: 12 Gly Phe Thr
Phe Thr Asn Phe Tyr Met Ser 1 5 10 <210> SEQ ID NO 13
<211> LENGTH: 19 <212> TYPE: PRT <213> ORGANISM:
Artificial Sequence <220> FEATURE: <221> NAME/KEY:
source <223> OTHER INFORMATION: /note="Description of
Artificial Sequence: Synthetic peptide" <400> SEQUENCE: 13
Val Ile Arg Asn Arg Pro Asn Gly Tyr Thr Thr Asp Tyr Asn Pro Ser 1 5
10 15 Val Lys Gly <210> SEQ ID NO 14 <211> LENGTH: 9
<212> TYPE: PRT <213> ORGANISM: Artificial Sequence
<220> FEATURE: <221> NAME/KEY: source <223> OTHER
INFORMATION: /note="Description of Artificial Sequence: Synthetic
peptide" <400> SEQUENCE: 14 Thr Arg Leu Thr Tyr Gly Phe Asp
Tyr 1 5 <210> SEQ ID NO 15 <211> LENGTH: 118
<212> TYPE: PRT <213> ORGANISM: Artificial Sequence
<220> FEATURE: <221> NAME/KEY: source <223> OTHER
INFORMATION: /note="Description of Artificial Sequence: Synthetic
polypeptide" <400> SEQUENCE: 15 Glu Val Lys Leu Leu Asp Ser
Gly Gly Gly Leu Val Gln Ala Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser
Cys Ala Gly Ser Gly Phe Thr Phe Thr Asp Phe 20 25 30 Tyr Met Ser
Trp Ile Arg Gln Pro Pro Gly Lys Ala Pro Glu Trp Leu 35 40 45 Gly
Val Ile Arg Asn Lys Ala Asn Gly Tyr Thr Ala Gly Tyr Asn Pro 50 55
60 Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Thr Gln Asn Ile
65 70 75 80 Leu Tyr Leu Gln Met Asn Thr Leu Arg Ala Glu Asp Thr Ala
Ile Tyr 85 90 95 Tyr Cys Ala Arg Leu Thr Tyr Gly Phe Asp Tyr Trp
Gly Gln Gly Val 100 105 110 Met Val Thr Val Ser Ser 115 <210>
SEQ ID NO 16 <211> LENGTH: 112 <212> TYPE: PRT
<213> ORGANISM: Artificial Sequence <220> FEATURE:
<221> NAME/KEY: source <223> OTHER INFORMATION:
/note="Description of Artificial Sequence: Synthetic polypeptide"
<400> SEQUENCE: 16 Asp Ile Val Met Thr Gln Gly Ala Leu Pro
Asn Pro Val Pro Ser Gly 1 5 10 15 Glu Ser Ala Ser Ile Thr Cys Gln
Ser Ser Lys Ser Leu Leu His Ser 20 25 30 Asn Gly Lys Thr Tyr Leu
Asn Trp Tyr Leu Gln Arg Pro Gly Gln Ser 35 40 45 Pro Gln Leu Leu
Ile Tyr Trp Met Ser Thr Arg Ala Ser Gly Val Ser 50 55 60 Asp Arg
Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Lys Ile 65 70 75 80
Ser Ser Val Glu Ala Glu Asp Val Gly Val Tyr Tyr Cys Gln Gln Phe 85
90 95 Leu Glu Tyr Pro Phe Thr Phe Gly Ser Gly Thr Lys Leu Glu Ile
Lys 100 105 110 <210> SEQ ID NO 17 <211> LENGTH: 9
<212> TYPE: PRT <213> ORGANISM: Artificial Sequence
<220> FEATURE: <221> NAME/KEY: source <223> OTHER
INFORMATION: /note="Description of Artificial Sequence: Synthetic
peptide" <400> SEQUENCE: 17 Ala Arg Leu Thr Tyr Gly Phe Asp
Tyr 1 5 <210> SEQ ID NO 18 <211> LENGTH: 9 <212>
TYPE: PRT <213> ORGANISM: Artificial Sequence <220>
FEATURE: <221> NAME/KEY: source <223> OTHER
INFORMATION: /note="Description of Artificial Sequence: Synthetic
peptide" <400> SEQUENCE: 18 Gln Gln Phe Leu Glu Tyr Pro Phe
Thr 1 5 <210> SEQ ID NO 19 <211> LENGTH: 118
<212> TYPE: PRT <213> ORGANISM: Artificial Sequence
<220> FEATURE: <221> NAME/KEY: source <223> OTHER
INFORMATION: /note="Description of Artificial Sequence: Synthetic
polypeptide" <400> SEQUENCE: 19 Glu Val Gln Leu Val Glu Ser
Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser
Cys Ala Ala Ser Gly Phe Thr Phe Thr Asp Phe 20 25 30 Tyr Met Ser
Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser
Val Ile Arg Asn Lys Ala Asn Gly Tyr Thr Ala Gly Tyr Asn Pro 50 55
60 Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr
65 70 75 80 Leu Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala
Val Tyr 85 90 95 Tyr Cys Ala Arg Leu Thr Tyr Gly Phe Asp Tyr Trp
Gly Gln Gly Thr 100 105 110 Leu Val Thr Val Ser Ser 115 <210>
SEQ ID NO 20 <211> LENGTH: 112 <212> TYPE: PRT
<213> ORGANISM: Artificial Sequence <220> FEATURE:
<221> NAME/KEY: source <223> OTHER INFORMATION:
/note="Description of Artificial Sequence: Synthetic polypeptide"
<400> SEQUENCE: 20 Asp Ile Val Met Thr Gln Thr Pro Leu Ser
Leu Pro Val Thr Pro Gly 1 5 10 15 Glu Pro Ala Ser Ile Ser Cys Gln
Ser Ser Lys Ser Leu Leu His Ser 20 25 30 Asn Gly Lys Thr Tyr Leu
Asn Trp Tyr Leu Gln Lys Pro Gly Gln Ser 35 40 45 Pro Gln Leu Leu
Ile Tyr Trp Met Ser Thr Arg Ala Ser Gly Val Pro 50 55 60 Asp Arg
Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Lys Ile 65 70 75 80
Ser Arg Val Glu Ala Glu Asp Val Gly Val Tyr Tyr Cys Gln Gln Phe 85
90 95 Leu Glu Tyr Pro Phe Thr Phe Gly Gln Gly Thr Lys Val Glu Ile
Lys 100 105 110 <210> SEQ ID NO 21 <211> LENGTH: 118
<212> TYPE: PRT <213> ORGANISM: Artificial Sequence
<220> FEATURE: <221> NAME/KEY: source <223> OTHER
INFORMATION: /note="Description of Artificial Sequence: Synthetic
polypeptide" <400> SEQUENCE: 21 Glu Val Gln Leu Val Glu Ser
Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser
Cys Ala Gly Ser Gly Phe Thr Phe Thr Asp Phe 20 25 30 Tyr Met Ser
Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser
Val Ile Arg Asn Lys Ala Asn Gly Tyr Thr Ala Gly Tyr Asn Pro 50 55
60 Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr
65 70 75 80 Leu Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala
Val Tyr 85 90 95 Tyr Cys Ala Arg Leu Thr Tyr Gly Phe Asp Tyr Trp
Gly Gln Gly Thr 100 105 110 Leu Val Thr Val Ser Ser 115 <210>
SEQ ID NO 22 <211> LENGTH: 112 <212> TYPE: PRT
<213> ORGANISM: Artificial Sequence <220> FEATURE:
<221> NAME/KEY: source <223> OTHER INFORMATION:
/note="Description of Artificial Sequence: Synthetic polypeptide"
<400> SEQUENCE: 22 Asp Ile Val Met Thr Gln Thr Pro Leu Ser
Leu Pro Val Thr Pro Gly 1 5 10 15 Glu Pro Ala Ser Ile Ser Cys Gln
Ser Ser Lys Ser Leu Leu His Ser 20 25 30 Thr Gly Lys Thr Tyr Leu
Asn Trp Tyr Leu Gln Lys Pro Gly Gln Ser 35 40 45 Pro Gln Leu Leu
Ile Tyr Trp Met Ser Thr Arg Ala Ser Gly Val Pro 50 55 60 Asp Arg
Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Lys Ile 65 70 75 80
Ser Arg Val Glu Ala Glu Asp Val Gly Val Tyr Tyr Cys Gln Gln Phe 85
90 95 Leu Glu Tyr Pro Phe Thr Phe Gly Gln Gly Thr Lys Val Glu Ile
Lys 100 105 110 <210> SEQ ID NO 23 <211> LENGTH: 16
<212> TYPE: PRT <213> ORGANISM: Artificial Sequence
<220> FEATURE: <221> NAME/KEY: source <223> OTHER
INFORMATION: /note="Description of Artificial Sequence: Synthetic
peptide" <400> SEQUENCE: 23 Gln Ser Ser Lys Ser Leu Leu His
Ser Thr Gly Lys Thr Tyr Leu Asn 1 5 10 15 <210> SEQ ID NO 24
<211> LENGTH: 118 <212> TYPE: PRT <213> ORGANISM:
Artificial Sequence <220> FEATURE: <221> NAME/KEY:
source <223> OTHER INFORMATION: /note="Description of
Artificial Sequence: Synthetic polypeptide" <400> SEQUENCE:
24 Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly
1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Thr
Asp Phe 20 25 30 Tyr Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly
Leu Glu Trp Val 35 40 45 Ser Val Ile Arg Asn Lys Ala Asn Ala Tyr
Thr Ala Gly Tyr Asn Pro 50 55 60 Ser Val Lys Gly Arg Phe Thr Ile
Ser Arg Asp Asn Ser Lys Asn Thr 65 70 75 80 Leu Tyr Leu Gln Met Asn
Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr 85 90 95 Tyr Cys Ala Arg
Leu Thr Tyr Gly Phe Asp Tyr Trp Gly Gln Gly Thr 100 105 110 Leu Val
Thr Val Ser Ser 115 <210> SEQ ID NO 25 <211> LENGTH: 19
<212> TYPE: PRT <213> ORGANISM: Artificial Sequence
<220> FEATURE: <221> NAME/KEY: source <223> OTHER
INFORMATION: /note="Description of Artificial Sequence: Synthetic
peptide" <400> SEQUENCE: 25 Val Ile Arg Asn Lys Ala Asn Ala
Tyr Thr Ala Gly Tyr Asn Pro Ser 1 5 10 15 Val Lys Gly <210>
SEQ ID NO 26 <211> LENGTH: 118 <212> TYPE: PRT
<213> ORGANISM: Artificial Sequence <220> FEATURE:
<221> NAME/KEY: source <223> OTHER INFORMATION:
/note="Description of Artificial Sequence: Synthetic polypeptide"
<400> SEQUENCE: 26 Glu Val Gln Leu Val Glu Ser Gly Gly Gly
Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Gly
Ser Gly Phe Thr Phe Thr Asp Phe 20 25 30 Tyr Met Ser Trp Val Arg
Gln Ala Pro Gly Lys Gly Pro Glu Trp Leu 35 40 45 Ser Val Ile Arg
Asn Lys Ala Asn Ala Tyr Thr Ala Gly Tyr Asn Pro 50 55 60 Ser Val
Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr 65 70 75 80
Leu Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr 85
90 95 Tyr Cys Ala Arg Leu Thr Tyr Gly Phe Asp Tyr Trp Gly Gln Gly
Thr 100 105 110 Leu Val Thr Val Ser Ser 115 <210> SEQ ID NO
27 <211> LENGTH: 112 <212> TYPE: PRT <213>
ORGANISM: Artificial Sequence <220> FEATURE: <221>
NAME/KEY: source <223> OTHER INFORMATION: /note="Description
of Artificial Sequence: Synthetic polypeptide" <400>
SEQUENCE: 27 Asp Ile Val Met Thr Gln Ser Pro Asp Ser Leu Ala Val
Ser Leu Gly 1 5 10 15 Glu Arg Ala Thr Ile Asn Cys Gln Ser Ser Lys
Ser Leu Leu His Ser 20 25 30 Asn Gly Lys Thr Tyr Leu Asn Trp Tyr
Gln Gln Lys Pro Gly Gln Pro 35 40 45 Pro Lys Leu Leu Ile Tyr Trp
Met Ser Thr Arg Ala Ser Gly Val Pro 50 55 60 Asp Arg Phe Ser Gly
Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile 65 70 75 80 Ser Ser Leu
Gln Ala Glu Asp Val Ala Val Tyr Tyr Cys Gln Gln Phe 85 90 95 Leu
Glu Phe Pro Phe Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys 100 105
110 <210> SEQ ID NO 28 <211> LENGTH: 10 <212>
TYPE: PRT <213> ORGANISM: Artificial Sequence <220>
FEATURE: <221> NAME/KEY: source <223> OTHER
INFORMATION: /note="Description of Artificial Sequence: Synthetic
peptide" <220> FEATURE: <221> NAME/KEY: VARIANT
<222> LOCATION: (6)..(6) <223> OTHER INFORMATION:
/replace="Asn" <220> FEATURE: <221> NAME/KEY: SITE
<222> LOCATION: (1)..(10) <223> OTHER INFORMATION:
/note="Variant residues given in the sequence have no preference
with respect to those in the annotations for variant positions"
<400> SEQUENCE: 28 Gly Phe Thr Phe Thr Asp Phe Tyr Met Ser 1
5 10 <210> SEQ ID NO 29 <211> LENGTH: 19 <212>
TYPE: PRT <213> ORGANISM: Artificial Sequence <220>
FEATURE: <221> NAME/KEY: source <223> OTHER
INFORMATION: /note="Description of Artificial Sequence: Synthetic
peptide" <220> FEATURE: <221> NAME/KEY: VARIANT
<222> LOCATION: (5)..(5) <223> OTHER INFORMATION:
/replace="Arg" <220> FEATURE: <221> NAME/KEY: VARIANT
<222> LOCATION: (6)..(6) <223> OTHER INFORMATION:
/replace="Pro" <220> FEATURE: <221> NAME/KEY: VARIANT
<222> LOCATION: (8)..(8) <223> OTHER INFORMATION:
/replace="Ala" <220> FEATURE: <221> NAME/KEY: VARIANT
<222> LOCATION: (11)..(11) <223> OTHER INFORMATION:
/replace="Thr" <220> FEATURE: <221> NAME/KEY: VARIANT
<222> LOCATION: (12)..(12) <223> OTHER INFORMATION:
/replace="Asp" <220> FEATURE: <221> NAME/KEY: SITE
<222> LOCATION: (1)..(19) <223> OTHER INFORMATION:
/note="Variant residues given in the sequence have no preference
with respect to those in the annotations for variant positions"
<400> SEQUENCE: 29 Val Ile Arg Asn Lys Ala Asn Gly Tyr Thr
Ala Gly Tyr Asn Pro Ser 1 5 10 15 Val Lys Gly <210> SEQ ID NO
30 <211> LENGTH: 9 <212> TYPE: PRT <213>
ORGANISM: Artificial Sequence <220> FEATURE: <221>
NAME/KEY: source <223> OTHER INFORMATION: /note="Description
of Artificial Sequence: Synthetic peptide" <220> FEATURE:
<221> NAME/KEY: VARIANT <222> LOCATION: (1)..(1)
<223> OTHER INFORMATION: /replace="Thr" <220> FEATURE:
<221> NAME/KEY: VARIANT <222> LOCATION: (4)..(4)
<223> OTHER INFORMATION: /replace="Ser" <220> FEATURE:
<221> NAME/KEY: SITE <222> LOCATION: (1)..(9)
<223> OTHER INFORMATION: /note="Variant residues given in the
sequence have no preference with respect to those in the
annotations for variant positions" <400> SEQUENCE: 30 Ala Arg
Leu Thr Tyr Gly Phe Asp Tyr 1 5 <210> SEQ ID NO 31
<211> LENGTH: 16 <212> TYPE: PRT <213> ORGANISM:
Artificial Sequence <220> FEATURE: <221> NAME/KEY:
source <223> OTHER INFORMATION: /note="Description of
Artificial Sequence: Synthetic peptide" <220> FEATURE:
<221> NAME/KEY: VARIANT <222> LOCATION: (10)..(10)
<223> OTHER INFORMATION: /replace="Thr" <220> FEATURE:
<221> NAME/KEY: SITE <222> LOCATION: (1)..(16)
<223> OTHER INFORMATION: /note="Variant residues given in the
sequence have no preference with respect to those in the
annotations for variant positions" <400> SEQUENCE: 31 Gln Ser
Ser Lys Ser Leu Leu His Ser Asn Gly Lys Thr Tyr Leu Asn 1 5 10 15
<210> SEQ ID NO 32 <211> LENGTH: 9 <212> TYPE:
PRT <213> ORGANISM: Artificial Sequence <220> FEATURE:
<221> NAME/KEY: source <223> OTHER INFORMATION:
/note="Description of Artificial Sequence: Synthetic peptide"
<220> FEATURE: <221> NAME/KEY: VARIANT <222>
LOCATION: (6)..(6) <223> OTHER INFORMATION: /replace="Phe"
<220> FEATURE: <221> NAME/KEY: SITE <222>
LOCATION: (1)..(9) <223> OTHER INFORMATION: /note="Variant
residues given in the sequence have no preference with respect to
those in the annotations for variant positions" <400>
SEQUENCE: 32 Gln Gln Phe Leu Glu Tyr Pro Phe Thr 1 5 <210>
SEQ ID NO 33 <400> SEQUENCE: 33 000 <210> SEQ ID NO 34
<211> LENGTH: 5 <212> TYPE: PRT <213> ORGANISM:
Artificial Sequence <220> FEATURE: <221> NAME/KEY:
source <223> OTHER INFORMATION: /note="Description of
Artificial Sequence: Synthetic peptide" <400> SEQUENCE: 34
Gly Gly Gly Gly Ser 1 5 <210> SEQ ID NO 35 <211>
LENGTH: 6 <212> TYPE: PRT <213> ORGANISM: Artificial
Sequence <220> FEATURE: <221> NAME/KEY: source
<223> OTHER INFORMATION: /note="Description of Artificial
Sequence: Synthetic 6xHis tag" <400> SEQUENCE: 35 His His His
His His His 1 5 <210> SEQ ID NO 36 <211> LENGTH: 113
<212> TYPE: PRT <213> ORGANISM: Artificial Sequence
<220> FEATURE: <221> NAME/KEY: source <223> OTHER
INFORMATION: /note="Description of Artificial Sequence: Synthetic
polypeptide" <400> SEQUENCE: 36 Ala Ser Thr Lys Gly Pro Ser
Val Phe Pro Leu Ala Pro Ser Ser Lys 1 5 10 15 Ser Thr Ser Gly Gly
Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr 20 25 30 Phe Pro Glu
Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser 35 40 45 Gly
Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser 50 55
60 Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr
65 70 75 80 Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val
Asp Lys 85 90 95 Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr
Cys Pro Pro Cys 100 105 110 Pro <210> SEQ ID NO 37
<211> LENGTH: 107 <212> TYPE: PRT <213> ORGANISM:
Artificial Sequence <220> FEATURE: <221> NAME/KEY:
source <223> OTHER INFORMATION: /note="Description of
Artificial Sequence: Synthetic polypeptide" <400> SEQUENCE:
37 Arg Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu
1 5 10 15 Gln Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn
Asn Phe 20 25 30 Tyr Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp
Asn Ala Leu Gln 35 40 45 Ser Gly Asn Ser Gln Glu Ser Val Thr Glu
Gln Asp Ser Lys Asp Ser 50 55 60 Thr Tyr Ser Leu Ser Ser Thr Leu
Thr Leu Ser Lys Ala Asp Tyr Glu 65 70 75 80 Lys His Lys Val Tyr Ala
Cys Glu Val Thr His Gln Gly Leu Ser Ser 85 90 95 Pro Val Thr Lys
Ser Phe Asn Arg Gly Glu Cys 100 105 <210> SEQ ID NO 38
<211> LENGTH: 217 <212> TYPE: PRT <213> ORGANISM:
Homo sapiens <400> SEQUENCE: 38 Ala Pro Glu Leu Leu Gly Gly
Pro Ser Val Phe Leu Phe Pro Pro Lys 1 5 10 15 Pro Lys Asp Thr Leu
Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val 20 25 30 Val Val Asp
Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr 35 40 45 Val
Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu 50 55
60 Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His
65 70 75 80 Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser
Asn Lys 85 90 95 Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys
Ala Lys Gly Gln 100 105 110 Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro
Pro Ser Arg Asp Glu Leu 115 120 125 Thr Lys Asn Gln Val Ser Leu Thr
Cys Leu Val Lys Gly Phe Tyr Pro 130 135 140 Ser Asp Ile Ala Val Glu
Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn 145 150 155 160 Tyr Lys Thr
Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu 165 170 175 Tyr
Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val 180 185
190 Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln
195 200 205 Lys Ser Leu Ser Leu Ser Pro Gly Lys 210 215 <210>
SEQ ID NO 39 <211> LENGTH: 217 <212> TYPE: PRT
<213> ORGANISM: Artificial Sequence <220> FEATURE:
<221> NAME/KEY: source <223> OTHER INFORMATION:
/note="Description of Artificial Sequence: Synthetic polypeptide"
<400> SEQUENCE: 39 Ala Pro Glu Ala Ala Gly Gly Pro Ser Val
Phe Leu Phe Pro Pro Lys 1 5 10 15 Pro Lys Asp Thr Leu Met Ile Ser
Arg Thr Pro Glu Val Thr Cys Val 20 25 30 Val Val Asp Val Ser His
Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr 35 40 45 Val Asp Gly Val
Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu 50 55 60 Gln Tyr
Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His 65 70 75 80
Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys 85
90 95 Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly
Gln 100 105 110 Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg
Asp Glu Leu 115 120 125 Thr Lys Asn Gln Val Ser Leu Ser Cys Ala Val
Lys Gly Phe Tyr Pro 130 135 140 Ser Asp Ile Ala Val Glu Trp Glu Ser
Asn Gly Gln Pro Glu Asn Asn 145 150 155 160 Tyr Lys Thr Thr Pro Pro
Val Leu Asp Ser Asp Gly Ser Phe Phe Leu 165 170 175 Val Ser Lys Leu
Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val 180 185 190 Phe Ser
Cys Ser Val Leu His Glu Ala Leu His Ser His Tyr Thr Gln 195 200 205
Lys Ser Leu Ser Leu Ser Pro Gly Lys 210 215 <210> SEQ ID NO
40 <211> LENGTH: 217 <212> TYPE: PRT <213>
ORGANISM: Artificial Sequence <220> FEATURE: <221>
NAME/KEY: source <223> OTHER INFORMATION: /note="Description
of Artificial Sequence: Synthetic polypeptide" <400>
SEQUENCE: 40 Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe
Pro Pro Lys 1 5 10 15 Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro
Glu Val Thr Cys Val 20 25 30 Val Val Asp Val Ser His Glu Asp Pro
Glu Val Lys Phe Asn Trp Tyr 35 40 45 Val Asp Gly Val Glu Val His
Asn Ala Lys Thr Lys Pro Arg Glu Glu 50 55 60 Gln Tyr Asn Ser Thr
Tyr Arg Val Val Ser Val Leu Thr Val Leu His 65 70 75 80 Gln Asp Trp
Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys 85 90 95 Ala
Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln 100 105
110 Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu
115 120 125 Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe
Tyr Pro 130 135 140 Ser Asp Ile Ala Val Trp Trp Glu Ser Tyr Gly Thr
Glu Trp Ser Ser 145 150 155 160 Tyr Lys Thr Thr Pro Pro Val Leu Asp
Ser Asp Gly Ser Phe Phe Leu 165 170 175 Tyr Ser Lys Leu Thr Val Thr
Lys Glu Glu Trp Gln Gln Gly Phe Val 180 185 190 Phe Ser Cys Ser Val
Met His Glu Ala Leu His Asn His Tyr Thr Gln 195 200 205 Lys Ser Leu
Ser Leu Ser Pro Gly Lys 210 215 <210> SEQ ID NO 41
<211> LENGTH: 217 <212> TYPE: PRT <213> ORGANISM:
Artificial Sequence <220> FEATURE: <221> NAME/KEY:
source <223> OTHER INFORMATION: /note="Description of
Artificial Sequence: Synthetic polypeptide" <400> SEQUENCE:
41 Ala Pro Glu Ala Ala Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys
1 5 10 15 Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr
Cys Val 20 25 30 Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys
Phe Asn Trp Tyr 35 40 45 Val Asp Gly Val Glu Val His Asn Ala Lys
Thr Lys Pro Arg Glu Glu 50 55 60 Gln Tyr Asn Ser Thr Tyr Arg Val
Val Ser Val Leu Thr Val Leu His 65 70 75 80 Gln Asp Trp Leu Asn Gly
Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys 85 90 95 Ala Leu Pro Ala
Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln 100 105 110 Pro Arg
Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu 115 120 125
Thr Lys Asn Gln Val Ser Leu Trp Cys Leu Val Lys Gly Phe Tyr Pro 130
135 140 Ser Asp Ile Ala Val Trp Trp Glu Ser Tyr Gly Thr Glu Trp Ser
Ser 145 150 155 160 Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly
Ser Phe Phe Leu 165 170 175 Tyr Ser Lys Leu Thr Val Thr Lys Glu Glu
Trp Gln Gln Gly Phe Val 180 185 190 Phe Ser Cys Ser Val Leu His Glu
Ala Leu His Ser His Tyr Thr Gln 195 200 205 Lys Ser Leu Ser Leu Ser
Pro Gly Lys 210 215 <210> SEQ ID NO 42 <211> LENGTH:
448 <212> TYPE: PRT <213> ORGANISM: Artificial Sequence
<220> FEATURE: <221> NAME/KEY: source <223> OTHER
INFORMATION: /note="Description of Artificial Sequence: Synthetic
polypeptide" <400> SEQUENCE: 42 Glu Val Gln Leu Val Glu Ser
Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser
Cys Ala Ala Ser Gly Phe Thr Phe Thr Asp Phe 20 25 30 Tyr Met Ser
Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser
Val Ile Arg Asn Lys Ala Asn Ala Tyr Thr Ala Gly Tyr Asn Pro 50 55
60 Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr
65 70 75 80 Leu Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala
Val Tyr 85 90 95 Tyr Cys Ala Arg Leu Thr Tyr Gly Phe Asp Tyr Trp
Gly Gln Gly Thr 100 105 110 Leu Val Thr Val Ser Ser Ala Ser Thr Lys
Gly Pro Ser Val Phe Pro 115 120 125 Leu Ala Pro Ser Ser Lys Ser Thr
Ser Gly Gly Thr Ala Ala Leu Gly 130 135 140 Cys Leu Val Lys Asp Tyr
Phe Pro Glu Pro Val Thr Val Ser Trp Asn 145 150 155 160 Ser Gly Ala
Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln 165 170 175 Ser
Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser 180 185
190 Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser
195 200 205 Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp
Lys Thr 210 215 220 His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala
Gly Gly Pro Ser 225 230 235 240 Val Phe Leu Phe Pro Pro Lys Pro Lys
Asp Thr Leu Met Ile Ser Arg 245 250 255 Thr Pro Glu Val Thr Cys Val
Val Val Asp Val Ser His Glu Asp Pro 260 265 270 Glu Val Lys Phe Asn
Trp Tyr Val Asp Gly Val Glu Val His Asn Ala 275 280 285 Lys Thr Lys
Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val 290 295 300 Ser
Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr 305 310
315 320 Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys
Thr 325 330 335 Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val
Tyr Thr Leu 340 345 350 Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln
Val Ser Leu Trp Cys 355 360 365 Leu Val Lys Gly Phe Tyr Pro Ser Asp
Ile Ala Val Trp Trp Glu Ser 370 375 380 Tyr Gly Thr Glu Trp Ser Ser
Tyr Lys Thr Thr Pro Pro Val Leu Asp 385 390 395 400 Ser Asp Gly Ser
Phe Phe Leu Tyr Ser Lys Leu Thr Val Thr Lys Glu 405 410 415 Glu Trp
Gln Gln Gly Phe Val Phe Ser Cys Ser Val Leu His Glu Ala 420 425 430
Leu His Ser His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys 435
440 445 <210> SEQ ID NO 43 <211> LENGTH: 217
<212> TYPE: PRT <213> ORGANISM: Artificial Sequence
<220> FEATURE: <221> NAME/KEY: source <223> OTHER
INFORMATION: /note="Description of Artificial Sequence: Synthetic
polypeptide" <400> SEQUENCE: 43 Ala Pro Glu Leu Leu Gly Gly
Pro Ser Val Phe Leu Phe Pro Pro Lys 1 5 10 15 Pro Lys Asp Thr Leu
Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val 20 25 30 Val Val Asp
Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr 35 40 45 Val
Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu 50 55
60 Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His
65 70 75 80 Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser
Asn Lys 85 90 95 Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys
Ala Lys Gly Gln 100 105 110 Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro
Pro Ser Arg Asp Glu Leu 115 120 125 Thr Lys Asn Gln Val Ser Leu Thr
Cys Leu Val Lys Gly Phe Tyr Pro 130 135 140 Ser Asp Ile Ala Val Glu
Trp Glu Ser Phe Gly Thr Glu Trp Ser Asn 145 150 155 160 Tyr Lys Thr
Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu 165 170 175 Tyr
Ser Lys Leu Thr Val Ser Lys Glu Glu Trp Gln Gln Gly Phe Val 180 185
190 Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln
195 200 205 Lys Ser Leu Ser Leu Ser Pro Gly Lys 210 215 <210>
SEQ ID NO 44 <211> LENGTH: 217 <212> TYPE: PRT
<213> ORGANISM: Artificial Sequence <220> FEATURE:
<221> NAME/KEY: source <223> OTHER INFORMATION:
/note="Description of Artificial Sequence: Synthetic polypeptide"
<400> SEQUENCE: 44 Ala Pro Glu Ala Ala Gly Gly Pro Ser Val
Phe Leu Phe Pro Pro Lys 1 5 10 15 Pro Lys Asp Thr Leu Met Ile Ser
Arg Thr Pro Glu Val Thr Cys Val 20 25 30 Val Val Asp Val Ser His
Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr 35 40 45 Val Asp Gly Val
Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu 50 55 60 Gln Tyr
Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His 65 70 75 80
Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys 85
90 95 Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly
Gln 100 105 110 Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg
Asp Glu Leu 115 120 125 Thr Lys Asn Gln Val Ser Leu Trp Cys Leu Val
Lys Gly Phe Tyr Pro 130 135 140 Ser Asp Ile Ala Val Glu Trp Glu Ser
Phe Gly Thr Glu Trp Ser Asn 145 150 155 160 Tyr Lys Thr Thr Pro Pro
Val Leu Asp Ser Asp Gly Ser Phe Phe Leu 165 170 175 Tyr Ser Lys Leu
Thr Val Ser Lys Glu Glu Trp Gln Gln Gly Phe Val 180 185 190 Phe Ser
Cys Ser Val Leu His Glu Ala Leu His Ser His Tyr Thr Gln 195 200 205
Lys Ser Leu Ser Leu Ser Pro Gly Lys 210 215 <210> SEQ ID NO
45 <211> LENGTH: 448 <212> TYPE: PRT <213>
ORGANISM: Artificial Sequence <220> FEATURE: <221>
NAME/KEY: source <223> OTHER INFORMATION: /note="Description
of Artificial Sequence: Synthetic polypeptide" <400>
SEQUENCE: 45 Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln
Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe
Thr Phe Thr Asp Phe 20 25 30 Tyr Met Ser Trp Val Arg Gln Ala Pro
Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Val Ile Arg Asn Lys Ala
Asn Ala Tyr Thr Ala Gly Tyr Asn Pro 50 55 60 Ser Val Lys Gly Arg
Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr 65 70 75 80 Leu Tyr Leu
Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr 85 90 95 Tyr
Cys Ala Arg Leu Thr Tyr Gly Phe Asp Tyr Trp Gly Gln Gly Thr 100 105
110 Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro
115 120 125 Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala
Leu Gly 130 135 140 Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr
Val Ser Trp Asn 145 150 155 160 Ser Gly Ala Leu Thr Ser Gly Val His
Thr Phe Pro Ala Val Leu Gln 165 170 175 Ser Ser Gly Leu Tyr Ser Leu
Ser Ser Val Val Thr Val Pro Ser Ser 180 185 190 Ser Leu Gly Thr Gln
Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser 195 200 205 Asn Thr Lys
Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr 210 215 220 His
Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Gly Pro Ser 225 230
235 240 Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser
Arg 245 250 255 Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His
Glu Asp Pro 260 265 270 Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val
Glu Val His Asn Ala 275 280 285 Lys Thr Lys Pro Arg Glu Glu Gln Tyr
Asn Ser Thr Tyr Arg Val Val 290 295 300 Ser Val Leu Thr Val Leu His
Gln Asp Trp Leu Asn Gly Lys Glu Tyr 305 310 315 320 Lys Cys Lys Val
Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr 325 330 335 Ile Ser
Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu 340 345 350
Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Trp Cys 355
360 365 Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu
Ser 370 375 380 Phe Gly Thr Glu Trp Ser Asn Tyr Lys Thr Thr Pro Pro
Val Leu Asp 385 390 395 400 Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys
Leu Thr Val Ser Lys Glu 405 410 415 Glu Trp Gln Gln Gly Phe Val Phe
Ser Cys Ser Val Leu His Glu Ala 420 425 430 Leu His Ser His Tyr Thr
Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys 435 440 445 <210> SEQ
ID NO 46 <211> LENGTH: 217 <212> TYPE: PRT <213>
ORGANISM: Artificial Sequence <220> FEATURE: <221>
NAME/KEY: source <223> OTHER INFORMATION: /note="Description
of Artificial Sequence: Synthetic polypeptide" <400>
SEQUENCE: 46 Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe
Pro Pro Lys 1 5 10 15 Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro
Glu Val Thr Cys Val 20 25 30 Val Val Asp Val Ser His Glu Asp Pro
Glu Val Lys Phe Asn Trp Tyr 35 40 45 Val Asp Gly Val Glu Val His
Asn Ala Lys Thr Lys Pro Arg Glu Glu 50 55 60 Gln Tyr Asn Ser Thr
Tyr Arg Val Val Ser Val Leu Thr Val Leu His 65 70 75 80 Gln Asp Trp
Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys 85 90 95 Ala
Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln 100 105
110 Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu
115 120 125 Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe
Tyr Pro 130 135 140 Ser Asp Ile Ala Val Glu Trp Glu Ser Tyr Gly Thr
Glu Trp Val Asn 145 150 155 160 Tyr Lys Thr Thr Pro Pro Val Leu Asp
Ser Asp Gly Ser Phe Phe Leu 165 170 175 Tyr Ser Lys Leu Thr Val Thr
Lys Glu Glu Trp Gln Gln Gly Phe Val 180 185 190 Phe Ser Cys Ser Val
Met His Glu Ala Leu His Asn His Tyr Thr Gln 195 200 205 Lys Ser Leu
Ser Leu Ser Pro Gly Lys 210 215 <210> SEQ ID NO 47
<211> LENGTH: 217 <212> TYPE: PRT <213> ORGANISM:
Artificial Sequence <220> FEATURE: <221> NAME/KEY:
source <223> OTHER INFORMATION: /note="Description of
Artificial Sequence: Synthetic polypeptide" <400> SEQUENCE:
47 Ala Pro Glu Ala Ala Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys
1 5 10 15 Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr
Cys Val 20 25 30 Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys
Phe Asn Trp Tyr 35 40 45 Val Asp Gly Val Glu Val His Asn Ala Lys
Thr Lys Pro Arg Glu Glu 50 55 60 Gln Tyr Asn Ser Thr Tyr Arg Val
Val Ser Val Leu Thr Val Leu His 65 70 75 80 Gln Asp Trp Leu Asn Gly
Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys 85 90 95 Ala Leu Pro Ala
Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln 100 105 110 Pro Arg
Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu 115 120 125
Thr Lys Asn Gln Val Ser Leu Trp Cys Leu Val Lys Gly Phe Tyr Pro 130
135 140 Ser Asp Ile Ala Val Glu Trp Glu Ser Tyr Gly Thr Glu Trp Val
Asn 145 150 155 160 Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly
Ser Phe Phe Leu 165 170 175 Tyr Ser Lys Leu Thr Val Thr Lys Glu Glu
Trp Gln Gln Gly Phe Val 180 185 190 Phe Ser Cys Ser Val Leu His Glu
Ala Leu His Ser His Tyr Thr Gln 195 200 205 Lys Ser Leu Ser Leu Ser
Pro Gly Lys 210 215 <210> SEQ ID NO 48 <211> LENGTH:
448 <212> TYPE: PRT <213> ORGANISM: Artificial Sequence
<220> FEATURE: <221> NAME/KEY: source <223> OTHER
INFORMATION: /note="Description of Artificial Sequence: Synthetic
polypeptide" <400> SEQUENCE: 48 Glu Val Gln Leu Val Glu Ser
Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser
Cys Ala Ala Ser Gly Phe Thr Phe Thr Asp Phe 20 25 30 Tyr Met Ser
Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser
Val Ile Arg Asn Lys Ala Asn Ala Tyr Thr Ala Gly Tyr Asn Pro 50 55
60 Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr
65 70 75 80 Leu Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala
Val Tyr 85 90 95 Tyr Cys Ala Arg Leu Thr Tyr Gly Phe Asp Tyr Trp
Gly Gln Gly Thr 100 105 110 Leu Val Thr Val Ser Ser Ala Ser Thr Lys
Gly Pro Ser Val Phe Pro 115 120 125 Leu Ala Pro Ser Ser Lys Ser Thr
Ser Gly Gly Thr Ala Ala Leu Gly 130 135 140 Cys Leu Val Lys Asp Tyr
Phe Pro Glu Pro Val Thr Val Ser Trp Asn 145 150 155 160 Ser Gly Ala
Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln 165 170 175 Ser
Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser 180 185
190 Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser
195 200 205 Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp
Lys Thr 210 215 220 His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala
Gly Gly Pro Ser 225 230 235 240 Val Phe Leu Phe Pro Pro Lys Pro Lys
Asp Thr Leu Met Ile Ser Arg 245 250 255 Thr Pro Glu Val Thr Cys Val
Val Val Asp Val Ser His Glu Asp Pro 260 265 270 Glu Val Lys Phe Asn
Trp Tyr Val Asp Gly Val Glu Val His Asn Ala 275 280 285 Lys Thr Lys
Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val 290 295 300 Ser
Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr 305 310
315 320 Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys
Thr 325 330 335 Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val
Tyr Thr Leu 340 345 350 Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln
Val Ser Leu Trp Cys 355 360 365 Leu Val Lys Gly Phe Tyr Pro Ser Asp
Ile Ala Val Glu Trp Glu Ser 370 375 380 Tyr Gly Thr Glu Trp Val Asn
Tyr Lys Thr Thr Pro Pro Val Leu Asp 385 390 395 400 Ser Asp Gly Ser
Phe Phe Leu Tyr Ser Lys Leu Thr Val Thr Lys Glu 405 410 415 Glu Trp
Gln Gln Gly Phe Val Phe Ser Cys Ser Val Leu His Glu Ala 420 425 430
Leu His Ser His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys 435
440 445 <210> SEQ ID NO 49 <211> LENGTH: 217
<212> TYPE: PRT <213> ORGANISM: Artificial Sequence
<220> FEATURE: <221> NAME/KEY: source <223> OTHER
INFORMATION: /note="Description of Artificial Sequence: Synthetic
polypeptide" <400> SEQUENCE: 49 Ala Pro Glu Leu Leu Gly Gly
Pro Ser Val Phe Leu Phe Pro Pro Lys 1 5 10 15 Pro Lys Asp Thr Leu
Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val 20 25 30 Val Val Asp
Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr 35 40 45 Val
Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu 50 55
60 Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His
65 70 75 80 Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser
Asn Lys 85 90 95 Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys
Ala Lys Gly Gln 100 105 110 Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro
Pro Ser Arg Asp Glu Leu 115 120 125 Thr Lys Asn Gln Val Ser Leu Thr
Cys Leu Val Lys Gly Phe Tyr Pro 130 135 140 Ser Asp Ile Ala Val Glu
Trp Glu Ser Tyr Gly Thr Glu Trp Ser Asn 145 150 155 160 Tyr Lys Thr
Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu 165 170 175 Tyr
Ser Lys Leu Thr Val Ser Lys Glu Glu Trp Gln Gln Gly Phe Val 180 185
190 Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln
195 200 205 Lys Ser Leu Ser Leu Ser Pro Gly Lys 210 215 <210>
SEQ ID NO 50 <211> LENGTH: 217 <212> TYPE: PRT
<213> ORGANISM: Artificial Sequence <220> FEATURE:
<221> NAME/KEY: source <223> OTHER INFORMATION:
/note="Description of Artificial Sequence: Synthetic polypeptide"
<400> SEQUENCE: 50 Ala Pro Glu Ala Ala Gly Gly Pro Ser Val
Phe Leu Phe Pro Pro Lys 1 5 10 15 Pro Lys Asp Thr Leu Met Ile Ser
Arg Thr Pro Glu Val Thr Cys Val 20 25 30 Val Val Asp Val Ser His
Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr 35 40 45 Val Asp Gly Val
Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu 50 55 60 Gln Tyr
Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His 65 70 75 80
Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys 85
90 95 Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly
Gln 100 105 110 Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg
Asp Glu Leu 115 120 125 Thr Lys Asn Gln Val Ser Leu Trp Cys Leu Val
Lys Gly Phe Tyr Pro 130 135 140 Ser Asp Ile Ala Val Glu Trp Glu Ser
Tyr Gly Thr Glu Trp Ser Asn 145 150 155 160 Tyr Lys Thr Thr Pro Pro
Val Leu Asp Ser Asp Gly Ser Phe Phe Leu 165 170 175 Tyr Ser Lys Leu
Thr Val Ser Lys Glu Glu Trp Gln Gln Gly Phe Val 180 185 190 Phe Ser
Cys Ser Val Leu His Glu Ala Leu His Ser His Tyr Thr Gln 195 200 205
Lys Ser Leu Ser Leu Ser Pro Gly Lys 210 215 <210> SEQ ID NO
51 <211> LENGTH: 448 <212> TYPE: PRT <213>
ORGANISM: Artificial Sequence <220> FEATURE: <221>
NAME/KEY: source <223> OTHER INFORMATION: /note="Description
of Artificial Sequence: Synthetic polypeptide" <400>
SEQUENCE: 51 Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln
Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe
Thr Phe Thr Asp Phe 20 25 30 Tyr Met Ser Trp Val Arg Gln Ala Pro
Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Val Ile Arg Asn Lys Ala
Asn Ala Tyr Thr Ala Gly Tyr Asn Pro 50 55 60 Ser Val Lys Gly Arg
Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr 65 70 75 80 Leu Tyr Leu
Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr 85 90 95 Tyr
Cys Ala Arg Leu Thr Tyr Gly Phe Asp Tyr Trp Gly Gln Gly Thr 100 105
110 Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro
115 120 125 Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala
Leu Gly 130 135 140 Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr
Val Ser Trp Asn 145 150 155 160 Ser Gly Ala Leu Thr Ser Gly Val His
Thr Phe Pro Ala Val Leu Gln 165 170 175 Ser Ser Gly Leu Tyr Ser Leu
Ser Ser Val Val Thr Val Pro Ser Ser 180 185 190 Ser Leu Gly Thr Gln
Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser 195 200 205 Asn Thr Lys
Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr 210 215 220 His
Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Gly Pro Ser 225 230
235 240 Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser
Arg 245 250 255 Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His
Glu Asp Pro 260 265 270 Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val
Glu Val His Asn Ala 275 280 285 Lys Thr Lys Pro Arg Glu Glu Gln Tyr
Asn Ser Thr Tyr Arg Val Val 290 295 300 Ser Val Leu Thr Val Leu His
Gln Asp Trp Leu Asn Gly Lys Glu Tyr 305 310 315 320 Lys Cys Lys Val
Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr 325 330 335 Ile Ser
Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu 340 345 350
Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Trp Cys 355
360 365 Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu
Ser 370 375 380 Tyr Gly Thr Glu Trp Ser Asn Tyr Lys Thr Thr Pro Pro
Val Leu Asp 385 390 395 400 Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys
Leu Thr Val Ser Lys Glu 405 410 415 Glu Trp Gln Gln Gly Phe Val Phe
Ser Cys Ser Val Leu His Glu Ala 420 425 430 Leu His Ser His Tyr Thr
Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys 435 440 445 <210> SEQ
ID NO 52 <211> LENGTH: 448 <212> TYPE: PRT <213>
ORGANISM: Artificial Sequence <220> FEATURE: <221>
NAME/KEY: source <223> OTHER INFORMATION: /note="Description
of Artificial Sequence: Synthetic polypeptide" <400>
SEQUENCE: 52 Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln
Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe
Thr Phe Thr Asp Phe 20 25 30 Tyr Met Ser Trp Val Arg Gln Ala Pro
Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Val Ile Arg Asn Lys Ala
Asn Ala Tyr Thr Ala Gly Tyr Asn Pro 50 55 60 Ser Val Lys Gly Arg
Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr 65 70 75 80 Leu Tyr Leu
Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr 85 90 95 Tyr
Cys Ala Arg Leu Thr Tyr Gly Phe Asp Tyr Trp Gly Gln Gly Thr 100 105
110 Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro
115 120 125 Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala
Leu Gly 130 135 140 Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr
Val Ser Trp Asn 145 150 155 160 Ser Gly Ala Leu Thr Ser Gly Val His
Thr Phe Pro Ala Val Leu Gln 165 170 175 Ser Ser Gly Leu Tyr Ser Leu
Ser Ser Val Val Thr Val Pro Ser Ser 180 185 190 Ser Leu Gly Thr Gln
Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser 195 200 205 Asn Thr Lys
Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr 210 215 220 His
Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser 225 230
235 240 Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser
Arg 245 250 255 Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His
Glu Asp Pro 260 265 270 Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val
Glu Val His Asn Ala 275 280 285 Lys Thr Lys Pro Arg Glu Glu Gln Tyr
Asn Ser Thr Tyr Arg Val Val 290 295 300 Ser Val Leu Thr Val Leu His
Gln Asp Trp Leu Asn Gly Lys Glu Tyr 305 310 315 320 Lys Cys Lys Val
Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr 325 330 335 Ile Ser
Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu 340 345 350
Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Ser Cys 355
360 365 Ala Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu
Ser 370 375 380 Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro
Val Leu Asp 385 390 395 400 Ser Asp Gly Ser Phe Phe Leu Val Ser Lys
Leu Thr Val Asp Lys Ser 405 410 415 Arg Trp Gln Gln Gly Asn Val Phe
Ser Cys Ser Val Leu His Glu Ala 420 425 430 Leu His Ser His Tyr Thr
Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys 435 440 445 <210> SEQ
ID NO 53 <211> LENGTH: 448 <212> TYPE: PRT <213>
ORGANISM: Artificial Sequence <220> FEATURE: <221>
NAME/KEY: source <223> OTHER INFORMATION: /note="Description
of Artificial Sequence: Synthetic polypeptide" <400>
SEQUENCE: 53 Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln
Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe
Thr Phe Thr Asp Phe 20 25 30 Tyr Met Ser Trp Val Arg Gln Ala Pro
Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Val Ile Arg Asn Lys Ala
Asn Ala Tyr Thr Ala Gly Tyr Asn Pro 50 55 60 Ser Val Lys Gly Arg
Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr 65 70 75 80 Leu Tyr Leu
Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr 85 90 95 Tyr
Cys Ala Arg Leu Thr Tyr Gly Phe Asp Tyr Trp Gly Gln Gly Thr 100 105
110 Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro
115 120 125 Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala
Leu Gly 130 135 140 Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr
Val Ser Trp Asn 145 150 155 160 Ser Gly Ala Leu Thr Ser Gly Val His
Thr Phe Pro Ala Val Leu Gln 165 170 175 Ser Ser Gly Leu Tyr Ser Leu
Ser Ser Val Val Thr Val Pro Ser Ser 180 185 190 Ser Leu Gly Thr Gln
Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser 195 200 205 Asn Thr Lys
Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr 210 215 220 His
Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Gly Pro Ser 225 230
235 240 Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser
Arg 245 250 255 Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His
Glu Asp Pro 260 265 270 Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val
Glu Val His Asn Ala 275 280 285 Lys Thr Lys Pro Arg Glu Glu Gln Tyr
Asn Ser Thr Tyr Arg Val Val 290 295 300 Ser Val Leu Thr Val Leu His
Gln Asp Trp Leu Asn Gly Lys Glu Tyr 305 310 315 320 Lys Cys Lys Val
Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr 325 330 335 Ile Ser
Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu 340 345 350
Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Ser Cys 355
360 365 Ala Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu
Ser 370 375 380 Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro
Val Leu Asp 385 390 395 400 Ser Asp Gly Ser Phe Phe Leu Val Ser Lys
Leu Thr Val Asp Lys Ser 405 410 415 Arg Trp Gln Gln Gly Asn Val Phe
Ser Cys Ser Val Leu His Glu Ala 420 425 430 Leu His Ser His Tyr Thr
Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys 435 440 445 <210> SEQ
ID NO 54 <211> LENGTH: 219 <212> TYPE: PRT <213>
ORGANISM: Artificial Sequence <220> FEATURE: <221>
NAME/KEY: source <223> OTHER INFORMATION: /note="Description
of Artificial Sequence: Synthetic polypeptide" <400>
SEQUENCE: 54 Asp Ile Val Met Thr Gln Thr Pro Leu Ser Leu Pro Val
Thr Pro Gly 1 5 10 15 Glu Pro Ala Ser Ile Ser Cys Gln Ser Ser Lys
Ser Leu Leu His Ser 20 25 30 Thr Gly Lys Thr Tyr Leu Asn Trp Tyr
Leu Gln Lys Pro Gly Gln Ser 35 40 45 Pro Gln Leu Leu Ile Tyr Trp
Met Ser Thr Arg Ala Ser Gly Val Pro 50 55 60 Asp Arg Phe Ser Gly
Ser Gly Ser Gly Thr Asp Phe Thr Leu Lys Ile 65 70 75 80 Ser Arg Val
Glu Ala Glu Asp Val Gly Val Tyr Tyr Cys Gln Gln Phe 85 90 95 Leu
Glu Tyr Pro Phe Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys 100 105
110 Arg Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu
115 120 125 Gln Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn
Asn Phe 130 135 140 Tyr Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp
Asn Ala Leu Gln 145 150 155 160 Ser Gly Asn Ser Gln Glu Ser Val Thr
Glu Gln Asp Ser Lys Asp Ser 165 170 175 Thr Tyr Ser Leu Ser Ser Thr
Leu Thr Leu Ser Lys Ala Asp Tyr Glu 180 185 190 Lys His Lys Val Tyr
Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser 195 200 205 Pro Val Thr
Lys Ser Phe Asn Arg Gly Glu Cys 210 215 <210> SEQ ID NO 55
<211> LENGTH: 181 <212> TYPE: PRT <213> ORGANISM:
Homo sapiens <400> SEQUENCE: 55 Asn Ser Val Ile Ile Val Asp
Lys Asn Gly Arg Leu Val Tyr Leu Val 1 5 10 15 Glu Asn Pro Gly Gly
Tyr Val Ala Tyr Ser Lys Ala Ala Thr Val Thr 20 25 30 Gly Lys Leu
Val His Ala Asn Phe Gly Thr Lys Lys Asp Phe Glu Asp 35 40 45 Leu
Tyr Thr Pro Val Asn Gly Ser Ile Val Ile Val Arg Ala Gly Lys 50 55
60 Ile Thr Phe Ala Glu Lys Val Ala Asn Ala Glu Ser Leu Asn Ala Ile
65 70 75 80 Gly Val Leu Ile Tyr Met Asp Gln Thr Lys Phe Pro Ile Val
Asn Ala 85 90 95 Glu Leu Ser Phe Phe Gly His Ala His Leu Gly Thr
Gly Asp Pro Tyr 100 105 110 Thr Pro Gly Phe Pro Ser Phe Asn His Thr
Gln Phe Pro Pro Ser Arg 115 120 125 Ser Ser Gly Leu Pro Asn Ile Pro
Val Gln Thr Ile Ser Arg Ala Ala 130 135 140 Ala Glu Lys Leu Phe Gly
Asn Met Glu Gly Asp Cys Pro Ser Asp Trp 145 150 155 160 Lys Thr Asp
Ser Thr Cys Arg Met Val Thr Ser Glu Ser Lys Asn Val 165 170 175 Lys
Leu Thr Val Ser 180 <210> SEQ ID NO 56 <211> LENGTH: 10
<212> TYPE: PRT <213> ORGANISM: Artificial Sequence
<220> FEATURE: <221> NAME/KEY: source <223> OTHER
INFORMATION: /note="Description of Artificial Sequence: Synthetic
peptide" <400> SEQUENCE: 56 Gly Gly Gly Gly Ser Gly Gly Gly
Gly Ser 1 5 10 <210> SEQ ID NO 57 <211> LENGTH: 10
<212> TYPE: PRT <213> ORGANISM: Homo sapiens
<400> SEQUENCE: 57 Asp Lys Thr His Thr Cys Pro Pro Cys Pro 1
5 10 <210> SEQ ID NO 58 <211> LENGTH: 7 <212>
TYPE: PRT <213> ORGANISM: Artificial Sequence <220>
FEATURE: <221> NAME/KEY: source <223> OTHER
INFORMATION: /note="Description of Artificial Sequence: Synthetic
peptide" <220> FEATURE: <221> NAME/KEY: MOD_RES
<222> LOCATION: (2)..(2) <223> OTHER INFORMATION: Any
amino acid <400> SEQUENCE: 58 Tyr Xaa Thr Glu Trp Ser Ser 1 5
<210> SEQ ID NO 59 <211> LENGTH: 9 <212> TYPE:
PRT <213> ORGANISM: Artificial Sequence <220> FEATURE:
<221> NAME/KEY: source <223> OTHER INFORMATION:
/note="Description of Artificial Sequence: Synthetic peptide"
<220> FEATURE: <221> NAME/KEY: MOD_RES <222>
LOCATION: (2)..(3) <223> OTHER INFORMATION: Any amino acid
<220> FEATURE: <221> NAME/KEY: MOD_RES <222>
LOCATION: (5)..(8) <223> OTHER INFORMATION: Any amino acid
<400> SEQUENCE: 59 Thr Xaa Xaa Glu Xaa Xaa Xaa Xaa Phe 1 5
<210> SEQ ID NO 60 <211> LENGTH: 760 <212> TYPE:
PRT <213> ORGANISM: Macaca fascicularis <400> SEQUENCE:
60 Met Met Asp Gln Ala Arg Ser Ala Phe Ser Asn Leu Phe Gly Gly Glu
1 5 10 15 Pro Leu Ser Tyr Thr Arg Phe Ser Leu Ala Arg Gln Val Asp
Gly Asp 20 25 30 Asn Ser His Val Glu Met Lys Leu Gly Val Asp Glu
Glu Glu Asn Thr 35 40 45 Asp Asn Asn Thr Lys Ala Asn Gly Thr Lys
Pro Lys Arg Cys Gly Gly 50 55 60 Asn Ile Cys Tyr Gly Thr Ile Ala
Val Ile Ile Phe Phe Leu Ile Gly 65 70 75 80 Phe Met Ile Gly Tyr Leu
Gly Tyr Cys Lys Gly Val Glu Pro Lys Thr 85 90 95 Glu Cys Glu Arg
Leu Ala Gly Thr Glu Ser Pro Ala Arg Glu Glu Pro 100 105 110 Glu Glu
Asp Phe Pro Ala Ala Pro Arg Leu Tyr Trp Asp Asp Leu Lys 115 120 125
Arg Lys Leu Ser Glu Lys Leu Asp Thr Thr Asp Phe Thr Ser Thr Ile 130
135 140 Lys Leu Leu Asn Glu Asn Leu Tyr Val Pro Arg Glu Ala Gly Ser
Gln 145 150 155 160 Lys Asp Glu Asn Leu Ala Leu Tyr Ile Glu Asn Gln
Phe Arg Glu Phe 165 170 175 Lys Leu Ser Lys Val Trp Arg Asp Gln His
Phe Val Lys Ile Gln Val 180 185 190 Lys Asp Ser Ala Gln Asn Ser Val
Ile Ile Val Asp Lys Asn Gly Gly 195 200 205 Leu Val Tyr Leu Val Glu
Asn Pro Gly Gly Tyr Val Ala Tyr Ser Lys 210 215 220 Ala Ala Thr Val
Thr Gly Lys Leu Val His Ala Asn Phe Gly Thr Lys 225 230 235 240 Lys
Asp Phe Glu Asp Leu Asp Ser Pro Val Asn Gly Ser Ile Val Ile 245 250
255 Val Arg Ala Gly Lys Ile Thr Phe Ala Glu Lys Val Ala Asn Ala Glu
260 265 270 Ser Leu Asn Ala Ile Gly Val Leu Ile Tyr Met Asp Gln Thr
Lys Phe 275 280 285 Pro Ile Val Lys Ala Asp Leu Ser Phe Phe Gly His
Ala His Leu Gly 290 295 300 Thr Gly Asp Pro Tyr Thr Pro Gly Phe Pro
Ser Phe Asn His Thr Gln 305 310 315 320 Phe Pro Pro Ser Gln Ser Ser
Gly Leu Pro Asn Ile Pro Val Gln Thr 325 330 335 Ile Ser Arg Ala Ala
Ala Glu Lys Leu Phe Gly Asn Met Glu Gly Asp 340 345 350 Cys Pro Ser
Asp Trp Lys Thr Asp Ser Thr Cys Lys Met Val Thr Ser 355 360 365 Glu
Asn Lys Ser Val Lys Leu Thr Val Ser Asn Val Leu Lys Glu Thr 370 375
380 Lys Ile Leu Asn Ile Phe Gly Val Ile Lys Gly Phe Val Glu Pro Asp
385 390 395 400 His Tyr Val Val Val Gly Ala Gln Arg Asp Ala Trp Gly
Pro Gly Ala 405 410 415 Ala Lys Ser Ser Val Gly Thr Ala Leu Leu Leu
Lys Leu Ala Gln Met 420 425 430 Phe Ser Asp Met Val Leu Lys Asp Gly
Phe Gln Pro Ser Arg Ser Ile 435 440 445 Ile Phe Ala Ser Trp Ser Ala
Gly Asp Phe Gly Ser Val Gly Ala Thr 450 455 460 Glu Trp Leu Glu Gly
Tyr Leu Ser Ser Leu His Leu Lys Ala Phe Thr 465 470 475 480 Tyr Ile
Asn Leu Asp Lys Ala Val Leu Gly Thr Ser Asn Phe Lys Val 485 490 495
Ser Ala Ser Pro Leu Leu Tyr Thr Leu Ile Glu Lys Thr Met Gln Asp 500
505 510 Val Lys His Pro Val Thr Gly Arg Ser Leu Tyr Gln Asp Ser Asn
Trp 515 520 525 Ala Ser Lys Val Glu Lys Leu Thr Leu Asp Asn Ala Ala
Phe Pro Phe 530 535 540 Leu Ala Tyr Ser Gly Ile Pro Ala Val Ser Phe
Cys Phe Cys Glu Asp 545 550 555 560 Thr Asp Tyr Pro Tyr Leu Gly Thr
Thr Met Asp Thr Tyr Lys Glu Leu 565 570 575 Val Glu Arg Ile Pro Glu
Leu Asn Lys Val Ala Arg Ala Ala Ala Glu 580 585 590 Val Ala Gly Gln
Phe Val Ile Lys Leu Thr His Asp Thr Glu Leu Asn 595 600 605 Leu Asp
Tyr Glu Arg Tyr Asn Ser Gln Leu Leu Leu Phe Leu Arg Asp 610 615 620
Leu Asn Gln Tyr Arg Ala Asp Val Lys Glu Met Gly Leu Ser Leu Gln 625
630 635 640 Trp Leu Tyr Ser Ala Arg Gly Asp Phe Phe Arg Ala Thr Ser
Arg Leu 645 650 655 Thr Thr Asp Phe Arg Asn Ala Glu Lys Arg Asp Lys
Phe Val Met Lys 660 665 670 Lys Leu Asn Asp Arg Val Met Arg Val Glu
Tyr Tyr Phe Leu Ser Pro 675 680 685 Tyr Val Ser Pro Lys Glu Ser Pro
Phe Arg His Val Phe Trp Gly Ser 690 695 700 Gly Ser His Thr Leu Ser
Ala Leu Leu Glu Ser Leu Lys Leu Arg Arg 705 710 715 720 Gln Asn Asn
Ser Ala Phe Asn Glu Thr Leu Phe Arg Asn Gln Leu Ala 725 730 735 Leu
Ala Thr Trp Thr Ile Gln Gly Ala Ala Asn Ala Leu Ser Gly Asp 740 745
750 Val Trp Asp Ile Asp Asn Glu Phe 755 760 <210> SEQ ID NO
61 <211> LENGTH: 217 <212> TYPE: PRT <213>
ORGANISM: Artificial Sequence <220> FEATURE: <221>
NAME/KEY: source <223> OTHER INFORMATION: /note="Description
of Artificial Sequence: Synthetic polypeptide" <400>
SEQUENCE: 61 Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe
Pro Pro Lys 1 5 10 15 Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro
Glu Val Thr Cys Val 20 25 30 Val Val Asp Val Ser His Glu Asp Pro
Glu Val Lys Phe Asn Trp Tyr 35 40 45 Val Asp Gly Val Glu Val His
Asn Ala Lys Thr Lys Pro Arg Glu Glu 50 55 60 Gln Tyr Asn Ser Thr
Tyr Arg Val Val Ser Val Leu Thr Val Leu His 65 70 75 80 Gln Asp Trp
Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys 85 90 95 Ala
Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln 100 105
110 Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu
115 120 125 Thr Lys Asn Gln Val Ser Leu Ser Cys Ala Val Lys Gly Phe
Tyr Pro 130 135 140 Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln
Pro Glu Asn Asn 145 150 155 160 Tyr Lys Thr Thr Pro Pro Val Leu Asp
Ser Asp Gly Ser Phe Phe Leu 165 170 175 Val Ser Lys Leu Thr Val Asp
Lys Ser Arg Trp Gln Gln Gly Asn Val 180 185 190 Phe Ser Cys Ser Val
Leu His Glu Ala Leu His Ser His Tyr Thr Gln 195 200 205 Lys Ser Leu
Ser Leu Ser Pro Gly Lys 210 215 <210> SEQ ID NO 62
<211> LENGTH: 760 <212> TYPE: PRT <213> ORGANISM:
Homo sapiens <400> SEQUENCE: 62 Met Met Asp Gln Ala Arg Ser
Ala Phe Ser Asn Leu Phe Gly Gly Glu 1 5 10 15 Pro Leu Ser Tyr Thr
Arg Phe Ser Leu Ala Arg Gln Val Asp Gly Asp 20 25 30 Asn Ser His
Val Glu Met Lys Leu Ala Val Asp Glu Glu Glu Asn Ala 35 40 45 Asp
Asn Asn Thr Lys Ala Asn Val Thr Lys Pro Lys Arg Cys Ser Gly 50 55
60 Ser Ile Cys Tyr Gly Thr Ile Ala Val Ile Val Phe Phe Leu Ile Gly
65 70 75 80 Phe Met Ile Gly Tyr Leu Gly Tyr Cys Lys Gly Val Glu Pro
Lys Thr 85 90 95 Glu Cys Glu Arg Leu Ala Gly Thr Glu Ser Pro Val
Arg Glu Glu Pro 100 105 110 Gly Glu Asp Phe Pro Ala Ala Arg Arg Leu
Tyr Trp Asp Asp Leu Lys 115 120 125 Arg Lys Leu Ser Glu Lys Leu Asp
Ser Thr Asp Phe Thr Gly Thr Ile 130 135 140 Lys Leu Leu Asn Glu Asn
Ser Tyr Val Pro Arg Glu Ala Gly Ser Gln 145 150 155 160 Lys Asp Glu
Asn Leu Ala Leu Tyr Val Glu Asn Gln Phe Arg Glu Phe 165 170 175 Lys
Leu Ser Lys Val Trp Arg Asp Gln His Phe Val Lys Ile Gln Val 180 185
190 Lys Asp Ser Ala Gln Asn Ser Val Ile Ile Val Asp Lys Asn Gly Arg
195 200 205 Leu Val Tyr Leu Val Glu Asn Pro Gly Gly Tyr Val Ala Tyr
Ser Lys 210 215 220 Ala Ala Thr Val Thr Gly Lys Leu Val His Ala Asn
Phe Gly Thr Lys 225 230 235 240 Lys Asp Phe Glu Asp Leu Tyr Thr Pro
Val Asn Gly Ser Ile Val Ile 245 250 255 Val Arg Ala Gly Lys Ile Thr
Phe Ala Glu Lys Val Ala Asn Ala Glu 260 265 270 Ser Leu Asn Ala Ile
Gly Val Leu Ile Tyr Met Asp Gln Thr Lys Phe 275 280 285 Pro Ile Val
Asn Ala Glu Leu Ser Phe Phe Gly His Ala His Leu Gly 290 295 300 Thr
Gly Asp Pro Tyr Thr Pro Gly Phe Pro Ser Phe Asn His Thr Gln 305 310
315 320 Phe Pro Pro Ser Arg Ser Ser Gly Leu Pro Asn Ile Pro Val Gln
Thr 325 330 335 Ile Ser Arg Ala Ala Ala Glu Lys Leu Phe Gly Asn Met
Glu Gly Asp 340 345 350 Cys Pro Ser Asp Trp Lys Thr Asp Ser Thr Cys
Arg Met Val Thr Ser 355 360 365 Glu Ser Lys Asn Val Lys Leu Thr Val
Ser Asn Val Leu Lys Glu Ile 370 375 380 Lys Ile Leu Asn Ile Phe Gly
Val Ile Lys Gly Phe Val Glu Pro Asp 385 390 395 400 His Tyr Val Val
Val Gly Ala Gln Arg Asp Ala Trp Gly Pro Gly Ala 405 410 415 Ala Lys
Ser Gly Val Gly Thr Ala Leu Leu Leu Lys Leu Ala Gln Met 420 425 430
Phe Ser Asp Met Val Leu Lys Asp Gly Phe Gln Pro Ser Arg Ser Ile 435
440 445 Ile Phe Ala Ser Trp Ser Ala Gly Asp Phe Gly Ser Val Gly Ala
Thr 450 455 460 Glu Trp Leu Glu Gly Tyr Leu Ser Ser Leu His Leu Lys
Ala Phe Thr 465 470 475 480 Tyr Ile Asn Leu Asp Lys Ala Val Leu Gly
Thr Ser Asn Phe Lys Val 485 490 495 Ser Ala Ser Pro Leu Leu Tyr Thr
Leu Ile Glu Lys Thr Met Gln Asn 500 505 510 Val Lys His Pro Val Thr
Gly Gln Phe Leu Tyr Gln Asp Ser Asn Trp 515 520 525 Ala Ser Lys Val
Glu Lys Leu Thr Leu Asp Asn Ala Ala Phe Pro Phe 530 535 540 Leu Ala
Tyr Ser Gly Ile Pro Ala Val Ser Phe Cys Phe Cys Glu Asp 545 550 555
560 Thr Asp Tyr Pro Tyr Leu Gly Thr Thr Met Asp Thr Tyr Lys Glu Leu
565 570 575 Ile Glu Arg Ile Pro Glu Leu Asn Lys Val Ala Arg Ala Ala
Ala Glu 580 585 590 Val Ala Gly Gln Phe Val Ile Lys Leu Thr His Asp
Val Glu Leu Asn 595 600 605 Leu Asp Tyr Glu Arg Tyr Asn Ser Gln Leu
Leu Ser Phe Val Arg Asp 610 615 620 Leu Asn Gln Tyr Arg Ala Asp Ile
Lys Glu Met Gly Leu Ser Leu Gln 625 630 635 640 Trp Leu Tyr Ser Ala
Arg Gly Asp Phe Phe Arg Ala Thr Ser Arg Leu 645 650 655 Thr Thr Asp
Phe Gly Asn Ala Glu Lys Thr Asp Arg Phe Val Met Lys 660 665 670 Lys
Leu Asn Asp Arg Val Met Arg Val Glu Tyr His Phe Leu Ser Pro 675 680
685 Tyr Val Ser Pro Lys Glu Ser Pro Phe Arg His Val Phe Trp Gly Ser
690 695 700 Gly Ser His Thr Leu Pro Ala Leu Leu Glu Asn Leu Lys Leu
Arg Lys 705 710 715 720 Gln Asn Asn Gly Ala Phe Asn Glu Thr Leu Phe
Arg Asn Gln Leu Ala 725 730 735 Leu Ala Thr Trp Thr Ile Gln Gly Ala
Ala Asn Ala Leu Ser Gly Asp 740 745 750 Val Trp Asp Ile Asp Asn Glu
Phe 755 760 <210> SEQ ID NO 63 <211> LENGTH: 216
<212> TYPE: PRT <213> ORGANISM: Artificial Sequence
<220> FEATURE: <221> NAME/KEY: source <223> OTHER
INFORMATION: /note="Description of Artificial Sequence: Synthetic
polypeptide" <400> SEQUENCE: 63 Ala Pro Glu Ala Ala Gly Gly
Pro Ser Val Phe Leu Phe Pro Pro Lys 1 5 10 15 Pro Lys Asp Thr Leu
Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val 20 25 30 Val Val Asp
Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr 35 40 45 Val
Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu 50 55
60 Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His
65 70 75 80 Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser
Asn Lys 85 90 95 Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys
Ala Lys Gly Gln 100 105 110 Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro
Pro Ser Arg Asp Glu Leu 115 120 125 Thr Lys Asn Gln Val Ser Leu Ser
Cys Ala Val Lys Gly Phe Tyr Pro 130 135 140 Ser Asp Ile Ala Val Glu
Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn 145 150 155 160 Tyr Lys Thr
Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu 165 170 175 Val
Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val 180 185
190 Phe Ser Cys Ser Val Leu His Glu Ala Leu His Ser His Tyr Thr Gln
195 200 205 Lys Ser Leu Ser Leu Ser Pro Gly 210 215 <210> SEQ
ID NO 64 <211> LENGTH: 216 <212> TYPE: PRT <213>
ORGANISM: Artificial Sequence <220> FEATURE: <221>
NAME/KEY: source <223> OTHER INFORMATION: /note="Description
of Artificial Sequence: Synthetic polypeptide" <400>
SEQUENCE: 64 Ala Pro Glu Ala Ala Gly Gly Pro Ser Val Phe Leu Phe
Pro Pro Lys 1 5 10 15 Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro
Glu Val Thr Cys Val 20 25 30 Val Val Asp Val Ser His Glu Asp Pro
Glu Val Lys Phe Asn Trp Tyr 35 40 45 Val Asp Gly Val Glu Val His
Asn Ala Lys Thr Lys Pro Arg Glu Glu 50 55 60 Gln Tyr Asn Ser Thr
Tyr Arg Val Val Ser Val Leu Thr Val Leu His 65 70 75 80 Gln Asp Trp
Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys 85 90 95 Ala
Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln 100 105
110 Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu
115 120 125 Thr Lys Asn Gln Val Ser Leu Trp Cys Leu Val Lys Gly Phe
Tyr Pro 130 135 140 Ser Asp Ile Ala Val Trp Trp Glu Ser Tyr Gly Thr
Glu Trp Ser Ser 145 150 155 160 Tyr Lys Thr Thr Pro Pro Val Leu Asp
Ser Asp Gly Ser Phe Phe Leu 165 170 175 Tyr Ser Lys Leu Thr Val Thr
Lys Glu Glu Trp Gln Gln Gly Phe Val 180 185 190 Phe Ser Cys Ser Val
Leu His Glu Ala Leu His Ser His Tyr Thr Gln 195 200 205 Lys Ser Leu
Ser Leu Ser Pro Gly 210 215 <210> SEQ ID NO 65 <211>
LENGTH: 447 <212> TYPE: PRT <213> ORGANISM: Artificial
Sequence <220> FEATURE: <221> NAME/KEY: source
<223> OTHER INFORMATION: /note="Description of Artificial
Sequence: Synthetic polypeptide" <400> SEQUENCE: 65 Glu Val
Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Thr Asp Phe 20
25 30 Tyr Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp
Val 35 40 45 Ser Val Ile Arg Asn Lys Ala Asn Ala Tyr Thr Ala Gly
Tyr Asn Pro 50 55 60 Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp
Asn Ser Lys Asn Thr 65 70 75 80 Leu Tyr Leu Gln Met Asn Ser Leu Arg
Ala Glu Asp Thr Ala Val Tyr 85 90 95 Tyr Cys Ala Arg Leu Thr Tyr
Gly Phe Asp Tyr Trp Gly Gln Gly Thr 100 105 110 Leu Val Thr Val Ser
Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro 115 120 125 Leu Ala Pro
Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly 130 135 140 Cys
Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn 145 150
155 160 Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu
Gln 165 170 175 Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val
Pro Ser Ser 180 185 190 Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val
Asn His Lys Pro Ser 195 200 205 Asn Thr Lys Val Asp Lys Lys Val Glu
Pro Lys Ser Cys Asp Lys Thr 210 215 220 His Thr Cys Pro Pro Cys Pro
Ala Pro Glu Ala Ala Gly Gly Pro Ser 225 230 235 240 Val Phe Leu Phe
Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg 245 250 255 Thr Pro
Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro 260 265 270
Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala 275
280 285 Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val
Val 290 295 300 Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly
Lys Glu Tyr 305 310 315 320 Lys Cys Lys Val Ser Asn Lys Ala Leu Pro
Ala Pro Ile Glu Lys Thr 325 330 335 Ile Ser Lys Ala Lys Gly Gln Pro
Arg Glu Pro Gln Val Tyr Thr Leu 340 345 350 Pro Pro Ser Arg Asp Glu
Leu Thr Lys Asn Gln Val Ser Leu Trp Cys 355 360 365 Leu Val Lys Gly
Phe Tyr Pro Ser Asp Ile Ala Val Trp Trp Glu Ser 370 375 380 Tyr Gly
Thr Glu Trp Ser Ser Tyr Lys Thr Thr Pro Pro Val Leu Asp 385 390 395
400 Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Thr Lys Glu
405 410 415 Glu Trp Gln Gln Gly Phe Val Phe Ser Cys Ser Val Leu His
Glu Ala 420 425 430 Leu His Ser His Tyr Thr Gln Lys Ser Leu Ser Leu
Ser Pro Gly 435 440 445 <210> SEQ ID NO 66 <211>
LENGTH: 216 <212> TYPE: PRT <213> ORGANISM: Artificial
Sequence <220> FEATURE: <221> NAME/KEY: source
<223> OTHER INFORMATION: /note="Description of Artificial
Sequence: Synthetic polypeptide" <400> SEQUENCE: 66 Ala Pro
Glu Ala Ala Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys 1 5 10 15
Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val 20
25 30 Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp
Tyr 35 40 45 Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro
Arg Glu Glu 50 55 60 Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val
Leu Thr Val Leu His 65 70 75 80 Gln Asp Trp Leu Asn Gly Lys Glu Tyr
Lys Cys Lys Val Ser Asn Lys 85 90 95 Ala Leu Pro Ala Pro Ile Glu
Lys Thr Ile Ser Lys Ala Lys Gly Gln 100 105 110 Pro Arg Glu Pro Gln
Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu 115 120 125 Thr Lys Asn
Gln Val Ser Leu Trp Cys Leu Val Lys Gly Phe Tyr Pro 130 135 140 Ser
Asp Ile Ala Val Glu Trp Glu Ser Phe Gly Thr Glu Trp Ser Asn 145 150
155 160 Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe
Leu 165 170 175 Tyr Ser Lys Leu Thr Val Ser Lys Glu Glu Trp Gln Gln
Gly Phe Val 180 185 190 Phe Ser Cys Ser Val Leu His Glu Ala Leu His
Ser His Tyr Thr Gln 195 200 205 Lys Ser Leu Ser Leu Ser Pro Gly 210
215 <210> SEQ ID NO 67 <211> LENGTH: 447 <212>
TYPE: PRT <213> ORGANISM: Artificial Sequence <220>
FEATURE: <221> NAME/KEY: source <223> OTHER
INFORMATION: /note="Description of Artificial Sequence: Synthetic
polypeptide" <400> SEQUENCE: 67 Glu Val Gln Leu Val Glu Ser
Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser
Cys Ala Ala Ser Gly Phe Thr Phe Thr Asp Phe 20 25 30 Tyr Met Ser
Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser
Val Ile Arg Asn Lys Ala Asn Ala Tyr Thr Ala Gly Tyr Asn Pro 50 55
60 Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr
65 70 75 80 Leu Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala
Val Tyr 85 90 95 Tyr Cys Ala Arg Leu Thr Tyr Gly Phe Asp Tyr Trp
Gly Gln Gly Thr 100 105 110 Leu Val Thr Val Ser Ser Ala Ser Thr Lys
Gly Pro Ser Val Phe Pro 115 120 125 Leu Ala Pro Ser Ser Lys Ser Thr
Ser Gly Gly Thr Ala Ala Leu Gly 130 135 140 Cys Leu Val Lys Asp Tyr
Phe Pro Glu Pro Val Thr Val Ser Trp Asn 145 150 155 160 Ser Gly Ala
Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln 165 170 175 Ser
Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser 180 185
190 Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser
195 200 205 Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp
Lys Thr 210 215 220 His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala
Gly Gly Pro Ser 225 230 235 240 Val Phe Leu Phe Pro Pro Lys Pro Lys
Asp Thr Leu Met Ile Ser Arg 245 250 255 Thr Pro Glu Val Thr Cys Val
Val Val Asp Val Ser His Glu Asp Pro 260 265 270 Glu Val Lys Phe Asn
Trp Tyr Val Asp Gly Val Glu Val His Asn Ala 275 280 285 Lys Thr Lys
Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val 290 295 300 Ser
Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr 305 310
315 320 Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys
Thr 325 330 335 Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val
Tyr Thr Leu 340 345 350 Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln
Val Ser Leu Trp Cys 355 360 365 Leu Val Lys Gly Phe Tyr Pro Ser Asp
Ile Ala Val Glu Trp Glu Ser 370 375 380 Phe Gly Thr Glu Trp Ser Asn
Tyr Lys Thr Thr Pro Pro Val Leu Asp 385 390 395 400 Ser Asp Gly Ser
Phe Phe Leu Tyr Ser Lys Leu Thr Val Ser Lys Glu 405 410 415 Glu Trp
Gln Gln Gly Phe Val Phe Ser Cys Ser Val Leu His Glu Ala 420 425 430
Leu His Ser His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly 435 440
445 <210> SEQ ID NO 68 <211> LENGTH: 216 <212>
TYPE: PRT <213> ORGANISM: Artificial Sequence <220>
FEATURE: <221> NAME/KEY: source <223> OTHER
INFORMATION: /note="Description of Artificial Sequence: Synthetic
polypeptide" <400> SEQUENCE: 68 Ala Pro Glu Ala Ala Gly Gly
Pro Ser Val Phe Leu Phe Pro Pro Lys 1 5 10 15 Pro Lys Asp Thr Leu
Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val 20 25 30 Val Val Asp
Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr 35 40 45 Val
Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu 50 55
60 Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His
65 70 75 80 Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser
Asn Lys 85 90 95 Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys
Ala Lys Gly Gln 100 105 110 Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro
Pro Ser Arg Asp Glu Leu 115 120 125 Thr Lys Asn Gln Val Ser Leu Trp
Cys Leu Val Lys Gly Phe Tyr Pro 130 135 140 Ser Asp Ile Ala Val Glu
Trp Glu Ser Tyr Gly Thr Glu Trp Val Asn 145 150 155 160 Tyr Lys Thr
Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu 165 170 175 Tyr
Ser Lys Leu Thr Val Thr Lys Glu Glu Trp Gln Gln Gly Phe Val 180 185
190 Phe Ser Cys Ser Val Leu His Glu Ala Leu His Ser His Tyr Thr Gln
195 200 205 Lys Ser Leu Ser Leu Ser Pro Gly 210 215 <210> SEQ
ID NO 69 <211> LENGTH: 447 <212> TYPE: PRT <213>
ORGANISM: Artificial Sequence <220> FEATURE: <221>
NAME/KEY: source <223> OTHER INFORMATION: /note="Description
of Artificial Sequence: Synthetic polypeptide" <400>
SEQUENCE: 69 Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln
Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe
Thr Phe Thr Asp Phe 20 25 30 Tyr Met Ser Trp Val Arg Gln Ala Pro
Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Val Ile Arg Asn Lys Ala
Asn Ala Tyr Thr Ala Gly Tyr Asn Pro 50 55 60 Ser Val Lys Gly Arg
Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr 65 70 75 80 Leu Tyr Leu
Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr 85 90 95 Tyr
Cys Ala Arg Leu Thr Tyr Gly Phe Asp Tyr Trp Gly Gln Gly Thr 100 105
110 Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro
115 120 125 Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala
Leu Gly 130 135 140 Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr
Val Ser Trp Asn 145 150 155 160 Ser Gly Ala Leu Thr Ser Gly Val His
Thr Phe Pro Ala Val Leu Gln 165 170 175 Ser Ser Gly Leu Tyr Ser Leu
Ser Ser Val Val Thr Val Pro Ser Ser 180 185 190 Ser Leu Gly Thr Gln
Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser 195 200 205 Asn Thr Lys
Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr 210 215 220 His
Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Gly Pro Ser 225 230
235 240 Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser
Arg 245 250 255 Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His
Glu Asp Pro 260 265 270 Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val
Glu Val His Asn Ala 275 280 285 Lys Thr Lys Pro Arg Glu Glu Gln Tyr
Asn Ser Thr Tyr Arg Val Val 290 295 300 Ser Val Leu Thr Val Leu His
Gln Asp Trp Leu Asn Gly Lys Glu Tyr 305 310 315 320 Lys Cys Lys Val
Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr 325 330 335 Ile Ser
Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu 340 345 350
Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Trp Cys 355
360 365 Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu
Ser 370 375 380 Tyr Gly Thr Glu Trp Val Asn Tyr Lys Thr Thr Pro Pro
Val Leu Asp 385 390 395 400 Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys
Leu Thr Val Thr Lys Glu 405 410 415 Glu Trp Gln Gln Gly Phe Val Phe
Ser Cys Ser Val Leu His Glu Ala 420 425 430 Leu His Ser His Tyr Thr
Gln Lys Ser Leu Ser Leu Ser Pro Gly 435 440 445 <210> SEQ ID
NO 70 <211> LENGTH: 216 <212> TYPE: PRT <213>
ORGANISM: Artificial Sequence <220> FEATURE: <221>
NAME/KEY: source <223> OTHER INFORMATION: /note="Description
of Artificial Sequence: Synthetic polypeptide" <400>
SEQUENCE: 70 Ala Pro Glu Ala Ala Gly Gly Pro Ser Val Phe Leu Phe
Pro Pro Lys 1 5 10 15 Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro
Glu Val Thr Cys Val 20 25 30 Val Val Asp Val Ser His Glu Asp Pro
Glu Val Lys Phe Asn Trp Tyr 35 40 45 Val Asp Gly Val Glu Val His
Asn Ala Lys Thr Lys Pro Arg Glu Glu 50 55 60 Gln Tyr Asn Ser Thr
Tyr Arg Val Val Ser Val Leu Thr Val Leu His 65 70 75 80 Gln Asp Trp
Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys 85 90 95 Ala
Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln 100 105
110 Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu
115 120 125 Thr Lys Asn Gln Val Ser Leu Trp Cys Leu Val Lys Gly Phe
Tyr Pro 130 135 140 Ser Asp Ile Ala Val Glu Trp Glu Ser Tyr Gly Thr
Glu Trp Ser Asn 145 150 155 160 Tyr Lys Thr Thr Pro Pro Val Leu Asp
Ser Asp Gly Ser Phe Phe Leu 165 170 175 Tyr Ser Lys Leu Thr Val Ser
Lys Glu Glu Trp Gln Gln Gly Phe Val 180 185 190 Phe Ser Cys Ser Val
Leu His Glu Ala Leu His Ser His Tyr Thr Gln 195 200 205 Lys Ser Leu
Ser Leu Ser Pro Gly 210 215 <210> SEQ ID NO 71 <211>
LENGTH: 447 <212> TYPE: PRT <213> ORGANISM: Artificial
Sequence <220> FEATURE: <221> NAME/KEY: source
<223> OTHER INFORMATION: /note="Description of Artificial
Sequence: Synthetic polypeptide" <400> SEQUENCE: 71 Glu Val
Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Thr Asp Phe 20
25 30 Tyr Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp
Val 35 40 45 Ser Val Ile Arg Asn Lys Ala Asn Ala Tyr Thr Ala Gly
Tyr Asn Pro 50 55 60 Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp
Asn Ser Lys Asn Thr 65 70 75 80 Leu Tyr Leu Gln Met Asn Ser Leu Arg
Ala Glu Asp Thr Ala Val Tyr 85 90 95 Tyr Cys Ala Arg Leu Thr Tyr
Gly Phe Asp Tyr Trp Gly Gln Gly Thr 100 105 110 Leu Val Thr Val Ser
Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro 115 120 125 Leu Ala Pro
Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly 130 135 140 Cys
Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn 145 150
155 160 Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu
Gln 165 170 175 Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val
Pro Ser Ser 180 185 190 Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val
Asn His Lys Pro Ser 195 200 205 Asn Thr Lys Val Asp Lys Lys Val Glu
Pro Lys Ser Cys Asp Lys Thr 210 215 220 His Thr Cys Pro Pro Cys Pro
Ala Pro Glu Ala Ala Gly Gly Pro Ser 225 230 235 240 Val Phe Leu Phe
Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg 245 250 255 Thr Pro
Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro 260 265 270
Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala 275
280 285 Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val
Val 290 295 300 Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly
Lys Glu Tyr 305 310 315 320 Lys Cys Lys Val Ser Asn Lys Ala Leu Pro
Ala Pro Ile Glu Lys Thr 325 330 335 Ile Ser Lys Ala Lys Gly Gln Pro
Arg Glu Pro Gln Val Tyr Thr Leu 340 345 350 Pro Pro Ser Arg Asp Glu
Leu Thr Lys Asn Gln Val Ser Leu Trp Cys 355 360 365 Leu Val Lys Gly
Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser 370 375 380 Tyr Gly
Thr Glu Trp Ser Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp 385 390 395
400 Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Ser Lys Glu
405 410 415 Glu Trp Gln Gln Gly Phe Val Phe Ser Cys Ser Val Leu His
Glu Ala 420 425 430 Leu His Ser His Tyr Thr Gln Lys Ser Leu Ser Leu
Ser Pro Gly 435 440 445 <210> SEQ ID NO 72 <211>
LENGTH: 447 <212> TYPE: PRT <213> ORGANISM: Artificial
Sequence <220> FEATURE: <221> NAME/KEY: source
<223> OTHER INFORMATION: /note="Description of Artificial
Sequence: Synthetic polypeptide" <400> SEQUENCE: 72 Glu Val
Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Thr Asp Phe 20
25 30 Tyr Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp
Val 35 40 45 Ser Val Ile Arg Asn Lys Ala Asn Ala Tyr Thr Ala Gly
Tyr Asn Pro 50 55 60 Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp
Asn Ser Lys Asn Thr 65 70 75 80 Leu Tyr Leu Gln Met Asn Ser Leu Arg
Ala Glu Asp Thr Ala Val Tyr 85 90 95 Tyr Cys Ala Arg Leu Thr Tyr
Gly Phe Asp Tyr Trp Gly Gln Gly Thr 100 105 110 Leu Val Thr Val Ser
Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro 115 120 125 Leu Ala Pro
Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly 130 135 140 Cys
Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn 145 150
155 160 Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu
Gln 165 170 175 Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val
Pro Ser Ser 180 185 190 Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val
Asn His Lys Pro Ser 195 200 205 Asn Thr Lys Val Asp Lys Lys Val Glu
Pro Lys Ser Cys Asp Lys Thr 210 215 220 His Thr Cys Pro Pro Cys Pro
Ala Pro Glu Leu Leu Gly Gly Pro Ser 225 230 235 240 Val Phe Leu Phe
Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg 245 250 255 Thr Pro
Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro 260 265 270
Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala 275
280 285 Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val
Val 290 295 300 Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly
Lys Glu Tyr 305 310 315 320 Lys Cys Lys Val Ser Asn Lys Ala Leu Pro
Ala Pro Ile Glu Lys Thr 325 330 335 Ile Ser Lys Ala Lys Gly Gln Pro
Arg Glu Pro Gln Val Tyr Thr Leu 340 345 350 Pro Pro Ser Arg Asp Glu
Leu Thr Lys Asn Gln Val Ser Leu Ser Cys 355 360 365 Ala Val Lys Gly
Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser 370 375 380 Asn Gly
Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp 385 390 395
400 Ser Asp Gly Ser Phe Phe Leu Val Ser Lys Leu Thr Val Asp Lys Ser
405 410 415 Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Leu His
Glu Ala 420 425 430 Leu His Ser His Tyr Thr Gln Lys Ser Leu Ser Leu
Ser Pro Gly 435 440 445 <210> SEQ ID NO 73 <211>
LENGTH: 447 <212> TYPE: PRT <213> ORGANISM: Artificial
Sequence <220> FEATURE: <221> NAME/KEY: source
<223> OTHER INFORMATION: /note="Description of Artificial
Sequence: Synthetic polypeptide" <400> SEQUENCE: 73 Glu Val
Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Thr Asp Phe 20
25 30 Tyr Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp
Val 35 40 45 Ser Val Ile Arg Asn Lys Ala Asn Ala Tyr Thr Ala Gly
Tyr Asn Pro 50 55 60 Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp
Asn Ser Lys Asn Thr 65 70 75 80 Leu Tyr Leu Gln Met Asn Ser Leu Arg
Ala Glu Asp Thr Ala Val Tyr 85 90 95 Tyr Cys Ala Arg Leu Thr Tyr
Gly Phe Asp Tyr Trp Gly Gln Gly Thr 100 105 110 Leu Val Thr Val Ser
Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro 115 120 125 Leu Ala Pro
Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly 130 135 140 Cys
Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn 145 150
155 160 Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu
Gln 165 170 175 Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val
Pro Ser Ser 180 185 190 Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val
Asn His Lys Pro Ser 195 200 205 Asn Thr Lys Val Asp Lys Lys Val Glu
Pro Lys Ser Cys Asp Lys Thr 210 215 220 His Thr Cys Pro Pro Cys Pro
Ala Pro Glu Ala Ala Gly Gly Pro Ser 225 230 235 240 Val Phe Leu Phe
Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg 245 250 255 Thr Pro
Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro 260 265 270
Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala 275
280 285 Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val
Val 290 295 300 Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly
Lys Glu Tyr 305 310 315 320 Lys Cys Lys Val Ser Asn Lys Ala Leu Pro
Ala Pro Ile Glu Lys Thr 325 330 335 Ile Ser Lys Ala Lys Gly Gln Pro
Arg Glu Pro Gln Val Tyr Thr Leu 340 345 350 Pro Pro Ser Arg Asp Glu
Leu Thr Lys Asn Gln Val Ser Leu Ser Cys 355 360 365 Ala Val Lys Gly
Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser 370 375 380 Asn Gly
Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp 385 390 395
400 Ser Asp Gly Ser Phe Phe Leu Val Ser Lys Leu Thr Val Asp Lys Ser
405 410 415 Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Leu His
Glu Ala 420 425 430 Leu His Ser His Tyr Thr Gln Lys Ser Leu Ser Leu
Ser Pro Gly 435 440 445 <210> SEQ ID NO 74 <211>
LENGTH: 453 <212> TYPE: PRT <213> ORGANISM: Artificial
Sequence <220> FEATURE: <221> NAME/KEY: source
<223> OTHER INFORMATION: /note="Description of Artificial
Sequence: Synthetic polypeptide" <400> SEQUENCE: 74 Gln Val
Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala 1 5 10 15
Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Ala Phe Ser Ser Gln 20
25 30 Trp Met Asn Trp Val Arg Gln Ala Pro Gly Gln Arg Leu Glu Trp
Ile 35 40 45 Gly Arg Ile Tyr Pro Gly Gly Gly Asp Thr Asn Tyr Ala
Gly Lys Phe 50 55 60 Gln Gly Arg Val Thr Ile Thr Ala Asp Thr Ser
Ala Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu
Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Leu Leu Arg Asn Gln
Pro Gly Glu Ser Tyr Ala Met Asp Tyr 100 105 110 Trp Gly Gln Gly Thr
Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly 115 120 125 Pro Ser Val
Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly 130 135 140 Thr
Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val 145 150
155 160 Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr
Phe 165 170 175 Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser
Ser Val Val 180 185 190 Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr
Tyr Ile Cys Asn Val 195 200 205 Asn His Lys Pro Ser Asn Thr Lys Val
Asp Lys Lys Val Glu Pro Lys 210 215 220 Ser Cys Asp Lys Thr His Thr
Cys Pro Pro Cys Pro Ala Pro Glu Leu 225 230 235 240 Leu Gly Gly Pro
Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr 245 250 255 Leu Met
Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val 260 265 270
Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val 275
280 285 Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn
Ser 290 295 300 Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln
Asp Trp Leu 305 310 315 320 Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser
Asn Lys Ala Leu Pro Ala 325 330 335 Pro Ile Glu Lys Thr Ile Ser Lys
Ala Lys Gly Gln Pro Arg Glu Pro 340 345 350 Gln Val Tyr Thr Leu Pro
Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln 355 360 365 Val Ser Leu Thr
Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala 370 375 380 Val Glu
Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr 385 390 395
400 Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu
405 410 415 Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser
Cys Ser 420 425 430 Val Met His Glu Ala Leu His Asn His Tyr Thr Gln
Lys Ser Leu Ser 435 440 445 Leu Ser Pro Gly Lys 450 <210> SEQ
ID NO 75 <211> LENGTH: 219 <212> TYPE: PRT <213>
ORGANISM: Artificial Sequence <220> FEATURE: <221>
NAME/KEY: source <223> OTHER INFORMATION: /note="Description
of Artificial Sequence: Synthetic polypeptide" <400>
SEQUENCE: 75 Asp Val Val Met Thr Gln Ser Pro Asp Ser Leu Ala Val
Ser Leu Gly 1 5 10 15 Glu Arg Ala Thr Ile Asn Cys Arg Ser Ser Gln
Ser Leu Val His Ser 20 25 30 Asn Arg Tyr Thr Tyr Leu His Trp Tyr
Gln Gln Lys Pro Gly Gln Ser 35 40 45 Pro Lys Leu Leu Ile Tyr Lys
Val Ser Asn Arg Phe Ser Gly Val Pro 50 55 60 Asp Arg Phe Ser Gly
Ser Gly Ser Gly Thr Asp Phe Thr Leu Lys Ile 65 70 75 80 Ser Arg Val
Glu Ala Glu Asp Val Gly Val Tyr Tyr Cys Ser Gln Ser 85 90 95 Thr
Arg Val Pro Tyr Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys 100 105
110 Arg Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu
115 120 125 Gln Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn
Asn Phe 130 135 140 Tyr Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp
Asn Ala Leu Gln 145 150 155 160 Ser Gly Asn Ser Gln Glu Ser Val Thr
Glu Gln Asp Ser Lys Asp Ser 165 170 175 Thr Tyr Ser Leu Ser Ser Thr
Leu Thr Leu Ser Lys Ala Asp Tyr Glu 180 185 190 Lys His Lys Val Tyr
Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser 195 200 205 Pro Val Thr
Lys Ser Phe Asn Arg Gly Glu Cys 210 215 <210> SEQ ID NO 76
<211> LENGTH: 453 <212> TYPE: PRT <213> ORGANISM:
Artificial Sequence <220> FEATURE: <221> NAME/KEY:
source <223> OTHER INFORMATION: /note="Description of
Artificial Sequence: Synthetic polypeptide" <400> SEQUENCE:
76 Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala
1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Ala Phe Ser
Ser Gln 20 25 30 Trp Met Asn Trp Val Arg Gln Ala Pro Gly Gln Arg
Leu Glu Trp Ile 35 40 45 Gly Arg Ile Tyr Pro Gly Gly Gly Asp Thr
Asn Tyr Ala Gly Lys Phe 50 55 60 Gln Gly Arg Val Thr Ile Thr Ala
Asp Thr Ser Ala Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu
Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Leu Leu
Arg Asn Gln Pro Gly Glu Ser Tyr Ala Met Asp Tyr 100 105 110 Trp Gly
Gln Gly Thr Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly 115 120 125
Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly 130
135 140 Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro
Val 145 150 155 160 Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly
Val His Thr Phe 165 170 175 Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr
Ser Leu Ser Ser Val Val 180 185 190 Thr Val Pro Ser Ser Ser Leu Gly
Thr Gln Thr Tyr Ile Cys Asn Val 195 200 205 Asn His Lys Pro Ser Asn
Thr Lys Val Asp Lys Lys Val Glu Pro Lys 210 215 220 Ser Cys Asp Lys
Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu 225 230 235 240 Leu
Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr 245 250
255 Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val
260 265 270 Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp
Gly Val 275 280 285 Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu
Gln Tyr Asn Ser 290 295 300 Thr Tyr Arg Val Val Ser Val Leu Thr Val
Leu His Gln Asp Trp Leu 305 310 315 320 Asn Gly Lys Glu Tyr Lys Cys
Lys Val Ser Asn Lys Ala Leu Pro Ala 325 330 335 Ser Ile Glu Lys Thr
Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro 340 345 350 Gln Val Tyr
Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln 355 360 365 Val
Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala 370 375
380 Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr
385 390 395 400 Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr
Ser Lys Leu 405 410 415 Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn
Val Phe Ser Cys Ser 420 425 430 Val Met His Gly Ala Leu His Asn His
Tyr Thr Gln Lys Ser Leu Ser 435 440 445 Leu Ser Pro Gly Lys 450
<210> SEQ ID NO 77 <211> LENGTH: 445 <212> TYPE:
PRT <213> ORGANISM: Artificial Sequence <220> FEATURE:
<221> NAME/KEY: source <223> OTHER INFORMATION:
/note="Description of Artificial Sequence: Synthetic polypeptide"
<400> SEQUENCE: 77 Gln Val Gln Leu Gln Glu Ser Gly Pro Gly
Leu Val Lys Pro Ser Glu 1 5 10 15 Thr Leu Ser Leu Thr Cys Ala Val
Ser Gly Tyr Ser Ile Thr Ser Asp 20 25 30 Tyr Ala Trp Asn Trp Ile
Arg Gln Pro Pro Gly Lys Gly Leu Glu Trp 35 40 45 Ile Gly Tyr Met
Ser Tyr Ser Gly Ser Thr Arg Tyr Asn Pro Ser Leu 50 55 60 Arg Ser
Arg Val Thr Ile Ser Val Asp Thr Ser Lys Asn Gln Phe Ser 65 70 75 80
Leu Lys Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr Cys 85
90 95 Ala Arg Gly Trp Pro Leu Ala Tyr Trp Gly Gln Gly Thr Leu Val
Thr 100 105 110 Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro
Leu Ala Pro 115 120 125 Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala
Leu Gly Cys Leu Val 130 135 140 Lys Asp Tyr Phe Pro Glu Pro Val Thr
Val Ser Trp Asn Ser Gly Ala 145 150 155 160 Leu Thr Ser Gly Val His
Thr Phe Pro Ala Val Leu Gln Ser Ser Gly 165 170 175 Leu Tyr Ser Leu
Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly 180 185 190 Thr Gln
Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys 195 200 205
Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys 210
215 220 Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe
Leu 225 230 235 240 Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser
Arg Thr Pro Glu 245 250 255 Val Thr Cys Val Val Val Asp Val Ser His
Glu Asp Pro Glu Val Lys 260 265 270 Phe Asn Trp Tyr Val Asp Gly Val
Glu Val His Asn Ala Lys Thr Lys 275 280 285 Pro Arg Glu Glu Gln Tyr
Asn Ser Thr Tyr Arg Val Val Ser Val Leu 290 295 300 Thr Val Leu His
Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys 305 310 315 320 Val
Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys 325 330
335 Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser
340 345 350 Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Thr Cys Leu
Val Lys 355 360 365 Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu
Ser Asn Gly Gln 370 375 380 Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro
Val Leu Asp Ser Asp Gly 385 390 395 400 Ser Phe Phe Leu Tyr Ser Lys
Leu Thr Val Asp Lys Ser Arg Trp Gln 405 410 415 Gln Gly Asn Val Phe
Ser Cys Ser Val Met His Glu Ala Leu His Asn 420 425 430 His Tyr Thr
Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys 435 440 445 <210> SEQ
ID NO 78 <211> LENGTH: 214 <212> TYPE: PRT <213>
ORGANISM: Artificial Sequence <220> FEATURE: <221>
NAME/KEY: source <223> OTHER INFORMATION: /note="Description
of Artificial Sequence: Synthetic polypeptide" <400>
SEQUENCE: 78 Glu Ile Val Met Thr Gln Ser Pro Ala Thr Leu Ser Leu
Ser Pro Gly 1 5 10 15 Glu Arg Ala Thr Leu Ser Cys Ser Ala Ser Ser
Ser Val Ser Tyr Met 20 25 30 Tyr Trp Tyr Gln Gln Lys Pro Gly Gln
Ala Pro Arg Leu Leu Ile Tyr 35 40 45 Asp Thr Ser Asn Leu Ala Ser
Gly Ile Pro Ala Arg Phe Ser Gly Ser 50 55 60 Gly Ser Gly Thr Asp
Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro Glu 65 70 75 80 Asp Phe Ala
Val Tyr Tyr Cys Gln Gln Trp Ser Ser Tyr Pro Pro Ile 85 90 95 Thr
Phe Gly Gln Gly Thr Lys Val Glu Ile Lys Arg Thr Val Ala Ala 100 105
110 Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly
115 120 125 Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg
Glu Ala 130 135 140 Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser
Gly Asn Ser Gln 145 150 155 160 Glu Ser Val Thr Glu Gln Asp Ser Lys
Asp Ser Thr Tyr Ser Leu Ser 165 170 175 Ser Thr Leu Thr Leu Ser Lys
Ala Asp Tyr Glu Lys His Lys Val Tyr 180 185 190 Ala Cys Glu Val Thr
His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser 195 200 205 Phe Asn Arg
Gly Glu Cys 210 <210> SEQ ID NO 79 <211> LENGTH: 450
<212> TYPE: PRT <213> ORGANISM: Artificial Sequence
<220> FEATURE: <221> NAME/KEY: source <223> OTHER
INFORMATION: /note="Description of Artificial Sequence: Synthetic
polypeptide" <400> SEQUENCE: 79 Gln Val Thr Leu Arg 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 Thr Ser 20 25 30 Gly Met Ser
Val Gly Trp Ile Arg Gln Pro Pro Gly Lys Ala Leu Glu 35 40 45 Trp
Leu Ala Asp Ile Trp Trp Asp Asp Lys Lys Asp Tyr Asn Pro Ser 50 55
60 Leu Lys Ser Arg Leu Thr Ile Ser Lys Asp Thr Ser Lys Asn Gln Val
65 70 75 80 Val Leu Lys Val Thr Asn Met Glu Pro Ala Asp Thr Ala Thr
Tyr Tyr 85 90 95 Cys Ala Arg Ser Met Ile Thr Asn Trp Tyr Phe Asp
Val Trp Gly Ala 100 105 110 Gly Thr Thr Val Thr Val Ser Ser Ala Ser
Thr Lys Gly Pro Ser Val 115 120 125 Phe Pro Leu Ala Pro Ser Ser Lys
Ser Thr Ser Gly Gly Thr Ala Ala 130 135 140 Leu Gly Cys Leu Val Lys
Asp Tyr Phe Pro Glu Pro Val Thr Val Ser 145 150 155 160 Trp Asn Ser
Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val 165 170 175 Leu
Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro 180 185
190 Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys
195 200 205 Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser
Cys Asp 210 215 220 Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu
Ala Ala Gly Gly 225 230 235 240 Pro Ser Val Phe Leu Phe Pro Pro Lys
Pro Lys Asp Thr Leu Met Ile 245 250 255 Ser Arg Thr Pro Glu Val Thr
Cys Val Val Val Asp Val Ser His Glu 260 265 270 Asp Pro Glu Val Lys
Phe Asn Trp Tyr Val Asp Gly Val Glu Val His 275 280 285 Asn Ala Lys
Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg 290 295 300 Val
Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys 305 310
315 320 Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile
Glu 325 330 335 Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro
Gln Val Tyr 340 345 350 Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys
Asn Gln Val Ser Leu 355 360 365 Trp Cys Leu Val Lys Gly Phe Tyr Pro
Ser Asp Ile Ala Val Trp Trp 370 375 380 Glu Ser Tyr Gly Thr Glu Trp
Ser Ser Tyr Lys Thr Thr Pro Pro Val 385 390 395 400 Leu Asp Ser Asp
Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Thr 405 410 415 Lys Glu
Glu Trp Gln Gln Gly Phe Val Phe Ser Cys Ser Val Met His 420 425 430
Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro 435
440 445 Gly Lys 450 <210> SEQ ID NO 80 <211> LENGTH:
450 <212> TYPE: PRT <213> ORGANISM: Artificial Sequence
<220> FEATURE: <221> NAME/KEY: source <223> OTHER
INFORMATION: /note="Description of Artificial Sequence: Synthetic
polypeptide" <400> SEQUENCE: 80 Gln Val Thr Leu Arg 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 Thr Ser 20 25 30 Gly Met Ser
Val Gly Trp Ile Arg Gln Pro Pro Gly Lys Ala Leu Glu 35 40 45 Trp
Leu Ala Asp Ile Trp Trp Asp Asp Lys Lys Asp Tyr Asn Pro Ser 50 55
60 Leu Lys Ser Arg Leu Thr Ile Ser Lys Asp Thr Ser Lys Asn Gln Val
65 70 75 80 Val Leu Lys Val Thr Asn Met Glu Pro Ala Asp Thr Ala Thr
Tyr Tyr 85 90 95 Cys Ala Arg Ser Met Ile Thr Asn Trp Tyr Phe Asp
Val Trp Gly Ala 100 105 110 Gly Thr Thr Val Thr Val Ser Ser Ala Ser
Thr Lys Gly Pro Ser Val 115 120 125 Phe Pro Leu Ala Pro Ser Ser Lys
Ser Thr Ser Gly Gly Thr Ala Ala 130 135 140 Leu Gly Cys Leu Val Lys
Asp Tyr Phe Pro Glu Pro Val Thr Val Ser 145 150 155 160 Trp Asn Ser
Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val 165 170 175 Leu
Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro 180 185
190 Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys
195 200 205 Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser
Cys Asp 210 215 220 Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu
Ala Ala Gly Gly 225 230 235 240 Pro Ser Val Phe Leu Phe Pro Pro Lys
Pro Lys Asp Thr Leu Met Ile 245 250 255 Ser Arg Thr Pro Glu Val Thr
Cys Val Val Val Asp Val Ser His Glu 260 265 270 Asp Pro Glu Val Lys
Phe Asn Trp Tyr Val Asp Gly Val Glu Val His 275 280 285 Asn Ala Lys
Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg 290 295 300 Val
Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys 305 310
315 320 Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile
Glu 325 330 335 Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro
Gln Val Tyr 340 345 350 Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys
Asn Gln Val Ser Leu 355 360 365 Trp Cys Leu Val Lys Gly Phe Tyr Pro
Ser Asp Ile Ala Val Glu Trp 370 375 380 Glu Ser Tyr Gly Thr Glu Trp
Val Asn Tyr Lys Thr Thr Pro Pro Val 385 390 395 400 Leu Asp Ser Asp
Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Thr 405 410 415 Lys Glu
Glu Trp Gln Gln Gly Phe Val Phe Ser Cys Ser Val Met His 420 425 430
Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro 435
440 445 Gly Lys 450 <210> SEQ ID NO 81 <211> LENGTH:
450 <212> TYPE: PRT <213> ORGANISM: Artificial Sequence
<220> FEATURE: <221> NAME/KEY: source <223> OTHER
INFORMATION: /note="Description of Artificial Sequence: Synthetic
polypeptide" <400> SEQUENCE: 81 Gln Val Thr Leu Arg 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 Thr Ser 20 25 30 Gly Met Ser
Val Gly Trp Ile Arg Gln Pro Pro Gly Lys Ala Leu Glu 35 40 45 Trp
Leu Ala Asp Ile Trp Trp Asp Asp Lys Lys Asp Tyr Asn Pro Ser 50 55
60 Leu Lys Ser Arg Leu Thr Ile Ser Lys Asp Thr Ser Lys Asn Gln Val
65 70 75 80 Val Leu Lys Val Thr Asn Met Glu Pro Ala Asp Thr Ala Thr
Tyr Tyr 85 90 95 Cys Ala Arg Ser Met Ile Thr Asn Trp Tyr Phe Asp
Val Trp Gly Ala 100 105 110 Gly Thr Thr Val Thr Val Ser Ser Ala Ser
Thr Lys Gly Pro Ser Val 115 120 125 Phe Pro Leu Ala Pro Ser Ser Lys
Ser Thr Ser Gly Gly Thr Ala Ala 130 135 140 Leu Gly Cys Leu Val Lys
Asp Tyr Phe Pro Glu Pro Val Thr Val Ser 145 150 155 160 Trp Asn Ser
Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val 165 170 175 Leu
Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro 180 185
190 Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys
195 200 205 Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser
Cys Asp 210 215 220 Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu
Ala Ala Gly Gly 225 230 235 240 Pro Ser Val Phe Leu Phe Pro Pro Lys
Pro Lys Asp Thr Leu Met Ile 245 250 255 Ser Arg Thr Pro Glu Val Thr
Cys Val Val Val Asp Val Ser His Glu 260 265 270 Asp Pro Glu Val Lys
Phe Asn Trp Tyr Val Asp Gly Val Glu Val His 275 280 285 Asn Ala Lys
Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg 290 295 300 Val
Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys 305 310
315 320 Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile
Glu 325 330 335 Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro
Gln Val Tyr 340 345 350 Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys
Asn Gln Val Ser Leu 355 360 365 Ser Cys Ala Val Lys Gly Phe Tyr Pro
Ser Asp Ile Ala Val Glu Trp 370 375 380 Glu Ser Asn Gly Gln Pro Glu
Asn Asn Tyr Lys Thr Thr Pro Pro Val 385 390 395 400 Leu Asp Ser Asp
Gly Ser Phe Phe Leu Val Ser Lys Leu Thr Val Asp 405 410 415 Lys Ser
Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His 420 425 430
Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro 435
440 445 Gly Lys 450 <210> SEQ ID NO 82 <211> LENGTH:
213 <212> TYPE: PRT <213> ORGANISM: Artificial Sequence
<220> FEATURE: <221> NAME/KEY: source <223> OTHER
INFORMATION: /note="Description of Artificial Sequence: Synthetic
polypeptide" <400> SEQUENCE: 82 Asp Ile Gln Met Thr Gln Ser
Pro Ser Thr Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile
Thr Cys Lys Cys Gln Leu Ser Val Gly Tyr Met 20 25 30 His Trp Tyr
Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile Tyr 35 40 45 Asp
Thr Ser Lys Leu Ala Ser Gly Val Pro Ser Arg Phe Ser Gly Ser 50 55
60 Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro Asp
65 70 75 80 Asp Phe Ala Thr Tyr Tyr Cys Phe Gln Gly Ser Gly Tyr Pro
Phe Thr 85 90 95 Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys Arg Thr
Val Ala Ala Pro 100 105 110 Ser Val Phe Ile Phe Pro Pro Ser Asp Glu
Gln Leu Lys Ser Gly Thr 115 120 125 Ala Ser Val Val Cys Leu Leu Asn
Asn Phe Tyr Pro Arg Glu Ala Lys 130 135 140 Val Gln Trp Lys Val Asp
Asn Ala Leu Gln Ser Gly Asn Ser Gln Glu 145 150 155 160 Ser Val Thr
Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser Ser 165 170 175 Thr
Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr Ala 180 185
190 Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser Phe
195 200 205 Asn Arg Gly Glu Cys 210 <210> SEQ ID NO 83
<211> LENGTH: 448 <212> TYPE: PRT <213> ORGANISM:
Artificial Sequence <220> FEATURE: <221> NAME/KEY:
source <223> OTHER INFORMATION: /note="Description of
Artificial Sequence: Synthetic polypeptide" <400> SEQUENCE:
83 Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly
1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Thr
Asp Phe 20 25 30 Tyr Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly
Leu Glu Trp Val 35 40 45 Ser Val Ile Arg Asn Lys Ala Asn Ala Tyr
Thr Ala Gly Tyr Asn Pro 50 55 60 Ser Val Lys Gly Arg Phe Thr Ile
Ser Arg Asp Asn Ser Lys Asn Thr 65 70 75 80 Leu Tyr Leu Gln Met Asn
Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr 85 90 95 Tyr Cys Ala Arg
Leu Thr Tyr Gly Phe Asp Tyr Trp Gly Gln Gly Thr 100 105 110 Leu Val
Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro 115 120 125
Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly 130
135 140 Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp
Asn 145 150 155 160 Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro
Ala Val Leu Gln 165 170 175 Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val
Val Thr Val Pro Ser Ser 180 185 190 Ser Leu Gly Thr Gln Thr Tyr Ile
Cys Asn Val Asn His Lys Pro Ser 195 200 205 Asn Thr Lys Val Asp Lys
Lys Val Glu Pro Lys Ser Cys Asp Lys Thr 210 215 220 His Thr Cys Pro
Pro Cys Pro Ala Pro Glu Ala Ala Gly Gly Pro Ser 225 230 235 240 Val
Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg 245 250
255 Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro
260 265 270 Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His
Asn Ala 275 280 285 Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr
Tyr Arg Val Val 290 295 300 Ser Val Leu Thr Val Leu His Gln Asp Trp
Leu Asn Gly Lys Glu Tyr 305 310 315 320 Lys Cys Lys Val Ser Asn Lys
Ala Leu Pro Ala Pro Ile Glu Lys Thr 325 330 335 Ile Ser Lys Ala Lys
Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu 340 345 350 Pro Pro Ser
Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Thr Cys 355 360 365 Leu
Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser 370 375
380 Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp
385 390 395 400 Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val
Asp Lys Ser 405 410 415 Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser
Val Met His Glu Ala 420 425 430 Leu His Asn His Tyr Thr Gln Lys Ser
Leu Ser Leu Ser Pro Gly Lys 435 440 445 <210> SEQ ID NO 84
<211> LENGTH: 216 <212> TYPE: PRT <213> ORGANISM:
Artificial Sequence <220> FEATURE: <221> NAME/KEY:
source <223> OTHER INFORMATION: /note="Description of
Artificial Sequence: Synthetic polypeptide" <400> SEQUENCE:
84 Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys
1 5 10 15 Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr
Cys Val 20 25 30 Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys
Phe Asn Trp Tyr 35 40 45 Val Asp Gly Val Glu Val His Asn Ala Lys
Thr Lys Pro Arg Glu Glu 50 55 60 Gln Tyr Asn Ser Thr Tyr Arg Val
Val Ser Val Leu Thr Val Leu His 65 70 75 80 Gln Asp Trp Leu Asn Gly
Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys 85 90 95 Ala Leu Pro Ala
Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln 100 105 110 Pro Arg
Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu 115 120 125
Thr Lys Asn Gln Val Ser Leu Ser Cys Ala Val Lys Gly Phe Tyr Pro 130
135 140 Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn
Asn 145 150 155 160 Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly
Ser Phe Phe Leu 165 170 175 Val Ser Lys Leu Thr Val Asp Lys Ser Arg
Trp Gln Gln Gly Asn Val 180 185 190 Phe Ser Cys Ser Val Leu His Glu
Ala Leu His Ser His Tyr Thr Gln 195 200 205 Lys Ser Leu Ser Leu Ser
Pro Gly 210 215
1 SEQUENCE LISTING <160> NUMBER OF SEQ ID NOS: 84 <210>
SEQ ID NO 1 <211> LENGTH: 230 <212> TYPE: PRT
<213> ORGANISM: Homo sapiens <400> SEQUENCE: 1 Met Glu
Pro Leu Arg Leu Leu Ile Leu Leu Phe Val Thr Glu Leu Ser 1 5 10 15
Gly Ala His Asn Thr Thr Val Phe Gln Gly Val Ala Gly Gln Ser Leu 20
25 30 Gln Val Ser Cys Pro Tyr Asp Ser Met Lys His Trp Gly Arg Arg
Lys 35 40 45 Ala Trp Cys Arg Gln Leu Gly Glu Lys Gly Pro Cys Gln
Arg Val Val 50 55 60 Ser Thr His Asn Leu Trp Leu Leu Ser Phe Leu
Arg Arg Trp Asn Gly 65 70 75 80 Ser Thr Ala Ile Thr Asp Asp Thr Leu
Gly Gly Thr Leu Thr Ile Thr 85 90 95 Leu Arg Asn Leu Gln Pro His
Asp Ala Gly Leu Tyr Gln Cys Gln Ser 100 105 110 Leu His Gly Ser Glu
Ala Asp Thr Leu Arg Lys Val Leu Val Glu Val 115 120 125 Leu Ala Asp
Pro Leu Asp His Arg Asp Ala Gly Asp Leu Trp Phe Pro 130 135 140 Gly
Glu Ser Glu Ser Phe Glu Asp Ala His Val Glu His Ser Ile Ser 145 150
155 160 Arg Ser Leu Leu Glu Gly Glu Ile Pro Phe Pro Pro Thr Ser Ile
Leu 165 170 175 Leu Leu Leu Ala Cys Ile Phe Leu Ile Lys Ile Leu Ala
Ala Ser Ala 180 185 190 Leu Trp Ala Ala Ala Trp His Gly Gln Lys Pro
Gly Thr His Pro Pro 195 200 205 Ser Glu Leu Asp Cys Gly His Asp Pro
Gly Tyr Gln Leu Gln Thr Leu 210 215 220 Pro Gly Leu Arg Asp Thr 225
230 <210> SEQ ID NO 2 <211> LENGTH: 118 <212>
TYPE: PRT <213> ORGANISM: Artificial Sequence <220>
FEATURE: <221> NAME/KEY: source <223> OTHER
INFORMATION: /note="Description of Artificial Sequence: Synthetic
polypeptide" <400> SEQUENCE: 2 Glu Val Lys Leu Leu Asp Ser
Gly Gly Gly Leu Val Gln Ala Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser
Cys Ala Gly Ser Gly Phe Thr Phe Thr Asp Phe 20 25 30 Tyr Met Ser
Trp Ile Arg Gln Pro Pro Gly Lys Ala Pro Glu Trp Leu 35 40 45 Gly
Val Ile Arg Asn Lys Ala Asn Gly Tyr Thr Ala Gly Tyr Asn Pro 50 55
60 Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Thr Gln Asn Ile
65 70 75 80 Leu Tyr Leu Gln Met Asn Thr Leu Arg Ala Glu Asp Thr Ala
Ile Tyr 85 90 95 Tyr Cys Ala Arg Leu Ser Tyr Gly Phe Asp Tyr Trp
Gly Gln Gly Val 100 105 110 Met Val Thr Val Ser Ser 115 <210>
SEQ ID NO 3 <211> LENGTH: 112 <212> TYPE: PRT
<213> ORGANISM: Artificial Sequence <220> FEATURE:
<221> NAME/KEY: source <223> OTHER INFORMATION:
/note="Description of Artificial Sequence: Synthetic polypeptide"
<400> SEQUENCE: 3 Asp Ile Val Met Thr Gln Gly Ala Leu Pro Asn
Pro Val Pro Ser Gly 1 5 10 15 Glu Ser Ala Ser Ile Thr Cys Gln Ser
Ser Lys Ser Leu Leu His Ser 20 25 30 Asn Gly Lys Thr Tyr Leu Asn
Trp Tyr Leu Gln Arg Pro Gly Gln Ser 35 40 45 Pro Gln Leu Leu Ile
Tyr Trp Met Ser Thr Arg Ala Ser Gly Val Ser 50 55 60 Asp Arg Phe
Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Lys Ile 65 70 75 80 Ser
Ser Val Glu Ala Glu Asp Val Gly Val Tyr Tyr Cys Gln Gln Phe 85 90
95 Leu Glu Phe Pro Phe Thr Phe Gly Ser Gly Thr Lys Leu Glu Ile Lys
100 105 110 <210> SEQ ID NO 4 <211> LENGTH: 10
<212> TYPE: PRT <213> ORGANISM: Artificial Sequence
<220> FEATURE: <221> NAME/KEY: source <223> OTHER
INFORMATION: /note="Description of Artificial Sequence: Synthetic
peptide" <400> SEQUENCE: 4 Gly Phe Thr Phe Thr Asp Phe Tyr
Met Ser 1 5 10 <210> SEQ ID NO 5 <211> LENGTH: 19
<212> TYPE: PRT <213> ORGANISM: Artificial Sequence
<220> FEATURE: <221> NAME/KEY: source <223> OTHER
INFORMATION: /note="Description of Artificial Sequence: Synthetic
peptide" <400> SEQUENCE: 5 Val Ile Arg Asn Lys Ala Asn Gly
Tyr Thr Ala Gly Tyr Asn Pro Ser 1 5 10 15 Val Lys Gly <210>
SEQ ID NO 6 <211> LENGTH: 9 <212> TYPE: PRT <213>
ORGANISM: Artificial Sequence <220> FEATURE: <221>
NAME/KEY: source <223> OTHER INFORMATION: /note="Description
of Artificial Sequence: Synthetic peptide" <400> SEQUENCE: 6
Ala Arg Leu Ser Tyr Gly Phe Asp Tyr 1 5 <210> SEQ ID NO 7
<211> LENGTH: 16 <212> TYPE: PRT <213> ORGANISM:
Artificial Sequence <220> FEATURE: <221> NAME/KEY:
source <223> OTHER INFORMATION: /note="Description of
Artificial Sequence: Synthetic peptide" <400> SEQUENCE: 7 Gln
Ser Ser Lys Ser Leu Leu His Ser Asn Gly Lys Thr Tyr Leu Asn 1 5 10
15 <210> SEQ ID NO 8 <211> LENGTH: 7 <212> TYPE:
PRT <213> ORGANISM: Artificial Sequence <220> FEATURE:
<221> NAME/KEY: source <223> OTHER INFORMATION:
/note="Description of Artificial Sequence: Synthetic peptide"
<400> SEQUENCE: 8 Trp Met Ser Thr Arg Ala Ser 1 5 <210>
SEQ ID NO 9 <211> LENGTH: 9 <212> TYPE: PRT <213>
ORGANISM: Artificial Sequence <220> FEATURE: <221>
NAME/KEY: source <223> OTHER INFORMATION: /note="Description
of Artificial Sequence: Synthetic peptide" <400> SEQUENCE: 9
Gln Gln Phe Leu Glu Phe Pro Phe Thr 1 5 <210> SEQ ID NO 10
<211> LENGTH: 118 <212> TYPE: PRT <213> ORGANISM:
Artificial Sequence <220> FEATURE: <221> NAME/KEY:
source <223> OTHER INFORMATION: /note="Description of
Artificial Sequence: Synthetic polypeptide" <400> SEQUENCE:
10 Glu Val Lys Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly
1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Thr
Asn Phe 20 25 30 Tyr Met Ser Trp Ile Arg Gln Pro Pro Gly Arg Ala
Pro Glu Trp Leu 35 40 45 Gly Val Ile Arg Asn Arg Pro Asn Gly Tyr
Thr Thr Asp Tyr Asn Pro 50 55 60 Ser Val Lys Gly Arg Phe Thr Ile
Ser Arg Asp Asn Thr Gln Asn Ile 65 70 75 80 Leu Tyr Leu Gln Met Ser
Thr Leu Arg Ala Asp Asp Thr Ala Phe Tyr 85 90 95 Tyr Cys Thr Arg
Leu Thr Tyr Gly Phe Asp Tyr Trp Gly Gln Gly Val
100 105 110 Met Val Thr Val Ser Ser 115 <210> SEQ ID NO 11
<211> LENGTH: 112 <212> TYPE: PRT <213> ORGANISM:
Artificial Sequence <220> FEATURE: <221> NAME/KEY:
source <223> OTHER INFORMATION: /note="Description of
Artificial Sequence: Synthetic polypeptide" <400> SEQUENCE:
11 Asp Ile Val Met Thr Gln Gly Ala Leu Pro Asn Pro Val Pro Ser Gly
1 5 10 15 Glu Ser Ala Ser Ile Thr Cys Gln Ser Ser Lys Ser Leu Leu
His Ser 20 25 30 Asn Gly Lys Thr Tyr Leu Asn Trp Tyr Leu Gln Arg
Pro Gly Gln Ser 35 40 45 Pro Gln Leu Leu Ile Tyr Trp Met Ser Thr
Arg Ala Ser Gly Val Ser 50 55 60 Asp Arg Phe Ser Gly Ser Gly Ser
Gly Thr Asp Phe Thr Leu Lys Ile 65 70 75 80 Ser Ser Val Glu Ala Glu
Val Val Gly Val Tyr Tyr Cys Gln Gln Phe 85 90 95 Leu Glu Phe Pro
Phe Thr Phe Gly Ser Gly Thr Lys Leu Glu Ile Lys 100 105 110
<210> SEQ ID NO 12 <211> LENGTH: 10 <212> TYPE:
PRT <213> ORGANISM: Artificial Sequence <220> FEATURE:
<221> NAME/KEY: source <223> OTHER INFORMATION:
/note="Description of Artificial Sequence: Synthetic peptide"
<400> SEQUENCE: 12 Gly Phe Thr Phe Thr Asn Phe Tyr Met Ser 1
5 10 <210> SEQ ID NO 13 <211> LENGTH: 19 <212>
TYPE: PRT <213> ORGANISM: Artificial Sequence <220>
FEATURE: <221> NAME/KEY: source <223> OTHER
INFORMATION: /note="Description of Artificial Sequence: Synthetic
peptide" <400> SEQUENCE: 13 Val Ile Arg Asn Arg Pro Asn Gly
Tyr Thr Thr Asp Tyr Asn Pro Ser 1 5 10 15 Val Lys Gly <210>
SEQ ID NO 14 <211> LENGTH: 9 <212> TYPE: PRT
<213> ORGANISM: Artificial Sequence <220> FEATURE:
<221> NAME/KEY: source <223> OTHER INFORMATION:
/note="Description of Artificial Sequence: Synthetic peptide"
<400> SEQUENCE: 14 Thr Arg Leu Thr Tyr Gly Phe Asp Tyr 1 5
<210> SEQ ID NO 15 <211> LENGTH: 118 <212> TYPE:
PRT <213> ORGANISM: Artificial Sequence <220> FEATURE:
<221> NAME/KEY: source <223> OTHER INFORMATION:
/note="Description of Artificial Sequence: Synthetic polypeptide"
<400> SEQUENCE: 15 Glu Val Lys Leu Leu Asp Ser Gly Gly Gly
Leu Val Gln Ala Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Gly
Ser Gly Phe Thr Phe Thr Asp Phe 20 25 30 Tyr Met Ser Trp Ile Arg
Gln Pro Pro Gly Lys Ala Pro Glu Trp Leu 35 40 45 Gly Val Ile Arg
Asn Lys Ala Asn Gly Tyr Thr Ala Gly Tyr Asn Pro 50 55 60 Ser Val
Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Thr Gln Asn Ile 65 70 75 80
Leu Tyr Leu Gln Met Asn Thr Leu Arg Ala Glu Asp Thr Ala Ile Tyr 85
90 95 Tyr Cys Ala Arg Leu Thr Tyr Gly Phe Asp Tyr Trp Gly Gln Gly
Val 100 105 110 Met Val Thr Val Ser Ser 115 <210> SEQ ID NO
16 <211> LENGTH: 112 <212> TYPE: PRT <213>
ORGANISM: Artificial Sequence <220> FEATURE: <221>
NAME/KEY: source <223> OTHER INFORMATION: /note="Description
of Artificial Sequence: Synthetic polypeptide" <400>
SEQUENCE: 16 Asp Ile Val Met Thr Gln Gly Ala Leu Pro Asn Pro Val
Pro Ser Gly 1 5 10 15 Glu Ser Ala Ser Ile Thr Cys Gln Ser Ser Lys
Ser Leu Leu His Ser 20 25 30 Asn Gly Lys Thr Tyr Leu Asn Trp Tyr
Leu Gln Arg Pro Gly Gln Ser 35 40 45 Pro Gln Leu Leu Ile Tyr Trp
Met Ser Thr Arg Ala Ser Gly Val Ser 50 55 60 Asp Arg Phe Ser Gly
Ser Gly Ser Gly Thr Asp Phe Thr Leu Lys Ile 65 70 75 80 Ser Ser Val
Glu Ala Glu Asp Val Gly Val Tyr Tyr Cys Gln Gln Phe 85 90 95 Leu
Glu Tyr Pro Phe Thr Phe Gly Ser Gly Thr Lys Leu Glu Ile Lys 100 105
110 <210> SEQ ID NO 17 <211> LENGTH: 9 <212>
TYPE: PRT <213> ORGANISM: Artificial Sequence <220>
FEATURE: <221> NAME/KEY: source <223> OTHER
INFORMATION: /note="Description of Artificial Sequence: Synthetic
peptide" <400> SEQUENCE: 17 Ala Arg Leu Thr Tyr Gly Phe Asp
Tyr 1 5 <210> SEQ ID NO 18 <211> LENGTH: 9 <212>
TYPE: PRT <213> ORGANISM: Artificial Sequence <220>
FEATURE: <221> NAME/KEY: source <223> OTHER
INFORMATION: /note="Description of Artificial Sequence: Synthetic
peptide" <400> SEQUENCE: 18 Gln Gln Phe Leu Glu Tyr Pro Phe
Thr 1 5 <210> SEQ ID NO 19 <211> LENGTH: 118
<212> TYPE: PRT <213> ORGANISM: Artificial Sequence
<220> FEATURE: <221> NAME/KEY: source <223> OTHER
INFORMATION: /note="Description of Artificial Sequence: Synthetic
polypeptide" <400> SEQUENCE: 19 Glu Val Gln Leu Val Glu Ser
Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser
Cys Ala Ala Ser Gly Phe Thr Phe Thr Asp Phe 20 25 30 Tyr Met Ser
Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser
Val Ile Arg Asn Lys Ala Asn Gly Tyr Thr Ala Gly Tyr Asn Pro 50 55
60 Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr
65 70 75 80 Leu Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala
Val Tyr 85 90 95 Tyr Cys Ala Arg Leu Thr Tyr Gly Phe Asp Tyr Trp
Gly Gln Gly Thr 100 105 110 Leu Val Thr Val Ser Ser 115 <210>
SEQ ID NO 20 <211> LENGTH: 112 <212> TYPE: PRT
<213> ORGANISM: Artificial Sequence <220> FEATURE:
<221> NAME/KEY: source <223> OTHER INFORMATION:
/note="Description of Artificial Sequence: Synthetic polypeptide"
<400> SEQUENCE: 20 Asp Ile Val Met Thr Gln Thr Pro Leu Ser
Leu Pro Val Thr Pro Gly 1 5 10 15 Glu Pro Ala Ser Ile Ser Cys Gln
Ser Ser Lys Ser Leu Leu His Ser 20 25 30 Asn Gly Lys Thr Tyr Leu
Asn Trp Tyr Leu Gln Lys Pro Gly Gln Ser 35 40 45 Pro Gln Leu Leu
Ile Tyr Trp Met Ser Thr Arg Ala Ser Gly Val Pro 50 55 60 Asp Arg
Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Lys Ile 65 70 75 80
Ser Arg Val Glu Ala Glu Asp Val Gly Val Tyr Tyr Cys Gln Gln Phe 85
90 95 Leu Glu Tyr Pro Phe Thr Phe Gly Gln Gly Thr Lys Val Glu Ile
Lys
100 105 110 <210> SEQ ID NO 21 <211> LENGTH: 118
<212> TYPE: PRT <213> ORGANISM: Artificial Sequence
<220> FEATURE: <221> NAME/KEY: source <223> OTHER
INFORMATION: /note="Description of Artificial Sequence: Synthetic
polypeptide" <400> SEQUENCE: 21 Glu Val Gln Leu Val Glu Ser
Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser
Cys Ala Gly Ser Gly Phe Thr Phe Thr Asp Phe 20 25 30 Tyr Met Ser
Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser
Val Ile Arg Asn Lys Ala Asn Gly Tyr Thr Ala Gly Tyr Asn Pro 50 55
60 Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr
65 70 75 80 Leu Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala
Val Tyr 85 90 95 Tyr Cys Ala Arg Leu Thr Tyr Gly Phe Asp Tyr Trp
Gly Gln Gly Thr 100 105 110 Leu Val Thr Val Ser Ser 115 <210>
SEQ ID NO 22 <211> LENGTH: 112 <212> TYPE: PRT
<213> ORGANISM: Artificial Sequence <220> FEATURE:
<221> NAME/KEY: source <223> OTHER INFORMATION:
/note="Description of Artificial Sequence: Synthetic polypeptide"
<400> SEQUENCE: 22 Asp Ile Val Met Thr Gln Thr Pro Leu Ser
Leu Pro Val Thr Pro Gly 1 5 10 15 Glu Pro Ala Ser Ile Ser Cys Gln
Ser Ser Lys Ser Leu Leu His Ser 20 25 30 Thr Gly Lys Thr Tyr Leu
Asn Trp Tyr Leu Gln Lys Pro Gly Gln Ser 35 40 45 Pro Gln Leu Leu
Ile Tyr Trp Met Ser Thr Arg Ala Ser Gly Val Pro 50 55 60 Asp Arg
Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Lys Ile 65 70 75 80
Ser Arg Val Glu Ala Glu Asp Val Gly Val Tyr Tyr Cys Gln Gln Phe 85
90 95 Leu Glu Tyr Pro Phe Thr Phe Gly Gln Gly Thr Lys Val Glu Ile
Lys 100 105 110 <210> SEQ ID NO 23 <211> LENGTH: 16
<212> TYPE: PRT <213> ORGANISM: Artificial Sequence
<220> FEATURE: <221> NAME/KEY: source <223> OTHER
INFORMATION: /note="Description of Artificial Sequence: Synthetic
peptide" <400> SEQUENCE: 23 Gln Ser Ser Lys Ser Leu Leu His
Ser Thr Gly Lys Thr Tyr Leu Asn 1 5 10 15 <210> SEQ ID NO 24
<211> LENGTH: 118 <212> TYPE: PRT <213> ORGANISM:
Artificial Sequence <220> FEATURE: <221> NAME/KEY:
source <223> OTHER INFORMATION: /note="Description of
Artificial Sequence: Synthetic polypeptide" <400> SEQUENCE:
24 Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly
1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Thr
Asp Phe 20 25 30 Tyr Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly
Leu Glu Trp Val 35 40 45 Ser Val Ile Arg Asn Lys Ala Asn Ala Tyr
Thr Ala Gly Tyr Asn Pro 50 55 60 Ser Val Lys Gly Arg Phe Thr Ile
Ser Arg Asp Asn Ser Lys Asn Thr 65 70 75 80 Leu Tyr Leu Gln Met Asn
Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr 85 90 95 Tyr Cys Ala Arg
Leu Thr Tyr Gly Phe Asp Tyr Trp Gly Gln Gly Thr 100 105 110 Leu Val
Thr Val Ser Ser 115 <210> SEQ ID NO 25 <211> LENGTH: 19
<212> TYPE: PRT <213> ORGANISM: Artificial Sequence
<220> FEATURE: <221> NAME/KEY: source <223> OTHER
INFORMATION: /note="Description of Artificial Sequence: Synthetic
peptide" <400> SEQUENCE: 25 Val Ile Arg Asn Lys Ala Asn Ala
Tyr Thr Ala Gly Tyr Asn Pro Ser 1 5 10 15 Val Lys Gly <210>
SEQ ID NO 26 <211> LENGTH: 118 <212> TYPE: PRT
<213> ORGANISM: Artificial Sequence <220> FEATURE:
<221> NAME/KEY: source <223> OTHER INFORMATION:
/note="Description of Artificial Sequence: Synthetic polypeptide"
<400> SEQUENCE: 26 Glu Val Gln Leu Val Glu Ser Gly Gly Gly
Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Gly
Ser Gly Phe Thr Phe Thr Asp Phe 20 25 30 Tyr Met Ser Trp Val Arg
Gln Ala Pro Gly Lys Gly Pro Glu Trp Leu 35 40 45 Ser Val Ile Arg
Asn Lys Ala Asn Ala Tyr Thr Ala Gly Tyr Asn Pro 50 55 60 Ser Val
Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr 65 70 75 80
Leu Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr 85
90 95 Tyr Cys Ala Arg Leu Thr Tyr Gly Phe Asp Tyr Trp Gly Gln Gly
Thr 100 105 110 Leu Val Thr Val Ser Ser 115 <210> SEQ ID NO
27 <211> LENGTH: 112 <212> TYPE: PRT <213>
ORGANISM: Artificial Sequence <220> FEATURE: <221>
NAME/KEY: source <223> OTHER INFORMATION: /note="Description
of Artificial Sequence: Synthetic polypeptide" <400>
SEQUENCE: 27 Asp Ile Val Met Thr Gln Ser Pro Asp Ser Leu Ala Val
Ser Leu Gly 1 5 10 15 Glu Arg Ala Thr Ile Asn Cys Gln Ser Ser Lys
Ser Leu Leu His Ser 20 25 30 Asn Gly Lys Thr Tyr Leu Asn Trp Tyr
Gln Gln Lys Pro Gly Gln Pro 35 40 45 Pro Lys Leu Leu Ile Tyr Trp
Met Ser Thr Arg Ala Ser Gly Val Pro 50 55 60 Asp Arg Phe Ser Gly
Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile 65 70 75 80 Ser Ser Leu
Gln Ala Glu Asp Val Ala Val Tyr Tyr Cys Gln Gln Phe 85 90 95 Leu
Glu Phe Pro Phe Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys 100 105
110 <210> SEQ ID NO 28 <211> LENGTH: 10 <212>
TYPE: PRT <213> ORGANISM: Artificial Sequence <220>
FEATURE: <221> NAME/KEY: source <223> OTHER
INFORMATION: /note="Description of Artificial Sequence: Synthetic
peptide" <220> FEATURE: <221> NAME/KEY: VARIANT
<222> LOCATION: (6)..(6) <223> OTHER INFORMATION:
/replace="Asn" <220> FEATURE: <221> NAME/KEY: SITE
<222> LOCATION: (1)..(10) <223> OTHER INFORMATION:
/note="Variant residues given in the sequence have no preference
with respect to those in the annotations for variant positions"
<400> SEQUENCE: 28 Gly Phe Thr Phe Thr Asp Phe Tyr Met Ser 1
5 10 <210> SEQ ID NO 29 <211> LENGTH: 19 <212>
TYPE: PRT <213> ORGANISM: Artificial Sequence <220>
FEATURE: <221> NAME/KEY: source <223> OTHER
INFORMATION: /note="Description of Artificial Sequence: Synthetic
peptide" <220> FEATURE: <221> NAME/KEY: VARIANT
<222> LOCATION: (5)..(5) <223> OTHER INFORMATION:
/replace="Arg" <220> FEATURE: <221> NAME/KEY: VARIANT
<222> LOCATION: (6)..(6) <223> OTHER INFORMATION:
/replace="Pro"
<220> FEATURE: <221> NAME/KEY: VARIANT <222>
LOCATION: (8)..(8) <223> OTHER INFORMATION: /replace="Ala"
<220> FEATURE: <221> NAME/KEY: VARIANT <222>
LOCATION: (11)..(11) <223> OTHER INFORMATION: /replace="Thr"
<220> FEATURE: <221> NAME/KEY: VARIANT <222>
LOCATION: (12)..(12) <223> OTHER INFORMATION: /replace="Asp"
<220> FEATURE: <221> NAME/KEY: SITE <222>
LOCATION: (1)..(19) <223> OTHER INFORMATION: /note="Variant
residues given in the sequence have no preference with respect to
those in the annotations for variant positions" <400>
SEQUENCE: 29 Val Ile Arg Asn Lys Ala Asn Gly Tyr Thr Ala Gly Tyr
Asn Pro Ser 1 5 10 15 Val Lys Gly <210> SEQ ID NO 30
<211> LENGTH: 9 <212> TYPE: PRT <213> ORGANISM:
Artificial Sequence <220> FEATURE: <221> NAME/KEY:
source <223> OTHER INFORMATION: /note="Description of
Artificial Sequence: Synthetic peptide" <220> FEATURE:
<221> NAME/KEY: VARIANT <222> LOCATION: (1)..(1)
<223> OTHER INFORMATION: /replace="Thr" <220> FEATURE:
<221> NAME/KEY: VARIANT <222> LOCATION: (4)..(4)
<223> OTHER INFORMATION: /replace="Ser" <220> FEATURE:
<221> NAME/KEY: SITE <222> LOCATION: (1)..(9)
<223> OTHER INFORMATION: /note="Variant residues given in the
sequence have no preference with respect to those in the
annotations for variant positions" <400> SEQUENCE: 30 Ala Arg
Leu Thr Tyr Gly Phe Asp Tyr 1 5 <210> SEQ ID NO 31
<211> LENGTH: 16 <212> TYPE: PRT <213> ORGANISM:
Artificial Sequence <220> FEATURE: <221> NAME/KEY:
source <223> OTHER INFORMATION: /note="Description of
Artificial Sequence: Synthetic peptide" <220> FEATURE:
<221> NAME/KEY: VARIANT <222> LOCATION: (10)..(10)
<223> OTHER INFORMATION: /replace="Thr" <220> FEATURE:
<221> NAME/KEY: SITE <222> LOCATION: (1)..(16)
<223> OTHER INFORMATION: /note="Variant residues given in the
sequence have no preference with respect to those in the
annotations for variant positions" <400> SEQUENCE: 31 Gln Ser
Ser Lys Ser Leu Leu His Ser Asn Gly Lys Thr Tyr Leu Asn 1 5 10 15
<210> SEQ ID NO 32 <211> LENGTH: 9 <212> TYPE:
PRT <213> ORGANISM: Artificial Sequence <220> FEATURE:
<221> NAME/KEY: source <223> OTHER INFORMATION:
/note="Description of Artificial Sequence: Synthetic peptide"
<220> FEATURE: <221> NAME/KEY: VARIANT <222>
LOCATION: (6)..(6) <223> OTHER INFORMATION: /replace="Phe"
<220> FEATURE: <221> NAME/KEY: SITE <222>
LOCATION: (1)..(9) <223> OTHER INFORMATION: /note="Variant
residues given in the sequence have no preference with respect to
those in the annotations for variant positions" <400>
SEQUENCE: 32 Gln Gln Phe Leu Glu Tyr Pro Phe Thr 1 5 <210>
SEQ ID NO 33 <400> SEQUENCE: 33 000 <210> SEQ ID NO 34
<211> LENGTH: 5 <212> TYPE: PRT <213> ORGANISM:
Artificial Sequence <220> FEATURE: <221> NAME/KEY:
source <223> OTHER INFORMATION: /note="Description of
Artificial Sequence: Synthetic peptide" <400> SEQUENCE: 34
Gly Gly Gly Gly Ser 1 5 <210> SEQ ID NO 35 <211>
LENGTH: 6 <212> TYPE: PRT <213> ORGANISM: Artificial
Sequence <220> FEATURE: <221> NAME/KEY: source
<223> OTHER INFORMATION: /note="Description of Artificial
Sequence: Synthetic 6xHis tag" <400> SEQUENCE: 35 His His His
His His His 1 5 <210> SEQ ID NO 36 <211> LENGTH: 113
<212> TYPE: PRT <213> ORGANISM: Artificial Sequence
<220> FEATURE: <221> NAME/KEY: source <223> OTHER
INFORMATION: /note="Description of Artificial Sequence: Synthetic
polypeptide" <400> SEQUENCE: 36 Ala Ser Thr Lys Gly Pro Ser
Val Phe Pro Leu Ala Pro Ser Ser Lys 1 5 10 15 Ser Thr Ser Gly Gly
Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr 20 25 30 Phe Pro Glu
Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser 35 40 45 Gly
Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser 50 55
60 Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr
65 70 75 80 Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val
Asp Lys 85 90 95 Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr
Cys Pro Pro Cys 100 105 110 Pro <210> SEQ ID NO 37
<211> LENGTH: 107 <212> TYPE: PRT <213> ORGANISM:
Artificial Sequence <220> FEATURE: <221> NAME/KEY:
source <223> OTHER INFORMATION: /note="Description of
Artificial Sequence: Synthetic polypeptide" <400> SEQUENCE:
37 Arg Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu
1 5 10 15 Gln Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn
Asn Phe 20 25 30 Tyr Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp
Asn Ala Leu Gln 35 40 45 Ser Gly Asn Ser Gln Glu Ser Val Thr Glu
Gln Asp Ser Lys Asp Ser 50 55 60 Thr Tyr Ser Leu Ser Ser Thr Leu
Thr Leu Ser Lys Ala Asp Tyr Glu 65 70 75 80 Lys His Lys Val Tyr Ala
Cys Glu Val Thr His Gln Gly Leu Ser Ser 85 90 95 Pro Val Thr Lys
Ser Phe Asn Arg Gly Glu Cys 100 105 <210> SEQ ID NO 38
<211> LENGTH: 217 <212> TYPE: PRT <213> ORGANISM:
Homo sapiens <400> SEQUENCE: 38 Ala Pro Glu Leu Leu Gly Gly
Pro Ser Val Phe Leu Phe Pro Pro Lys 1 5 10 15 Pro Lys Asp Thr Leu
Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val 20 25 30 Val Val Asp
Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr 35 40 45 Val
Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu 50 55
60 Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His
65 70 75 80 Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser
Asn Lys 85 90 95 Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys
Ala Lys Gly Gln 100 105 110 Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro
Pro Ser Arg Asp Glu Leu 115 120 125 Thr Lys Asn Gln Val Ser Leu Thr
Cys Leu Val Lys Gly Phe Tyr Pro 130 135 140 Ser Asp Ile Ala Val Glu
Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn 145 150 155 160
Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu 165
170 175 Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn
Val 180 185 190 Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His
Tyr Thr Gln 195 200 205 Lys Ser Leu Ser Leu Ser Pro Gly Lys 210 215
<210> SEQ ID NO 39 <211> LENGTH: 217 <212> TYPE:
PRT <213> ORGANISM: Artificial Sequence <220> FEATURE:
<221> NAME/KEY: source <223> OTHER INFORMATION:
/note="Description of Artificial Sequence: Synthetic polypeptide"
<400> SEQUENCE: 39 Ala Pro Glu Ala Ala Gly Gly Pro Ser Val
Phe Leu Phe Pro Pro Lys 1 5 10 15 Pro Lys Asp Thr Leu Met Ile Ser
Arg Thr Pro Glu Val Thr Cys Val 20 25 30 Val Val Asp Val Ser His
Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr 35 40 45 Val Asp Gly Val
Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu 50 55 60 Gln Tyr
Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His 65 70 75 80
Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys 85
90 95 Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly
Gln 100 105 110 Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg
Asp Glu Leu 115 120 125 Thr Lys Asn Gln Val Ser Leu Ser Cys Ala Val
Lys Gly Phe Tyr Pro 130 135 140 Ser Asp Ile Ala Val Glu Trp Glu Ser
Asn Gly Gln Pro Glu Asn Asn 145 150 155 160 Tyr Lys Thr Thr Pro Pro
Val Leu Asp Ser Asp Gly Ser Phe Phe Leu 165 170 175 Val Ser Lys Leu
Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val 180 185 190 Phe Ser
Cys Ser Val Leu His Glu Ala Leu His Ser His Tyr Thr Gln 195 200 205
Lys Ser Leu Ser Leu Ser Pro Gly Lys 210 215 <210> SEQ ID NO
40 <211> LENGTH: 217 <212> TYPE: PRT <213>
ORGANISM: Artificial Sequence <220> FEATURE: <221>
NAME/KEY: source <223> OTHER INFORMATION: /note="Description
of Artificial Sequence: Synthetic polypeptide" <400>
SEQUENCE: 40 Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe
Pro Pro Lys 1 5 10 15 Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro
Glu Val Thr Cys Val 20 25 30 Val Val Asp Val Ser His Glu Asp Pro
Glu Val Lys Phe Asn Trp Tyr 35 40 45 Val Asp Gly Val Glu Val His
Asn Ala Lys Thr Lys Pro Arg Glu Glu 50 55 60 Gln Tyr Asn Ser Thr
Tyr Arg Val Val Ser Val Leu Thr Val Leu His 65 70 75 80 Gln Asp Trp
Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys 85 90 95 Ala
Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln 100 105
110 Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu
115 120 125 Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe
Tyr Pro 130 135 140 Ser Asp Ile Ala Val Trp Trp Glu Ser Tyr Gly Thr
Glu Trp Ser Ser 145 150 155 160 Tyr Lys Thr Thr Pro Pro Val Leu Asp
Ser Asp Gly Ser Phe Phe Leu 165 170 175 Tyr Ser Lys Leu Thr Val Thr
Lys Glu Glu Trp Gln Gln Gly Phe Val 180 185 190 Phe Ser Cys Ser Val
Met His Glu Ala Leu His Asn His Tyr Thr Gln 195 200 205 Lys Ser Leu
Ser Leu Ser Pro Gly Lys 210 215 <210> SEQ ID NO 41
<211> LENGTH: 217 <212> TYPE: PRT <213> ORGANISM:
Artificial Sequence <220> FEATURE: <221> NAME/KEY:
source <223> OTHER INFORMATION: /note="Description of
Artificial Sequence: Synthetic polypeptide" <400> SEQUENCE:
41 Ala Pro Glu Ala Ala Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys
1 5 10 15 Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr
Cys Val 20 25 30 Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys
Phe Asn Trp Tyr 35 40 45 Val Asp Gly Val Glu Val His Asn Ala Lys
Thr Lys Pro Arg Glu Glu 50 55 60 Gln Tyr Asn Ser Thr Tyr Arg Val
Val Ser Val Leu Thr Val Leu His 65 70 75 80 Gln Asp Trp Leu Asn Gly
Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys 85 90 95 Ala Leu Pro Ala
Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln 100 105 110 Pro Arg
Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu 115 120 125
Thr Lys Asn Gln Val Ser Leu Trp Cys Leu Val Lys Gly Phe Tyr Pro 130
135 140 Ser Asp Ile Ala Val Trp Trp Glu Ser Tyr Gly Thr Glu Trp Ser
Ser 145 150 155 160 Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly
Ser Phe Phe Leu 165 170 175 Tyr Ser Lys Leu Thr Val Thr Lys Glu Glu
Trp Gln Gln Gly Phe Val 180 185 190 Phe Ser Cys Ser Val Leu His Glu
Ala Leu His Ser His Tyr Thr Gln 195 200 205 Lys Ser Leu Ser Leu Ser
Pro Gly Lys 210 215 <210> SEQ ID NO 42 <211> LENGTH:
448 <212> TYPE: PRT <213> ORGANISM: Artificial Sequence
<220> FEATURE: <221> NAME/KEY: source <223> OTHER
INFORMATION: /note="Description of Artificial Sequence: Synthetic
polypeptide" <400> SEQUENCE: 42 Glu Val Gln Leu Val Glu Ser
Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser
Cys Ala Ala Ser Gly Phe Thr Phe Thr Asp Phe 20 25 30 Tyr Met Ser
Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser
Val Ile Arg Asn Lys Ala Asn Ala Tyr Thr Ala Gly Tyr Asn Pro 50 55
60 Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr
65 70 75 80 Leu Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala
Val Tyr 85 90 95 Tyr Cys Ala Arg Leu Thr Tyr Gly Phe Asp Tyr Trp
Gly Gln Gly Thr 100 105 110 Leu Val Thr Val Ser Ser Ala Ser Thr Lys
Gly Pro Ser Val Phe Pro 115 120 125 Leu Ala Pro Ser Ser Lys Ser Thr
Ser Gly Gly Thr Ala Ala Leu Gly 130 135 140 Cys Leu Val Lys Asp Tyr
Phe Pro Glu Pro Val Thr Val Ser Trp Asn 145 150 155 160 Ser Gly Ala
Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln 165 170 175 Ser
Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser 180 185
190 Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser
195 200 205 Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp
Lys Thr 210 215 220 His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala
Gly Gly Pro Ser 225 230 235 240 Val Phe Leu Phe Pro Pro Lys Pro Lys
Asp Thr Leu Met Ile Ser Arg 245 250 255 Thr Pro Glu Val Thr Cys Val
Val Val Asp Val Ser His Glu Asp Pro 260 265 270 Glu Val Lys Phe Asn
Trp Tyr Val Asp Gly Val Glu Val His Asn Ala 275 280 285 Lys Thr Lys
Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val 290 295 300 Ser
Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr 305 310
315 320 Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys
Thr 325 330 335 Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val
Tyr Thr Leu
340 345 350 Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu
Trp Cys 355 360 365 Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val
Trp Trp Glu Ser 370 375 380 Tyr Gly Thr Glu Trp Ser Ser Tyr Lys Thr
Thr Pro Pro Val Leu Asp 385 390 395 400 Ser Asp Gly Ser Phe Phe Leu
Tyr Ser Lys Leu Thr Val Thr Lys Glu 405 410 415 Glu Trp Gln Gln Gly
Phe Val Phe Ser Cys Ser Val Leu His Glu Ala 420 425 430 Leu His Ser
His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys 435 440 445
<210> SEQ ID NO 43 <211> LENGTH: 217 <212> TYPE:
PRT <213> ORGANISM: Artificial Sequence <220> FEATURE:
<221> NAME/KEY: source <223> OTHER INFORMATION:
/note="Description of Artificial Sequence: Synthetic polypeptide"
<400> SEQUENCE: 43 Ala Pro Glu Leu Leu Gly Gly Pro Ser Val
Phe Leu Phe Pro Pro Lys 1 5 10 15 Pro Lys Asp Thr Leu Met Ile Ser
Arg Thr Pro Glu Val Thr Cys Val 20 25 30 Val Val Asp Val Ser His
Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr 35 40 45 Val Asp Gly Val
Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu 50 55 60 Gln Tyr
Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His 65 70 75 80
Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys 85
90 95 Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly
Gln 100 105 110 Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg
Asp Glu Leu 115 120 125 Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val
Lys Gly Phe Tyr Pro 130 135 140 Ser Asp Ile Ala Val Glu Trp Glu Ser
Phe Gly Thr Glu Trp Ser Asn 145 150 155 160 Tyr Lys Thr Thr Pro Pro
Val Leu Asp Ser Asp Gly Ser Phe Phe Leu 165 170 175 Tyr Ser Lys Leu
Thr Val Ser Lys Glu Glu Trp Gln Gln Gly Phe Val 180 185 190 Phe Ser
Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln 195 200 205
Lys Ser Leu Ser Leu Ser Pro Gly Lys 210 215 <210> SEQ ID NO
44 <211> LENGTH: 217 <212> TYPE: PRT <213>
ORGANISM: Artificial Sequence <220> FEATURE: <221>
NAME/KEY: source <223> OTHER INFORMATION: /note="Description
of Artificial Sequence: Synthetic polypeptide" <400>
SEQUENCE: 44 Ala Pro Glu Ala Ala Gly Gly Pro Ser Val Phe Leu Phe
Pro Pro Lys 1 5 10 15 Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro
Glu Val Thr Cys Val 20 25 30 Val Val Asp Val Ser His Glu Asp Pro
Glu Val Lys Phe Asn Trp Tyr 35 40 45 Val Asp Gly Val Glu Val His
Asn Ala Lys Thr Lys Pro Arg Glu Glu 50 55 60 Gln Tyr Asn Ser Thr
Tyr Arg Val Val Ser Val Leu Thr Val Leu His 65 70 75 80 Gln Asp Trp
Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys 85 90 95 Ala
Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln 100 105
110 Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu
115 120 125 Thr Lys Asn Gln Val Ser Leu Trp Cys Leu Val Lys Gly Phe
Tyr Pro 130 135 140 Ser Asp Ile Ala Val Glu Trp Glu Ser Phe Gly Thr
Glu Trp Ser Asn 145 150 155 160 Tyr Lys Thr Thr Pro Pro Val Leu Asp
Ser Asp Gly Ser Phe Phe Leu 165 170 175 Tyr Ser Lys Leu Thr Val Ser
Lys Glu Glu Trp Gln Gln Gly Phe Val 180 185 190 Phe Ser Cys Ser Val
Leu His Glu Ala Leu His Ser His Tyr Thr Gln 195 200 205 Lys Ser Leu
Ser Leu Ser Pro Gly Lys 210 215 <210> SEQ ID NO 45
<211> LENGTH: 448 <212> TYPE: PRT <213> ORGANISM:
Artificial Sequence <220> FEATURE: <221> NAME/KEY:
source <223> OTHER INFORMATION: /note="Description of
Artificial Sequence: Synthetic polypeptide" <400> SEQUENCE:
45 Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly
1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Thr
Asp Phe 20 25 30 Tyr Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly
Leu Glu Trp Val 35 40 45 Ser Val Ile Arg Asn Lys Ala Asn Ala Tyr
Thr Ala Gly Tyr Asn Pro 50 55 60 Ser Val Lys Gly Arg Phe Thr Ile
Ser Arg Asp Asn Ser Lys Asn Thr 65 70 75 80 Leu Tyr Leu Gln Met Asn
Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr 85 90 95 Tyr Cys Ala Arg
Leu Thr Tyr Gly Phe Asp Tyr Trp Gly Gln Gly Thr 100 105 110 Leu Val
Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro 115 120 125
Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly 130
135 140 Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp
Asn 145 150 155 160 Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro
Ala Val Leu Gln 165 170 175 Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val
Val Thr Val Pro Ser Ser 180 185 190 Ser Leu Gly Thr Gln Thr Tyr Ile
Cys Asn Val Asn His Lys Pro Ser 195 200 205 Asn Thr Lys Val Asp Lys
Lys Val Glu Pro Lys Ser Cys Asp Lys Thr 210 215 220 His Thr Cys Pro
Pro Cys Pro Ala Pro Glu Ala Ala Gly Gly Pro Ser 225 230 235 240 Val
Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg 245 250
255 Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro
260 265 270 Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His
Asn Ala 275 280 285 Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr
Tyr Arg Val Val 290 295 300 Ser Val Leu Thr Val Leu His Gln Asp Trp
Leu Asn Gly Lys Glu Tyr 305 310 315 320 Lys Cys Lys Val Ser Asn Lys
Ala Leu Pro Ala Pro Ile Glu Lys Thr 325 330 335 Ile Ser Lys Ala Lys
Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu 340 345 350 Pro Pro Ser
Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Trp Cys 355 360 365 Leu
Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser 370 375
380 Phe Gly Thr Glu Trp Ser Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp
385 390 395 400 Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val
Ser Lys Glu 405 410 415 Glu Trp Gln Gln Gly Phe Val Phe Ser Cys Ser
Val Leu His Glu Ala 420 425 430 Leu His Ser His Tyr Thr Gln Lys Ser
Leu Ser Leu Ser Pro Gly Lys 435 440 445 <210> SEQ ID NO 46
<211> LENGTH: 217 <212> TYPE: PRT <213> ORGANISM:
Artificial Sequence <220> FEATURE: <221> NAME/KEY:
source <223> OTHER INFORMATION: /note="Description of
Artificial Sequence: Synthetic polypeptide" <400> SEQUENCE:
46 Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys
1 5 10 15 Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr
Cys Val 20 25 30 Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys
Phe Asn Trp Tyr 35 40 45 Val Asp Gly Val Glu Val His Asn Ala Lys
Thr Lys Pro Arg Glu Glu 50 55 60 Gln Tyr Asn Ser Thr Tyr Arg Val
Val Ser Val Leu Thr Val Leu His 65 70 75 80
Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys 85
90 95 Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly
Gln 100 105 110 Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg
Asp Glu Leu 115 120 125 Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val
Lys Gly Phe Tyr Pro 130 135 140 Ser Asp Ile Ala Val Glu Trp Glu Ser
Tyr Gly Thr Glu Trp Val Asn 145 150 155 160 Tyr Lys Thr Thr Pro Pro
Val Leu Asp Ser Asp Gly Ser Phe Phe Leu 165 170 175 Tyr Ser Lys Leu
Thr Val Thr Lys Glu Glu Trp Gln Gln Gly Phe Val 180 185 190 Phe Ser
Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln 195 200 205
Lys Ser Leu Ser Leu Ser Pro Gly Lys 210 215 <210> SEQ ID NO
47 <211> LENGTH: 217 <212> TYPE: PRT <213>
ORGANISM: Artificial Sequence <220> FEATURE: <221>
NAME/KEY: source <223> OTHER INFORMATION: /note="Description
of Artificial Sequence: Synthetic polypeptide" <400>
SEQUENCE: 47 Ala Pro Glu Ala Ala Gly Gly Pro Ser Val Phe Leu Phe
Pro Pro Lys 1 5 10 15 Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro
Glu Val Thr Cys Val 20 25 30 Val Val Asp Val Ser His Glu Asp Pro
Glu Val Lys Phe Asn Trp Tyr 35 40 45 Val Asp Gly Val Glu Val His
Asn Ala Lys Thr Lys Pro Arg Glu Glu 50 55 60 Gln Tyr Asn Ser Thr
Tyr Arg Val Val Ser Val Leu Thr Val Leu His 65 70 75 80 Gln Asp Trp
Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys 85 90 95 Ala
Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln 100 105
110 Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu
115 120 125 Thr Lys Asn Gln Val Ser Leu Trp Cys Leu Val Lys Gly Phe
Tyr Pro 130 135 140 Ser Asp Ile Ala Val Glu Trp Glu Ser Tyr Gly Thr
Glu Trp Val Asn 145 150 155 160 Tyr Lys Thr Thr Pro Pro Val Leu Asp
Ser Asp Gly Ser Phe Phe Leu 165 170 175 Tyr Ser Lys Leu Thr Val Thr
Lys Glu Glu Trp Gln Gln Gly Phe Val 180 185 190 Phe Ser Cys Ser Val
Leu His Glu Ala Leu His Ser His Tyr Thr Gln 195 200 205 Lys Ser Leu
Ser Leu Ser Pro Gly Lys 210 215 <210> SEQ ID NO 48
<211> LENGTH: 448 <212> TYPE: PRT <213> ORGANISM:
Artificial Sequence <220> FEATURE: <221> NAME/KEY:
source <223> OTHER INFORMATION: /note="Description of
Artificial Sequence: Synthetic polypeptide" <400> SEQUENCE:
48 Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly
1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Thr
Asp Phe 20 25 30 Tyr Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly
Leu Glu Trp Val 35 40 45 Ser Val Ile Arg Asn Lys Ala Asn Ala Tyr
Thr Ala Gly Tyr Asn Pro 50 55 60 Ser Val Lys Gly Arg Phe Thr Ile
Ser Arg Asp Asn Ser Lys Asn Thr 65 70 75 80 Leu Tyr Leu Gln Met Asn
Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr 85 90 95 Tyr Cys Ala Arg
Leu Thr Tyr Gly Phe Asp Tyr Trp Gly Gln Gly Thr 100 105 110 Leu Val
Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro 115 120 125
Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly 130
135 140 Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp
Asn 145 150 155 160 Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro
Ala Val Leu Gln 165 170 175 Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val
Val Thr Val Pro Ser Ser 180 185 190 Ser Leu Gly Thr Gln Thr Tyr Ile
Cys Asn Val Asn His Lys Pro Ser 195 200 205 Asn Thr Lys Val Asp Lys
Lys Val Glu Pro Lys Ser Cys Asp Lys Thr 210 215 220 His Thr Cys Pro
Pro Cys Pro Ala Pro Glu Ala Ala Gly Gly Pro Ser 225 230 235 240 Val
Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg 245 250
255 Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro
260 265 270 Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His
Asn Ala 275 280 285 Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr
Tyr Arg Val Val 290 295 300 Ser Val Leu Thr Val Leu His Gln Asp Trp
Leu Asn Gly Lys Glu Tyr 305 310 315 320 Lys Cys Lys Val Ser Asn Lys
Ala Leu Pro Ala Pro Ile Glu Lys Thr 325 330 335 Ile Ser Lys Ala Lys
Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu 340 345 350 Pro Pro Ser
Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Trp Cys 355 360 365 Leu
Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser 370 375
380 Tyr Gly Thr Glu Trp Val Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp
385 390 395 400 Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val
Thr Lys Glu 405 410 415 Glu Trp Gln Gln Gly Phe Val Phe Ser Cys Ser
Val Leu His Glu Ala 420 425 430 Leu His Ser His Tyr Thr Gln Lys Ser
Leu Ser Leu Ser Pro Gly Lys 435 440 445 <210> SEQ ID NO 49
<211> LENGTH: 217 <212> TYPE: PRT <213> ORGANISM:
Artificial Sequence <220> FEATURE: <221> NAME/KEY:
source <223> OTHER INFORMATION: /note="Description of
Artificial Sequence: Synthetic polypeptide" <400> SEQUENCE:
49 Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys
1 5 10 15 Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr
Cys Val 20 25 30 Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys
Phe Asn Trp Tyr 35 40 45 Val Asp Gly Val Glu Val His Asn Ala Lys
Thr Lys Pro Arg Glu Glu 50 55 60 Gln Tyr Asn Ser Thr Tyr Arg Val
Val Ser Val Leu Thr Val Leu His 65 70 75 80 Gln Asp Trp Leu Asn Gly
Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys 85 90 95 Ala Leu Pro Ala
Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln 100 105 110 Pro Arg
Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu 115 120 125
Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro 130
135 140 Ser Asp Ile Ala Val Glu Trp Glu Ser Tyr Gly Thr Glu Trp Ser
Asn 145 150 155 160 Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly
Ser Phe Phe Leu 165 170 175 Tyr Ser Lys Leu Thr Val Ser Lys Glu Glu
Trp Gln Gln Gly Phe Val 180 185 190 Phe Ser Cys Ser Val Met His Glu
Ala Leu His Asn His Tyr Thr Gln 195 200 205 Lys Ser Leu Ser Leu Ser
Pro Gly Lys 210 215 <210> SEQ ID NO 50 <211> LENGTH:
217 <212> TYPE: PRT <213> ORGANISM: Artificial Sequence
<220> FEATURE: <221> NAME/KEY: source <223> OTHER
INFORMATION: /note="Description of Artificial Sequence: Synthetic
polypeptide" <400> SEQUENCE: 50 Ala Pro Glu Ala Ala Gly Gly
Pro Ser Val Phe Leu Phe Pro Pro Lys 1 5 10 15 Pro Lys Asp Thr Leu
Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val 20 25 30 Val Val Asp
Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr 35 40 45
Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu 50
55 60 Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu
His 65 70 75 80 Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val
Ser Asn Lys 85 90 95 Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser
Lys Ala Lys Gly Gln 100 105 110 Pro Arg Glu Pro Gln Val Tyr Thr Leu
Pro Pro Ser Arg Asp Glu Leu 115 120 125 Thr Lys Asn Gln Val Ser Leu
Trp Cys Leu Val Lys Gly Phe Tyr Pro 130 135 140 Ser Asp Ile Ala Val
Glu Trp Glu Ser Tyr Gly Thr Glu Trp Ser Asn 145 150 155 160 Tyr Lys
Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu 165 170 175
Tyr Ser Lys Leu Thr Val Ser Lys Glu Glu Trp Gln Gln Gly Phe Val 180
185 190 Phe Ser Cys Ser Val Leu His Glu Ala Leu His Ser His Tyr Thr
Gln 195 200 205 Lys Ser Leu Ser Leu Ser Pro Gly Lys 210 215
<210> SEQ ID NO 51 <211> LENGTH: 448 <212> TYPE:
PRT <213> ORGANISM: Artificial Sequence <220> FEATURE:
<221> NAME/KEY: source <223> OTHER INFORMATION:
/note="Description of Artificial Sequence: Synthetic polypeptide"
<400> SEQUENCE: 51 Glu Val Gln Leu Val Glu Ser Gly Gly Gly
Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala
Ser Gly Phe Thr Phe Thr Asp Phe 20 25 30 Tyr Met Ser Trp Val Arg
Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Val Ile Arg
Asn Lys Ala Asn Ala Tyr Thr Ala Gly Tyr Asn Pro 50 55 60 Ser Val
Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr 65 70 75 80
Leu Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr 85
90 95 Tyr Cys Ala Arg Leu Thr Tyr Gly Phe Asp Tyr Trp Gly Gln Gly
Thr 100 105 110 Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser
Val Phe Pro 115 120 125 Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly
Thr Ala Ala Leu Gly 130 135 140 Cys Leu Val Lys Asp Tyr Phe Pro Glu
Pro Val Thr Val Ser Trp Asn 145 150 155 160 Ser Gly Ala Leu Thr Ser
Gly Val His Thr Phe Pro Ala Val Leu Gln 165 170 175 Ser Ser Gly Leu
Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser 180 185 190 Ser Leu
Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser 195 200 205
Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr 210
215 220 His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Gly Pro
Ser 225 230 235 240 Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu
Met Ile Ser Arg 245 250 255 Thr Pro Glu Val Thr Cys Val Val Val Asp
Val Ser His Glu Asp Pro 260 265 270 Glu Val Lys Phe Asn Trp Tyr Val
Asp Gly Val Glu Val His Asn Ala 275 280 285 Lys Thr Lys Pro Arg Glu
Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val 290 295 300 Ser Val Leu Thr
Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr 305 310 315 320 Lys
Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr 325 330
335 Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu
340 345 350 Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu
Trp Cys 355 360 365 Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val
Glu Trp Glu Ser 370 375 380 Tyr Gly Thr Glu Trp Ser Asn Tyr Lys Thr
Thr Pro Pro Val Leu Asp 385 390 395 400 Ser Asp Gly Ser Phe Phe Leu
Tyr Ser Lys Leu Thr Val Ser Lys Glu 405 410 415 Glu Trp Gln Gln Gly
Phe Val Phe Ser Cys Ser Val Leu His Glu Ala 420 425 430 Leu His Ser
His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys 435 440 445
<210> SEQ ID NO 52 <211> LENGTH: 448 <212> TYPE:
PRT <213> ORGANISM: Artificial Sequence <220> FEATURE:
<221> NAME/KEY: source <223> OTHER INFORMATION:
/note="Description of Artificial Sequence: Synthetic polypeptide"
<400> SEQUENCE: 52 Glu Val Gln Leu Val Glu Ser Gly Gly Gly
Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala
Ser Gly Phe Thr Phe Thr Asp Phe 20 25 30 Tyr Met Ser Trp Val Arg
Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Val Ile Arg
Asn Lys Ala Asn Ala Tyr Thr Ala Gly Tyr Asn Pro 50 55 60 Ser Val
Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr 65 70 75 80
Leu Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr 85
90 95 Tyr Cys Ala Arg Leu Thr Tyr Gly Phe Asp Tyr Trp Gly Gln Gly
Thr 100 105 110 Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser
Val Phe Pro 115 120 125 Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly
Thr Ala Ala Leu Gly 130 135 140 Cys Leu Val Lys Asp Tyr Phe Pro Glu
Pro Val Thr Val Ser Trp Asn 145 150 155 160 Ser Gly Ala Leu Thr Ser
Gly Val His Thr Phe Pro Ala Val Leu Gln 165 170 175 Ser Ser Gly Leu
Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser 180 185 190 Ser Leu
Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser 195 200 205
Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr 210
215 220 His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro
Ser 225 230 235 240 Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu
Met Ile Ser Arg 245 250 255 Thr Pro Glu Val Thr Cys Val Val Val Asp
Val Ser His Glu Asp Pro 260 265 270 Glu Val Lys Phe Asn Trp Tyr Val
Asp Gly Val Glu Val His Asn Ala 275 280 285 Lys Thr Lys Pro Arg Glu
Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val 290 295 300 Ser Val Leu Thr
Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr 305 310 315 320 Lys
Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr 325 330
335 Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu
340 345 350 Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu
Ser Cys 355 360 365 Ala Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val
Glu Trp Glu Ser 370 375 380 Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr
Thr Pro Pro Val Leu Asp 385 390 395 400 Ser Asp Gly Ser Phe Phe Leu
Val Ser Lys Leu Thr Val Asp Lys Ser 405 410 415 Arg Trp Gln Gln Gly
Asn Val Phe Ser Cys Ser Val Leu His Glu Ala 420 425 430 Leu His Ser
His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys 435 440 445
<210> SEQ ID NO 53 <211> LENGTH: 448 <212> TYPE:
PRT <213> ORGANISM: Artificial Sequence <220> FEATURE:
<221> NAME/KEY: source <223> OTHER INFORMATION:
/note="Description of Artificial Sequence: Synthetic polypeptide"
<400> SEQUENCE: 53 Glu Val Gln Leu Val Glu Ser Gly Gly Gly
Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala
Ser Gly Phe Thr Phe Thr Asp Phe 20 25 30 Tyr Met Ser Trp Val Arg
Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Val Ile Arg
Asn Lys Ala Asn Ala Tyr Thr Ala Gly Tyr Asn Pro 50 55 60 Ser Val
Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr
65 70 75 80 Leu Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala
Val Tyr 85 90 95 Tyr Cys Ala Arg Leu Thr Tyr Gly Phe Asp Tyr Trp
Gly Gln Gly Thr 100 105 110 Leu Val Thr Val Ser Ser Ala Ser Thr Lys
Gly Pro Ser Val Phe Pro 115 120 125 Leu Ala Pro Ser Ser Lys Ser Thr
Ser Gly Gly Thr Ala Ala Leu Gly 130 135 140 Cys Leu Val Lys Asp Tyr
Phe Pro Glu Pro Val Thr Val Ser Trp Asn 145 150 155 160 Ser Gly Ala
Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln 165 170 175 Ser
Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser 180 185
190 Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser
195 200 205 Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp
Lys Thr 210 215 220 His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala
Gly Gly Pro Ser 225 230 235 240 Val Phe Leu Phe Pro Pro Lys Pro Lys
Asp Thr Leu Met Ile Ser Arg 245 250 255 Thr Pro Glu Val Thr Cys Val
Val Val Asp Val Ser His Glu Asp Pro 260 265 270 Glu Val Lys Phe Asn
Trp Tyr Val Asp Gly Val Glu Val His Asn Ala 275 280 285 Lys Thr Lys
Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val 290 295 300 Ser
Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr 305 310
315 320 Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys
Thr 325 330 335 Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val
Tyr Thr Leu 340 345 350 Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln
Val Ser Leu Ser Cys 355 360 365 Ala Val Lys Gly Phe Tyr Pro Ser Asp
Ile Ala Val Glu Trp Glu Ser 370 375 380 Asn Gly Gln Pro Glu Asn Asn
Tyr Lys Thr Thr Pro Pro Val Leu Asp 385 390 395 400 Ser Asp Gly Ser
Phe Phe Leu Val Ser Lys Leu Thr Val Asp Lys Ser 405 410 415 Arg Trp
Gln Gln Gly Asn Val Phe Ser Cys Ser Val Leu His Glu Ala 420 425 430
Leu His Ser His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys 435
440 445 <210> SEQ ID NO 54 <211> LENGTH: 219
<212> TYPE: PRT <213> ORGANISM: Artificial Sequence
<220> FEATURE: <221> NAME/KEY: source <223> OTHER
INFORMATION: /note="Description of Artificial Sequence: Synthetic
polypeptide" <400> SEQUENCE: 54 Asp Ile Val Met Thr Gln Thr
Pro Leu Ser Leu Pro Val Thr Pro Gly 1 5 10 15 Glu Pro Ala Ser Ile
Ser Cys Gln Ser Ser Lys Ser Leu Leu His Ser 20 25 30 Thr Gly Lys
Thr Tyr Leu Asn Trp Tyr Leu Gln Lys Pro Gly Gln Ser 35 40 45 Pro
Gln Leu Leu Ile Tyr Trp Met Ser Thr Arg Ala Ser Gly Val Pro 50 55
60 Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Lys Ile
65 70 75 80 Ser Arg Val Glu Ala Glu Asp Val Gly Val Tyr Tyr Cys Gln
Gln Phe 85 90 95 Leu Glu Tyr Pro Phe Thr Phe Gly Gln Gly Thr Lys
Val Glu Ile Lys 100 105 110 Arg Thr Val Ala Ala Pro Ser Val Phe Ile
Phe Pro Pro Ser Asp Glu 115 120 125 Gln Leu Lys Ser Gly Thr Ala Ser
Val Val Cys Leu Leu Asn Asn Phe 130 135 140 Tyr Pro Arg Glu Ala Lys
Val Gln Trp Lys Val Asp Asn Ala Leu Gln 145 150 155 160 Ser Gly Asn
Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser 165 170 175 Thr
Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu 180 185
190 Lys His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser
195 200 205 Pro Val Thr Lys Ser Phe Asn Arg Gly Glu Cys 210 215
<210> SEQ ID NO 55 <211> LENGTH: 181 <212> TYPE:
PRT <213> ORGANISM: Homo sapiens <400> SEQUENCE: 55 Asn
Ser Val Ile Ile Val Asp Lys Asn Gly Arg Leu Val Tyr Leu Val 1 5 10
15 Glu Asn Pro Gly Gly Tyr Val Ala Tyr Ser Lys Ala Ala Thr Val Thr
20 25 30 Gly Lys Leu Val His Ala Asn Phe Gly Thr Lys Lys Asp Phe
Glu Asp 35 40 45 Leu Tyr Thr Pro Val Asn Gly Ser Ile Val Ile Val
Arg Ala Gly Lys 50 55 60 Ile Thr Phe Ala Glu Lys Val Ala Asn Ala
Glu Ser Leu Asn Ala Ile 65 70 75 80 Gly Val Leu Ile Tyr Met Asp Gln
Thr Lys Phe Pro Ile Val Asn Ala 85 90 95 Glu Leu Ser Phe Phe Gly
His Ala His Leu Gly Thr Gly Asp Pro Tyr 100 105 110 Thr Pro Gly Phe
Pro Ser Phe Asn His Thr Gln Phe Pro Pro Ser Arg 115 120 125 Ser Ser
Gly Leu Pro Asn Ile Pro Val Gln Thr Ile Ser Arg Ala Ala 130 135 140
Ala Glu Lys Leu Phe Gly Asn Met Glu Gly Asp Cys Pro Ser Asp Trp 145
150 155 160 Lys Thr Asp Ser Thr Cys Arg Met Val Thr Ser Glu Ser Lys
Asn Val 165 170 175 Lys Leu Thr Val Ser 180 <210> SEQ ID NO
56 <211> LENGTH: 10 <212> TYPE: PRT <213>
ORGANISM: Artificial Sequence <220> FEATURE: <221>
NAME/KEY: source <223> OTHER INFORMATION: /note="Description
of Artificial Sequence: Synthetic peptide" <400> SEQUENCE: 56
Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser 1 5 10 <210> SEQ ID
NO 57 <211> LENGTH: 10 <212> TYPE: PRT <213>
ORGANISM: Homo sapiens <400> SEQUENCE: 57 Asp Lys Thr His Thr
Cys Pro Pro Cys Pro 1 5 10 <210> SEQ ID NO 58 <211>
LENGTH: 7 <212> TYPE: PRT <213> ORGANISM: Artificial
Sequence <220> FEATURE: <221> NAME/KEY: source
<223> OTHER INFORMATION: /note="Description of Artificial
Sequence: Synthetic peptide" <220> FEATURE: <221>
NAME/KEY: MOD_RES <222> LOCATION: (2)..(2) <223> OTHER
INFORMATION: Any amino acid <400> SEQUENCE: 58 Tyr Xaa Thr
Glu Trp Ser Ser 1 5 <210> SEQ ID NO 59 <211> LENGTH: 9
<212> TYPE: PRT <213> ORGANISM: Artificial Sequence
<220> FEATURE: <221> NAME/KEY: source <223> OTHER
INFORMATION: /note="Description of Artificial Sequence: Synthetic
peptide" <220> FEATURE: <221> NAME/KEY: MOD_RES
<222> LOCATION: (2)..(3) <223> OTHER INFORMATION: Any
amino acid <220> FEATURE: <221> NAME/KEY: MOD_RES
<222> LOCATION: (5)..(8) <223> OTHER INFORMATION: Any
amino acid <400> SEQUENCE: 59 Thr Xaa Xaa Glu Xaa Xaa Xaa Xaa
Phe 1 5 <210> SEQ ID NO 60 <211> LENGTH: 760
<212> TYPE: PRT <213> ORGANISM: Macaca fascicularis
<400> SEQUENCE: 60 Met Met Asp Gln Ala Arg Ser Ala Phe Ser
Asn Leu Phe Gly Gly Glu 1 5 10 15 Pro Leu Ser Tyr Thr Arg Phe Ser
Leu Ala Arg Gln Val Asp Gly Asp 20 25 30
Asn Ser His Val Glu Met Lys Leu Gly Val Asp Glu Glu Glu Asn Thr 35
40 45 Asp Asn Asn Thr Lys Ala Asn Gly Thr Lys Pro Lys Arg Cys Gly
Gly 50 55 60 Asn Ile Cys Tyr Gly Thr Ile Ala Val Ile Ile Phe Phe
Leu Ile Gly 65 70 75 80 Phe Met Ile Gly Tyr Leu Gly Tyr Cys Lys Gly
Val Glu Pro Lys Thr 85 90 95 Glu Cys Glu Arg Leu Ala Gly Thr Glu
Ser Pro Ala Arg Glu Glu Pro 100 105 110 Glu Glu Asp Phe Pro Ala Ala
Pro Arg Leu Tyr Trp Asp Asp Leu Lys 115 120 125 Arg Lys Leu Ser Glu
Lys Leu Asp Thr Thr Asp Phe Thr Ser Thr Ile 130 135 140 Lys Leu Leu
Asn Glu Asn Leu Tyr Val Pro Arg Glu Ala Gly Ser Gln 145 150 155 160
Lys Asp Glu Asn Leu Ala Leu Tyr Ile Glu Asn Gln Phe Arg Glu Phe 165
170 175 Lys Leu Ser Lys Val Trp Arg Asp Gln His Phe Val Lys Ile Gln
Val 180 185 190 Lys Asp Ser Ala Gln Asn Ser Val Ile Ile Val Asp Lys
Asn Gly Gly 195 200 205 Leu Val Tyr Leu Val Glu Asn Pro Gly Gly Tyr
Val Ala Tyr Ser Lys 210 215 220 Ala Ala Thr Val Thr Gly Lys Leu Val
His Ala Asn Phe Gly Thr Lys 225 230 235 240 Lys Asp Phe Glu Asp Leu
Asp Ser Pro Val Asn Gly Ser Ile Val Ile 245 250 255 Val Arg Ala Gly
Lys Ile Thr Phe Ala Glu Lys Val Ala Asn Ala Glu 260 265 270 Ser Leu
Asn Ala Ile Gly Val Leu Ile Tyr Met Asp Gln Thr Lys Phe 275 280 285
Pro Ile Val Lys Ala Asp Leu Ser Phe Phe Gly His Ala His Leu Gly 290
295 300 Thr Gly Asp Pro Tyr Thr Pro Gly Phe Pro Ser Phe Asn His Thr
Gln 305 310 315 320 Phe Pro Pro Ser Gln Ser Ser Gly Leu Pro Asn Ile
Pro Val Gln Thr 325 330 335 Ile Ser Arg Ala Ala Ala Glu Lys Leu Phe
Gly Asn Met Glu Gly Asp 340 345 350 Cys Pro Ser Asp Trp Lys Thr Asp
Ser Thr Cys Lys Met Val Thr Ser 355 360 365 Glu Asn Lys Ser Val Lys
Leu Thr Val Ser Asn Val Leu Lys Glu Thr 370 375 380 Lys Ile Leu Asn
Ile Phe Gly Val Ile Lys Gly Phe Val Glu Pro Asp 385 390 395 400 His
Tyr Val Val Val Gly Ala Gln Arg Asp Ala Trp Gly Pro Gly Ala 405 410
415 Ala Lys Ser Ser Val Gly Thr Ala Leu Leu Leu Lys Leu Ala Gln Met
420 425 430 Phe Ser Asp Met Val Leu Lys Asp Gly Phe Gln Pro Ser Arg
Ser Ile 435 440 445 Ile Phe Ala Ser Trp Ser Ala Gly Asp Phe Gly Ser
Val Gly Ala Thr 450 455 460 Glu Trp Leu Glu Gly Tyr Leu Ser Ser Leu
His Leu Lys Ala Phe Thr 465 470 475 480 Tyr Ile Asn Leu Asp Lys Ala
Val Leu Gly Thr Ser Asn Phe Lys Val 485 490 495 Ser Ala Ser Pro Leu
Leu Tyr Thr Leu Ile Glu Lys Thr Met Gln Asp 500 505 510 Val Lys His
Pro Val Thr Gly Arg Ser Leu Tyr Gln Asp Ser Asn Trp 515 520 525 Ala
Ser Lys Val Glu Lys Leu Thr Leu Asp Asn Ala Ala Phe Pro Phe 530 535
540 Leu Ala Tyr Ser Gly Ile Pro Ala Val Ser Phe Cys Phe Cys Glu Asp
545 550 555 560 Thr Asp Tyr Pro Tyr Leu Gly Thr Thr Met Asp Thr Tyr
Lys Glu Leu 565 570 575 Val Glu Arg Ile Pro Glu Leu Asn Lys Val Ala
Arg Ala Ala Ala Glu 580 585 590 Val Ala Gly Gln Phe Val Ile Lys Leu
Thr His Asp Thr Glu Leu Asn 595 600 605 Leu Asp Tyr Glu Arg Tyr Asn
Ser Gln Leu Leu Leu Phe Leu Arg Asp 610 615 620 Leu Asn Gln Tyr Arg
Ala Asp Val Lys Glu Met Gly Leu Ser Leu Gln 625 630 635 640 Trp Leu
Tyr Ser Ala Arg Gly Asp Phe Phe Arg Ala Thr Ser Arg Leu 645 650 655
Thr Thr Asp Phe Arg Asn Ala Glu Lys Arg Asp Lys Phe Val Met Lys 660
665 670 Lys Leu Asn Asp Arg Val Met Arg Val Glu Tyr Tyr Phe Leu Ser
Pro 675 680 685 Tyr Val Ser Pro Lys Glu Ser Pro Phe Arg His Val Phe
Trp Gly Ser 690 695 700 Gly Ser His Thr Leu Ser Ala Leu Leu Glu Ser
Leu Lys Leu Arg Arg 705 710 715 720 Gln Asn Asn Ser Ala Phe Asn Glu
Thr Leu Phe Arg Asn Gln Leu Ala 725 730 735 Leu Ala Thr Trp Thr Ile
Gln Gly Ala Ala Asn Ala Leu Ser Gly Asp 740 745 750 Val Trp Asp Ile
Asp Asn Glu Phe 755 760 <210> SEQ ID NO 61 <211>
LENGTH: 217 <212> TYPE: PRT <213> ORGANISM: Artificial
Sequence <220> FEATURE: <221> NAME/KEY: source
<223> OTHER INFORMATION: /note="Description of Artificial
Sequence: Synthetic polypeptide" <400> SEQUENCE: 61 Ala Pro
Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys 1 5 10 15
Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val 20
25 30 Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp
Tyr 35 40 45 Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro
Arg Glu Glu 50 55 60 Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val
Leu Thr Val Leu His 65 70 75 80 Gln Asp Trp Leu Asn Gly Lys Glu Tyr
Lys Cys Lys Val Ser Asn Lys 85 90 95 Ala Leu Pro Ala Pro Ile Glu
Lys Thr Ile Ser Lys Ala Lys Gly Gln 100 105 110 Pro Arg Glu Pro Gln
Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu 115 120 125 Thr Lys Asn
Gln Val Ser Leu Ser Cys Ala Val Lys Gly Phe Tyr Pro 130 135 140 Ser
Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn 145 150
155 160 Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe
Leu 165 170 175 Val Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln
Gly Asn Val 180 185 190 Phe Ser Cys Ser Val Leu His Glu Ala Leu His
Ser His Tyr Thr Gln 195 200 205 Lys Ser Leu Ser Leu Ser Pro Gly Lys
210 215 <210> SEQ ID NO 62 <211> LENGTH: 760
<212> TYPE: PRT <213> ORGANISM: Homo sapiens
<400> SEQUENCE: 62 Met Met Asp Gln Ala Arg Ser Ala Phe Ser
Asn Leu Phe Gly Gly Glu 1 5 10 15 Pro Leu Ser Tyr Thr Arg Phe Ser
Leu Ala Arg Gln Val Asp Gly Asp 20 25 30 Asn Ser His Val Glu Met
Lys Leu Ala Val Asp Glu Glu Glu Asn Ala 35 40 45 Asp Asn Asn Thr
Lys Ala Asn Val Thr Lys Pro Lys Arg Cys Ser Gly 50 55 60 Ser Ile
Cys Tyr Gly Thr Ile Ala Val Ile Val Phe Phe Leu Ile Gly 65 70 75 80
Phe Met Ile Gly Tyr Leu Gly Tyr Cys Lys Gly Val Glu Pro Lys Thr 85
90 95 Glu Cys Glu Arg Leu Ala Gly Thr Glu Ser Pro Val Arg Glu Glu
Pro 100 105 110 Gly Glu Asp Phe Pro Ala Ala Arg Arg Leu Tyr Trp Asp
Asp Leu Lys 115 120 125 Arg Lys Leu Ser Glu Lys Leu Asp Ser Thr Asp
Phe Thr Gly Thr Ile 130 135 140 Lys Leu Leu Asn Glu Asn Ser Tyr Val
Pro Arg Glu Ala Gly Ser Gln 145 150 155 160 Lys Asp Glu Asn Leu Ala
Leu Tyr Val Glu Asn Gln Phe Arg Glu Phe 165 170 175 Lys Leu Ser Lys
Val Trp Arg Asp Gln His Phe Val Lys Ile Gln Val 180 185 190 Lys Asp
Ser Ala Gln Asn Ser Val Ile Ile Val Asp Lys Asn Gly Arg 195 200 205
Leu Val Tyr Leu Val Glu Asn Pro Gly Gly Tyr Val Ala Tyr Ser Lys 210
215 220 Ala Ala Thr Val Thr Gly Lys Leu Val His Ala Asn Phe Gly Thr
Lys 225 230 235 240 Lys Asp Phe Glu Asp Leu Tyr Thr Pro Val Asn Gly
Ser Ile Val Ile 245 250 255 Val Arg Ala Gly Lys Ile Thr Phe Ala Glu
Lys Val Ala Asn Ala Glu 260 265 270
Ser Leu Asn Ala Ile Gly Val Leu Ile Tyr Met Asp Gln Thr Lys Phe 275
280 285 Pro Ile Val Asn Ala Glu Leu Ser Phe Phe Gly His Ala His Leu
Gly 290 295 300 Thr Gly Asp Pro Tyr Thr Pro Gly Phe Pro Ser Phe Asn
His Thr Gln 305 310 315 320 Phe Pro Pro Ser Arg Ser Ser Gly Leu Pro
Asn Ile Pro Val Gln Thr 325 330 335 Ile Ser Arg Ala Ala Ala Glu Lys
Leu Phe Gly Asn Met Glu Gly Asp 340 345 350 Cys Pro Ser Asp Trp Lys
Thr Asp Ser Thr Cys Arg Met Val Thr Ser 355 360 365 Glu Ser Lys Asn
Val Lys Leu Thr Val Ser Asn Val Leu Lys Glu Ile 370 375 380 Lys Ile
Leu Asn Ile Phe Gly Val Ile Lys Gly Phe Val Glu Pro Asp 385 390 395
400 His Tyr Val Val Val Gly Ala Gln Arg Asp Ala Trp Gly Pro Gly Ala
405 410 415 Ala Lys Ser Gly Val Gly Thr Ala Leu Leu Leu Lys Leu Ala
Gln Met 420 425 430 Phe Ser Asp Met Val Leu Lys Asp Gly Phe Gln Pro
Ser Arg Ser Ile 435 440 445 Ile Phe Ala Ser Trp Ser Ala Gly Asp Phe
Gly Ser Val Gly Ala Thr 450 455 460 Glu Trp Leu Glu Gly Tyr Leu Ser
Ser Leu His Leu Lys Ala Phe Thr 465 470 475 480 Tyr Ile Asn Leu Asp
Lys Ala Val Leu Gly Thr Ser Asn Phe Lys Val 485 490 495 Ser Ala Ser
Pro Leu Leu Tyr Thr Leu Ile Glu Lys Thr Met Gln Asn 500 505 510 Val
Lys His Pro Val Thr Gly Gln Phe Leu Tyr Gln Asp Ser Asn Trp 515 520
525 Ala Ser Lys Val Glu Lys Leu Thr Leu Asp Asn Ala Ala Phe Pro Phe
530 535 540 Leu Ala Tyr Ser Gly Ile Pro Ala Val Ser Phe Cys Phe Cys
Glu Asp 545 550 555 560 Thr Asp Tyr Pro Tyr Leu Gly Thr Thr Met Asp
Thr Tyr Lys Glu Leu 565 570 575 Ile Glu Arg Ile Pro Glu Leu Asn Lys
Val Ala Arg Ala Ala Ala Glu 580 585 590 Val Ala Gly Gln Phe Val Ile
Lys Leu Thr His Asp Val Glu Leu Asn 595 600 605 Leu Asp Tyr Glu Arg
Tyr Asn Ser Gln Leu Leu Ser Phe Val Arg Asp 610 615 620 Leu Asn Gln
Tyr Arg Ala Asp Ile Lys Glu Met Gly Leu Ser Leu Gln 625 630 635 640
Trp Leu Tyr Ser Ala Arg Gly Asp Phe Phe Arg Ala Thr Ser Arg Leu 645
650 655 Thr Thr Asp Phe Gly Asn Ala Glu Lys Thr Asp Arg Phe Val Met
Lys 660 665 670 Lys Leu Asn Asp Arg Val Met Arg Val Glu Tyr His Phe
Leu Ser Pro 675 680 685 Tyr Val Ser Pro Lys Glu Ser Pro Phe Arg His
Val Phe Trp Gly Ser 690 695 700 Gly Ser His Thr Leu Pro Ala Leu Leu
Glu Asn Leu Lys Leu Arg Lys 705 710 715 720 Gln Asn Asn Gly Ala Phe
Asn Glu Thr Leu Phe Arg Asn Gln Leu Ala 725 730 735 Leu Ala Thr Trp
Thr Ile Gln Gly Ala Ala Asn Ala Leu Ser Gly Asp 740 745 750 Val Trp
Asp Ile Asp Asn Glu Phe 755 760 <210> SEQ ID NO 63
<211> LENGTH: 216 <212> TYPE: PRT <213> ORGANISM:
Artificial Sequence <220> FEATURE: <221> NAME/KEY:
source <223> OTHER INFORMATION: /note="Description of
Artificial Sequence: Synthetic polypeptide" <400> SEQUENCE:
63 Ala Pro Glu Ala Ala Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys
1 5 10 15 Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr
Cys Val 20 25 30 Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys
Phe Asn Trp Tyr 35 40 45 Val Asp Gly Val Glu Val His Asn Ala Lys
Thr Lys Pro Arg Glu Glu 50 55 60 Gln Tyr Asn Ser Thr Tyr Arg Val
Val Ser Val Leu Thr Val Leu His 65 70 75 80 Gln Asp Trp Leu Asn Gly
Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys 85 90 95 Ala Leu Pro Ala
Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln 100 105 110 Pro Arg
Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu 115 120 125
Thr Lys Asn Gln Val Ser Leu Ser Cys Ala Val Lys Gly Phe Tyr Pro 130
135 140 Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn
Asn 145 150 155 160 Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly
Ser Phe Phe Leu 165 170 175 Val Ser Lys Leu Thr Val Asp Lys Ser Arg
Trp Gln Gln Gly Asn Val 180 185 190 Phe Ser Cys Ser Val Leu His Glu
Ala Leu His Ser His Tyr Thr Gln 195 200 205 Lys Ser Leu Ser Leu Ser
Pro Gly 210 215 <210> SEQ ID NO 64 <211> LENGTH: 216
<212> TYPE: PRT <213> ORGANISM: Artificial Sequence
<220> FEATURE: <221> NAME/KEY: source <223> OTHER
INFORMATION: /note="Description of Artificial Sequence: Synthetic
polypeptide" <400> SEQUENCE: 64 Ala Pro Glu Ala Ala Gly Gly
Pro Ser Val Phe Leu Phe Pro Pro Lys 1 5 10 15 Pro Lys Asp Thr Leu
Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val 20 25 30 Val Val Asp
Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr 35 40 45 Val
Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu 50 55
60 Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His
65 70 75 80 Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser
Asn Lys 85 90 95 Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys
Ala Lys Gly Gln 100 105 110 Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro
Pro Ser Arg Asp Glu Leu 115 120 125 Thr Lys Asn Gln Val Ser Leu Trp
Cys Leu Val Lys Gly Phe Tyr Pro 130 135 140 Ser Asp Ile Ala Val Trp
Trp Glu Ser Tyr Gly Thr Glu Trp Ser Ser 145 150 155 160 Tyr Lys Thr
Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu 165 170 175 Tyr
Ser Lys Leu Thr Val Thr Lys Glu Glu Trp Gln Gln Gly Phe Val 180 185
190 Phe Ser Cys Ser Val Leu His Glu Ala Leu His Ser His Tyr Thr Gln
195 200 205 Lys Ser Leu Ser Leu Ser Pro Gly 210 215 <210> SEQ
ID NO 65 <211> LENGTH: 447 <212> TYPE: PRT <213>
ORGANISM: Artificial Sequence <220> FEATURE: <221>
NAME/KEY: source <223> OTHER INFORMATION: /note="Description
of Artificial Sequence: Synthetic polypeptide" <400>
SEQUENCE: 65 Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln
Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe
Thr Phe Thr Asp Phe 20 25 30 Tyr Met Ser Trp Val Arg Gln Ala Pro
Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Val Ile Arg Asn Lys Ala
Asn Ala Tyr Thr Ala Gly Tyr Asn Pro 50 55 60 Ser Val Lys Gly Arg
Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr 65 70 75 80 Leu Tyr Leu
Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr 85 90 95 Tyr
Cys Ala Arg Leu Thr Tyr Gly Phe Asp Tyr Trp Gly Gln Gly Thr 100 105
110 Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro
115 120 125 Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala
Leu Gly 130 135 140 Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr
Val Ser Trp Asn 145 150 155 160 Ser Gly Ala Leu Thr Ser Gly Val His
Thr Phe Pro Ala Val Leu Gln 165 170 175 Ser Ser Gly Leu Tyr Ser Leu
Ser Ser Val Val Thr Val Pro Ser Ser 180 185 190 Ser Leu Gly Thr Gln
Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser 195 200 205
Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr 210
215 220 His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Gly Pro
Ser 225 230 235 240 Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu
Met Ile Ser Arg 245 250 255 Thr Pro Glu Val Thr Cys Val Val Val Asp
Val Ser His Glu Asp Pro 260 265 270 Glu Val Lys Phe Asn Trp Tyr Val
Asp Gly Val Glu Val His Asn Ala 275 280 285 Lys Thr Lys Pro Arg Glu
Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val 290 295 300 Ser Val Leu Thr
Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr 305 310 315 320 Lys
Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr 325 330
335 Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu
340 345 350 Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu
Trp Cys 355 360 365 Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val
Trp Trp Glu Ser 370 375 380 Tyr Gly Thr Glu Trp Ser Ser Tyr Lys Thr
Thr Pro Pro Val Leu Asp 385 390 395 400 Ser Asp Gly Ser Phe Phe Leu
Tyr Ser Lys Leu Thr Val Thr Lys Glu 405 410 415 Glu Trp Gln Gln Gly
Phe Val Phe Ser Cys Ser Val Leu His Glu Ala 420 425 430 Leu His Ser
His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly 435 440 445
<210> SEQ ID NO 66 <211> LENGTH: 216 <212> TYPE:
PRT <213> ORGANISM: Artificial Sequence <220> FEATURE:
<221> NAME/KEY: source <223> OTHER INFORMATION:
/note="Description of Artificial Sequence: Synthetic polypeptide"
<400> SEQUENCE: 66 Ala Pro Glu Ala Ala Gly Gly Pro Ser Val
Phe Leu Phe Pro Pro Lys 1 5 10 15 Pro Lys Asp Thr Leu Met Ile Ser
Arg Thr Pro Glu Val Thr Cys Val 20 25 30 Val Val Asp Val Ser His
Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr 35 40 45 Val Asp Gly Val
Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu 50 55 60 Gln Tyr
Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His 65 70 75 80
Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys 85
90 95 Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly
Gln 100 105 110 Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg
Asp Glu Leu 115 120 125 Thr Lys Asn Gln Val Ser Leu Trp Cys Leu Val
Lys Gly Phe Tyr Pro 130 135 140 Ser Asp Ile Ala Val Glu Trp Glu Ser
Phe Gly Thr Glu Trp Ser Asn 145 150 155 160 Tyr Lys Thr Thr Pro Pro
Val Leu Asp Ser Asp Gly Ser Phe Phe Leu 165 170 175 Tyr Ser Lys Leu
Thr Val Ser Lys Glu Glu Trp Gln Gln Gly Phe Val 180 185 190 Phe Ser
Cys Ser Val Leu His Glu Ala Leu His Ser His Tyr Thr Gln 195 200 205
Lys Ser Leu Ser Leu Ser Pro Gly 210 215 <210> SEQ ID NO 67
<211> LENGTH: 447 <212> TYPE: PRT <213> ORGANISM:
Artificial Sequence <220> FEATURE: <221> NAME/KEY:
source <223> OTHER INFORMATION: /note="Description of
Artificial Sequence: Synthetic polypeptide" <400> SEQUENCE:
67 Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly
1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Thr
Asp Phe 20 25 30 Tyr Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly
Leu Glu Trp Val 35 40 45 Ser Val Ile Arg Asn Lys Ala Asn Ala Tyr
Thr Ala Gly Tyr Asn Pro 50 55 60 Ser Val Lys Gly Arg Phe Thr Ile
Ser Arg Asp Asn Ser Lys Asn Thr 65 70 75 80 Leu Tyr Leu Gln Met Asn
Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr 85 90 95 Tyr Cys Ala Arg
Leu Thr Tyr Gly Phe Asp Tyr Trp Gly Gln Gly Thr 100 105 110 Leu Val
Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro 115 120 125
Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly 130
135 140 Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp
Asn 145 150 155 160 Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro
Ala Val Leu Gln 165 170 175 Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val
Val Thr Val Pro Ser Ser 180 185 190 Ser Leu Gly Thr Gln Thr Tyr Ile
Cys Asn Val Asn His Lys Pro Ser 195 200 205 Asn Thr Lys Val Asp Lys
Lys Val Glu Pro Lys Ser Cys Asp Lys Thr 210 215 220 His Thr Cys Pro
Pro Cys Pro Ala Pro Glu Ala Ala Gly Gly Pro Ser 225 230 235 240 Val
Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg 245 250
255 Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro
260 265 270 Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His
Asn Ala 275 280 285 Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr
Tyr Arg Val Val 290 295 300 Ser Val Leu Thr Val Leu His Gln Asp Trp
Leu Asn Gly Lys Glu Tyr 305 310 315 320 Lys Cys Lys Val Ser Asn Lys
Ala Leu Pro Ala Pro Ile Glu Lys Thr 325 330 335 Ile Ser Lys Ala Lys
Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu 340 345 350 Pro Pro Ser
Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Trp Cys 355 360 365 Leu
Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser 370 375
380 Phe Gly Thr Glu Trp Ser Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp
385 390 395 400 Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val
Ser Lys Glu 405 410 415 Glu Trp Gln Gln Gly Phe Val Phe Ser Cys Ser
Val Leu His Glu Ala 420 425 430 Leu His Ser His Tyr Thr Gln Lys Ser
Leu Ser Leu Ser Pro Gly 435 440 445 <210> SEQ ID NO 68
<211> LENGTH: 216 <212> TYPE: PRT <213> ORGANISM:
Artificial Sequence <220> FEATURE: <221> NAME/KEY:
source <223> OTHER INFORMATION: /note="Description of
Artificial Sequence: Synthetic polypeptide" <400> SEQUENCE:
68 Ala Pro Glu Ala Ala Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys
1 5 10 15 Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr
Cys Val 20 25 30 Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys
Phe Asn Trp Tyr 35 40 45 Val Asp Gly Val Glu Val His Asn Ala Lys
Thr Lys Pro Arg Glu Glu 50 55 60 Gln Tyr Asn Ser Thr Tyr Arg Val
Val Ser Val Leu Thr Val Leu His 65 70 75 80 Gln Asp Trp Leu Asn Gly
Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys 85 90 95 Ala Leu Pro Ala
Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln 100 105 110 Pro Arg
Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu 115 120 125
Thr Lys Asn Gln Val Ser Leu Trp Cys Leu Val Lys Gly Phe Tyr Pro 130
135 140 Ser Asp Ile Ala Val Glu Trp Glu Ser Tyr Gly Thr Glu Trp Val
Asn 145 150 155 160 Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly
Ser Phe Phe Leu 165 170 175 Tyr Ser Lys Leu Thr Val Thr Lys Glu Glu
Trp Gln Gln Gly Phe Val 180 185 190 Phe Ser Cys Ser Val Leu His Glu
Ala Leu His Ser His Tyr Thr Gln 195 200 205 Lys Ser Leu Ser Leu Ser
Pro Gly 210 215
<210> SEQ ID NO 69 <211> LENGTH: 447 <212> TYPE:
PRT <213> ORGANISM: Artificial Sequence <220> FEATURE:
<221> NAME/KEY: source <223> OTHER INFORMATION:
/note="Description of Artificial Sequence: Synthetic polypeptide"
<400> SEQUENCE: 69 Glu Val Gln Leu Val Glu Ser Gly Gly Gly
Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala
Ser Gly Phe Thr Phe Thr Asp Phe 20 25 30 Tyr Met Ser Trp Val Arg
Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Val Ile Arg
Asn Lys Ala Asn Ala Tyr Thr Ala Gly Tyr Asn Pro 50 55 60 Ser Val
Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr 65 70 75 80
Leu Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr 85
90 95 Tyr Cys Ala Arg Leu Thr Tyr Gly Phe Asp Tyr Trp Gly Gln Gly
Thr 100 105 110 Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser
Val Phe Pro 115 120 125 Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly
Thr Ala Ala Leu Gly 130 135 140 Cys Leu Val Lys Asp Tyr Phe Pro Glu
Pro Val Thr Val Ser Trp Asn 145 150 155 160 Ser Gly Ala Leu Thr Ser
Gly Val His Thr Phe Pro Ala Val Leu Gln 165 170 175 Ser Ser Gly Leu
Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser 180 185 190 Ser Leu
Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser 195 200 205
Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr 210
215 220 His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Gly Pro
Ser 225 230 235 240 Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu
Met Ile Ser Arg 245 250 255 Thr Pro Glu Val Thr Cys Val Val Val Asp
Val Ser His Glu Asp Pro 260 265 270 Glu Val Lys Phe Asn Trp Tyr Val
Asp Gly Val Glu Val His Asn Ala 275 280 285 Lys Thr Lys Pro Arg Glu
Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val 290 295 300 Ser Val Leu Thr
Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr 305 310 315 320 Lys
Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr 325 330
335 Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu
340 345 350 Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu
Trp Cys 355 360 365 Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val
Glu Trp Glu Ser 370 375 380 Tyr Gly Thr Glu Trp Val Asn Tyr Lys Thr
Thr Pro Pro Val Leu Asp 385 390 395 400 Ser Asp Gly Ser Phe Phe Leu
Tyr Ser Lys Leu Thr Val Thr Lys Glu 405 410 415 Glu Trp Gln Gln Gly
Phe Val Phe Ser Cys Ser Val Leu His Glu Ala 420 425 430 Leu His Ser
His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly 435 440 445
<210> SEQ ID NO 70 <211> LENGTH: 216 <212> TYPE:
PRT <213> ORGANISM: Artificial Sequence <220> FEATURE:
<221> NAME/KEY: source <223> OTHER INFORMATION:
/note="Description of Artificial Sequence: Synthetic polypeptide"
<400> SEQUENCE: 70 Ala Pro Glu Ala Ala Gly Gly Pro Ser Val
Phe Leu Phe Pro Pro Lys 1 5 10 15 Pro Lys Asp Thr Leu Met Ile Ser
Arg Thr Pro Glu Val Thr Cys Val 20 25 30 Val Val Asp Val Ser His
Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr 35 40 45 Val Asp Gly Val
Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu 50 55 60 Gln Tyr
Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His 65 70 75 80
Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys 85
90 95 Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly
Gln 100 105 110 Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg
Asp Glu Leu 115 120 125 Thr Lys Asn Gln Val Ser Leu Trp Cys Leu Val
Lys Gly Phe Tyr Pro 130 135 140 Ser Asp Ile Ala Val Glu Trp Glu Ser
Tyr Gly Thr Glu Trp Ser Asn 145 150 155 160 Tyr Lys Thr Thr Pro Pro
Val Leu Asp Ser Asp Gly Ser Phe Phe Leu 165 170 175 Tyr Ser Lys Leu
Thr Val Ser Lys Glu Glu Trp Gln Gln Gly Phe Val 180 185 190 Phe Ser
Cys Ser Val Leu His Glu Ala Leu His Ser His Tyr Thr Gln 195 200 205
Lys Ser Leu Ser Leu Ser Pro Gly 210 215 <210> SEQ ID NO 71
<211> LENGTH: 447 <212> TYPE: PRT <213> ORGANISM:
Artificial Sequence <220> FEATURE: <221> NAME/KEY:
source <223> OTHER INFORMATION: /note="Description of
Artificial Sequence: Synthetic polypeptide" <400> SEQUENCE:
71 Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly
1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Thr
Asp Phe 20 25 30 Tyr Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly
Leu Glu Trp Val 35 40 45 Ser Val Ile Arg Asn Lys Ala Asn Ala Tyr
Thr Ala Gly Tyr Asn Pro 50 55 60 Ser Val Lys Gly Arg Phe Thr Ile
Ser Arg Asp Asn Ser Lys Asn Thr 65 70 75 80 Leu Tyr Leu Gln Met Asn
Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr 85 90 95 Tyr Cys Ala Arg
Leu Thr Tyr Gly Phe Asp Tyr Trp Gly Gln Gly Thr 100 105 110 Leu Val
Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro 115 120 125
Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly 130
135 140 Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp
Asn 145 150 155 160 Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro
Ala Val Leu Gln 165 170 175 Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val
Val Thr Val Pro Ser Ser 180 185 190 Ser Leu Gly Thr Gln Thr Tyr Ile
Cys Asn Val Asn His Lys Pro Ser 195 200 205 Asn Thr Lys Val Asp Lys
Lys Val Glu Pro Lys Ser Cys Asp Lys Thr 210 215 220 His Thr Cys Pro
Pro Cys Pro Ala Pro Glu Ala Ala Gly Gly Pro Ser 225 230 235 240 Val
Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg 245 250
255 Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro
260 265 270 Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His
Asn Ala 275 280 285 Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr
Tyr Arg Val Val 290 295 300 Ser Val Leu Thr Val Leu His Gln Asp Trp
Leu Asn Gly Lys Glu Tyr 305 310 315 320 Lys Cys Lys Val Ser Asn Lys
Ala Leu Pro Ala Pro Ile Glu Lys Thr 325 330 335 Ile Ser Lys Ala Lys
Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu 340 345 350 Pro Pro Ser
Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Trp Cys 355 360 365 Leu
Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser 370 375
380 Tyr Gly Thr Glu Trp Ser Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp
385 390 395 400 Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val
Ser Lys Glu 405 410 415 Glu Trp Gln Gln Gly Phe Val Phe Ser Cys Ser
Val Leu His Glu Ala 420 425 430 Leu His Ser His Tyr Thr Gln Lys Ser
Leu Ser Leu Ser Pro Gly 435 440 445 <210> SEQ ID NO 72
<211> LENGTH: 447 <212> TYPE: PRT <213> ORGANISM:
Artificial Sequence <220> FEATURE:
<221> NAME/KEY: source <223> OTHER INFORMATION:
/note="Description of Artificial Sequence: Synthetic polypeptide"
<400> SEQUENCE: 72 Glu Val Gln Leu Val Glu Ser Gly Gly Gly
Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala
Ser Gly Phe Thr Phe Thr Asp Phe 20 25 30 Tyr Met Ser Trp Val Arg
Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Val Ile Arg
Asn Lys Ala Asn Ala Tyr Thr Ala Gly Tyr Asn Pro 50 55 60 Ser Val
Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr 65 70 75 80
Leu Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr 85
90 95 Tyr Cys Ala Arg Leu Thr Tyr Gly Phe Asp Tyr Trp Gly Gln Gly
Thr 100 105 110 Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser
Val Phe Pro 115 120 125 Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly
Thr Ala Ala Leu Gly 130 135 140 Cys Leu Val Lys Asp Tyr Phe Pro Glu
Pro Val Thr Val Ser Trp Asn 145 150 155 160 Ser Gly Ala Leu Thr Ser
Gly Val His Thr Phe Pro Ala Val Leu Gln 165 170 175 Ser Ser Gly Leu
Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser 180 185 190 Ser Leu
Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser 195 200 205
Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr 210
215 220 His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro
Ser 225 230 235 240 Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu
Met Ile Ser Arg 245 250 255 Thr Pro Glu Val Thr Cys Val Val Val Asp
Val Ser His Glu Asp Pro 260 265 270 Glu Val Lys Phe Asn Trp Tyr Val
Asp Gly Val Glu Val His Asn Ala 275 280 285 Lys Thr Lys Pro Arg Glu
Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val 290 295 300 Ser Val Leu Thr
Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr 305 310 315 320 Lys
Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr 325 330
335 Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu
340 345 350 Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu
Ser Cys 355 360 365 Ala Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val
Glu Trp Glu Ser 370 375 380 Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr
Thr Pro Pro Val Leu Asp 385 390 395 400 Ser Asp Gly Ser Phe Phe Leu
Val Ser Lys Leu Thr Val Asp Lys Ser 405 410 415 Arg Trp Gln Gln Gly
Asn Val Phe Ser Cys Ser Val Leu His Glu Ala 420 425 430 Leu His Ser
His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly 435 440 445
<210> SEQ ID NO 73 <211> LENGTH: 447 <212> TYPE:
PRT <213> ORGANISM: Artificial Sequence <220> FEATURE:
<221> NAME/KEY: source <223> OTHER INFORMATION:
/note="Description of Artificial Sequence: Synthetic polypeptide"
<400> SEQUENCE: 73 Glu Val Gln Leu Val Glu Ser Gly Gly Gly
Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala
Ser Gly Phe Thr Phe Thr Asp Phe 20 25 30 Tyr Met Ser Trp Val Arg
Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Val Ile Arg
Asn Lys Ala Asn Ala Tyr Thr Ala Gly Tyr Asn Pro 50 55 60 Ser Val
Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr 65 70 75 80
Leu Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr 85
90 95 Tyr Cys Ala Arg Leu Thr Tyr Gly Phe Asp Tyr Trp Gly Gln Gly
Thr 100 105 110 Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser
Val Phe Pro 115 120 125 Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly
Thr Ala Ala Leu Gly 130 135 140 Cys Leu Val Lys Asp Tyr Phe Pro Glu
Pro Val Thr Val Ser Trp Asn 145 150 155 160 Ser Gly Ala Leu Thr Ser
Gly Val His Thr Phe Pro Ala Val Leu Gln 165 170 175 Ser Ser Gly Leu
Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser 180 185 190 Ser Leu
Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser 195 200 205
Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr 210
215 220 His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Gly Pro
Ser 225 230 235 240 Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu
Met Ile Ser Arg 245 250 255 Thr Pro Glu Val Thr Cys Val Val Val Asp
Val Ser His Glu Asp Pro 260 265 270 Glu Val Lys Phe Asn Trp Tyr Val
Asp Gly Val Glu Val His Asn Ala 275 280 285 Lys Thr Lys Pro Arg Glu
Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val 290 295 300 Ser Val Leu Thr
Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr 305 310 315 320 Lys
Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr 325 330
335 Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu
340 345 350 Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu
Ser Cys 355 360 365 Ala Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val
Glu Trp Glu Ser 370 375 380 Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr
Thr Pro Pro Val Leu Asp 385 390 395 400 Ser Asp Gly Ser Phe Phe Leu
Val Ser Lys Leu Thr Val Asp Lys Ser 405 410 415 Arg Trp Gln Gln Gly
Asn Val Phe Ser Cys Ser Val Leu His Glu Ala 420 425 430 Leu His Ser
His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly 435 440 445
<210> SEQ ID NO 74 <211> LENGTH: 453 <212> TYPE:
PRT <213> ORGANISM: Artificial Sequence <220> FEATURE:
<221> NAME/KEY: source <223> OTHER INFORMATION:
/note="Description of Artificial Sequence: Synthetic polypeptide"
<400> SEQUENCE: 74 Gln Val Gln Leu Val Gln Ser Gly Ala Glu
Val Lys Lys Pro Gly Ala 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala
Ser Gly Tyr Ala Phe Ser Ser Gln 20 25 30 Trp Met Asn Trp Val Arg
Gln Ala Pro Gly Gln Arg Leu Glu Trp Ile 35 40 45 Gly Arg Ile Tyr
Pro Gly Gly Gly Asp Thr Asn Tyr Ala Gly Lys Phe 50 55 60 Gln Gly
Arg Val Thr Ile Thr Ala Asp Thr Ser Ala Ser Thr Ala Tyr 65 70 75 80
Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85
90 95 Ala Arg Leu Leu Arg Asn Gln Pro Gly Glu Ser Tyr Ala Met Asp
Tyr 100 105 110 Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser Ala Ser
Thr Lys Gly 115 120 125 Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys
Ser Thr Ser Gly Gly 130 135 140 Thr Ala Ala Leu Gly Cys Leu Val Lys
Asp Tyr Phe Pro Glu Pro Val 145 150 155 160 Thr Val Ser Trp Asn Ser
Gly Ala Leu Thr Ser Gly Val His Thr Phe 165 170 175 Pro Ala Val Leu
Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val 180 185 190 Thr Val
Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val 195 200 205
Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys 210
215 220 Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu
Leu 225 230 235 240 Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys
Pro Lys Asp Thr 245 250 255 Leu Met Ile Ser Arg Thr Pro Glu Val Thr
Cys Val Val Val Asp Val 260 265 270 Ser His Glu Asp Pro Glu Val Lys
Phe Asn Trp Tyr Val Asp Gly Val 275 280 285
Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser 290
295 300 Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp
Leu 305 310 315 320 Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys
Ala Leu Pro Ala 325 330 335 Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys
Gly Gln Pro Arg Glu Pro 340 345 350 Gln Val Tyr Thr Leu Pro Pro Ser
Arg Asp Glu Leu Thr Lys Asn Gln 355 360 365 Val Ser Leu Thr Cys Leu
Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala 370 375 380 Val Glu Trp Glu
Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr 385 390 395 400 Pro
Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu 405 410
415 Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser
420 425 430 Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser
Leu Ser 435 440 445 Leu Ser Pro Gly Lys 450 <210> SEQ ID NO
75 <211> LENGTH: 219 <212> TYPE: PRT <213>
ORGANISM: Artificial Sequence <220> FEATURE: <221>
NAME/KEY: source <223> OTHER INFORMATION: /note="Description
of Artificial Sequence: Synthetic polypeptide" <400>
SEQUENCE: 75 Asp Val Val Met Thr Gln Ser Pro Asp Ser Leu Ala Val
Ser Leu Gly 1 5 10 15 Glu Arg Ala Thr Ile Asn Cys Arg Ser Ser Gln
Ser Leu Val His Ser 20 25 30 Asn Arg Tyr Thr Tyr Leu His Trp Tyr
Gln Gln Lys Pro Gly Gln Ser 35 40 45 Pro Lys Leu Leu Ile Tyr Lys
Val Ser Asn Arg Phe Ser Gly Val Pro 50 55 60 Asp Arg Phe Ser Gly
Ser Gly Ser Gly Thr Asp Phe Thr Leu Lys Ile 65 70 75 80 Ser Arg Val
Glu Ala Glu Asp Val Gly Val Tyr Tyr Cys Ser Gln Ser 85 90 95 Thr
Arg Val Pro Tyr Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys 100 105
110 Arg Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu
115 120 125 Gln Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn
Asn Phe 130 135 140 Tyr Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp
Asn Ala Leu Gln 145 150 155 160 Ser Gly Asn Ser Gln Glu Ser Val Thr
Glu Gln Asp Ser Lys Asp Ser 165 170 175 Thr Tyr Ser Leu Ser Ser Thr
Leu Thr Leu Ser Lys Ala Asp Tyr Glu 180 185 190 Lys His Lys Val Tyr
Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser 195 200 205 Pro Val Thr
Lys Ser Phe Asn Arg Gly Glu Cys 210 215 <210> SEQ ID NO 76
<211> LENGTH: 453 <212> TYPE: PRT <213> ORGANISM:
Artificial Sequence <220> FEATURE: <221> NAME/KEY:
source <223> OTHER INFORMATION: /note="Description of
Artificial Sequence: Synthetic polypeptide" <400> SEQUENCE:
76 Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala
1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Ala Phe Ser
Ser Gln 20 25 30 Trp Met Asn Trp Val Arg Gln Ala Pro Gly Gln Arg
Leu Glu Trp Ile 35 40 45 Gly Arg Ile Tyr Pro Gly Gly Gly Asp Thr
Asn Tyr Ala Gly Lys Phe 50 55 60 Gln Gly Arg Val Thr Ile Thr Ala
Asp Thr Ser Ala Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu
Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Leu Leu
Arg Asn Gln Pro Gly Glu Ser Tyr Ala Met Asp Tyr 100 105 110 Trp Gly
Gln Gly Thr Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly 115 120 125
Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly 130
135 140 Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro
Val 145 150 155 160 Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly
Val His Thr Phe 165 170 175 Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr
Ser Leu Ser Ser Val Val 180 185 190 Thr Val Pro Ser Ser Ser Leu Gly
Thr Gln Thr Tyr Ile Cys Asn Val 195 200 205 Asn His Lys Pro Ser Asn
Thr Lys Val Asp Lys Lys Val Glu Pro Lys 210 215 220 Ser Cys Asp Lys
Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu 225 230 235 240 Leu
Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr 245 250
255 Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val
260 265 270 Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp
Gly Val 275 280 285 Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu
Gln Tyr Asn Ser 290 295 300 Thr Tyr Arg Val Val Ser Val Leu Thr Val
Leu His Gln Asp Trp Leu 305 310 315 320 Asn Gly Lys Glu Tyr Lys Cys
Lys Val Ser Asn Lys Ala Leu Pro Ala 325 330 335 Ser Ile Glu Lys Thr
Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro 340 345 350 Gln Val Tyr
Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln 355 360 365 Val
Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala 370 375
380 Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr
385 390 395 400 Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr
Ser Lys Leu 405 410 415 Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn
Val Phe Ser Cys Ser 420 425 430 Val Met His Gly Ala Leu His Asn His
Tyr Thr Gln Lys Ser Leu Ser 435 440 445 Leu Ser Pro Gly Lys 450
<210> SEQ ID NO 77 <211> LENGTH: 445 <212> TYPE:
PRT <213> ORGANISM: Artificial Sequence <220> FEATURE:
<221> NAME/KEY: source <223> OTHER INFORMATION:
/note="Description of Artificial Sequence: Synthetic polypeptide"
<400> SEQUENCE: 77 Gln Val Gln Leu Gln Glu Ser Gly Pro Gly
Leu Val Lys Pro Ser Glu 1 5 10 15 Thr Leu Ser Leu Thr Cys Ala Val
Ser Gly Tyr Ser Ile Thr Ser Asp 20 25 30 Tyr Ala Trp Asn Trp Ile
Arg Gln Pro Pro Gly Lys Gly Leu Glu Trp 35 40 45 Ile Gly Tyr Met
Ser Tyr Ser Gly Ser Thr Arg Tyr Asn Pro Ser Leu 50 55 60 Arg Ser
Arg Val Thr Ile Ser Val Asp Thr Ser Lys Asn Gln Phe Ser 65 70 75 80
Leu Lys Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr Cys 85
90 95 Ala Arg Gly Trp Pro Leu Ala Tyr Trp Gly Gln Gly Thr Leu Val
Thr 100 105 110 Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro
Leu Ala Pro 115 120 125 Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala
Leu Gly Cys Leu Val 130 135 140 Lys Asp Tyr Phe Pro Glu Pro Val Thr
Val Ser Trp Asn Ser Gly Ala 145 150 155 160 Leu Thr Ser Gly Val His
Thr Phe Pro Ala Val Leu Gln Ser Ser Gly 165 170 175 Leu Tyr Ser Leu
Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly 180 185 190 Thr Gln
Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys 195 200 205
Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys 210
215 220 Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe
Leu 225 230 235 240 Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser
Arg Thr Pro Glu 245 250 255 Val Thr Cys Val Val Val Asp Val Ser His
Glu Asp Pro Glu Val Lys 260 265 270 Phe Asn Trp Tyr Val Asp Gly Val
Glu Val His Asn Ala Lys Thr Lys
275 280 285 Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser
Val Leu 290 295 300 Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu
Tyr Lys Cys Lys 305 310 315 320 Val Ser Asn Lys Ala Leu Pro Ala Pro
Ile Glu Lys Thr Ile Ser Lys 325 330 335 Ala Lys Gly Gln Pro Arg Glu
Pro Gln Val Tyr Thr Leu Pro Pro Ser 340 345 350 Arg Asp Glu Leu Thr
Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys 355 360 365 Gly Phe Tyr
Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln 370 375 380 Pro
Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly 385 390
395 400 Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp
Gln 405 410 415 Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala
Leu His Asn 420 425 430 His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro
Gly Lys 435 440 445 <210> SEQ ID NO 78 <211> LENGTH:
214 <212> TYPE: PRT <213> ORGANISM: Artificial Sequence
<220> FEATURE: <221> NAME/KEY: source <223> OTHER
INFORMATION: /note="Description of Artificial Sequence: Synthetic
polypeptide" <400> SEQUENCE: 78 Glu Ile Val Met Thr Gln Ser
Pro Ala Thr Leu Ser Leu Ser Pro Gly 1 5 10 15 Glu Arg Ala Thr Leu
Ser Cys Ser Ala Ser Ser Ser Val Ser Tyr Met 20 25 30 Tyr Trp Tyr
Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu Ile Tyr 35 40 45 Asp
Thr Ser Asn Leu Ala Ser Gly Ile Pro Ala Arg Phe Ser Gly Ser 50 55
60 Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro Glu
65 70 75 80 Asp Phe Ala Val Tyr Tyr Cys Gln Gln Trp Ser Ser Tyr Pro
Pro Ile 85 90 95 Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys Arg
Thr Val Ala Ala 100 105 110 Pro Ser Val Phe Ile Phe Pro Pro Ser Asp
Glu Gln Leu Lys Ser Gly 115 120 125 Thr Ala Ser Val Val Cys Leu Leu
Asn Asn Phe Tyr Pro Arg Glu Ala 130 135 140 Lys Val Gln Trp Lys Val
Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln 145 150 155 160 Glu Ser Val
Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser 165 170 175 Ser
Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr 180 185
190 Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser
195 200 205 Phe Asn Arg Gly Glu Cys 210 <210> SEQ ID NO 79
<211> LENGTH: 450 <212> TYPE: PRT <213> ORGANISM:
Artificial Sequence <220> FEATURE: <221> NAME/KEY:
source <223> OTHER INFORMATION: /note="Description of
Artificial Sequence: Synthetic polypeptide" <400> SEQUENCE:
79 Gln Val Thr Leu Arg 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
Thr Ser 20 25 30 Gly Met Ser Val Gly Trp Ile Arg Gln Pro Pro Gly
Lys Ala Leu Glu 35 40 45 Trp Leu Ala Asp Ile Trp Trp Asp Asp Lys
Lys Asp Tyr Asn Pro Ser 50 55 60 Leu Lys Ser Arg Leu Thr Ile Ser
Lys Asp Thr Ser Lys Asn Gln Val 65 70 75 80 Val Leu Lys Val Thr Asn
Met Glu Pro Ala Asp Thr Ala Thr Tyr Tyr 85 90 95 Cys Ala Arg Ser
Met Ile Thr Asn Trp Tyr Phe Asp Val Trp Gly Ala 100 105 110 Gly Thr
Thr Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val 115 120 125
Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala 130
135 140 Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val
Ser 145 150 155 160 Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr
Phe Pro Ala Val 165 170 175 Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser
Ser Val Val Thr Val Pro 180 185 190 Ser Ser Ser Leu Gly Thr Gln Thr
Tyr Ile Cys Asn Val Asn His Lys 195 200 205 Pro Ser Asn Thr Lys Val
Asp Lys Lys Val Glu Pro Lys Ser Cys Asp 210 215 220 Lys Thr His Thr
Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Gly 225 230 235 240 Pro
Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile 245 250
255 Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu
260 265 270 Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu
Val His 275 280 285 Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn
Ser Thr Tyr Arg 290 295 300 Val Val Ser Val Leu Thr Val Leu His Gln
Asp Trp Leu Asn Gly Lys 305 310 315 320 Glu Tyr Lys Cys Lys Val Ser
Asn Lys Ala Leu Pro Ala Pro Ile Glu 325 330 335 Lys Thr Ile Ser Lys
Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr 340 345 350 Thr Leu Pro
Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu 355 360 365 Trp
Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Trp Trp 370 375
380 Glu Ser Tyr Gly Thr Glu Trp Ser Ser Tyr Lys Thr Thr Pro Pro Val
385 390 395 400 Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu
Thr Val Thr 405 410 415 Lys Glu Glu Trp Gln Gln Gly Phe Val Phe Ser
Cys Ser Val Met His 420 425 430 Glu Ala Leu His Asn His Tyr Thr Gln
Lys Ser Leu Ser Leu Ser Pro 435 440 445 Gly Lys 450 <210> SEQ
ID NO 80 <211> LENGTH: 450 <212> TYPE: PRT <213>
ORGANISM: Artificial Sequence <220> FEATURE: <221>
NAME/KEY: source <223> OTHER INFORMATION: /note="Description
of Artificial Sequence: Synthetic polypeptide" <400>
SEQUENCE: 80 Gln Val Thr Leu Arg 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 Thr Ser 20 25 30 Gly Met Ser Val Gly Trp Ile Arg Gln
Pro Pro Gly Lys Ala Leu Glu 35 40 45 Trp Leu Ala Asp Ile Trp Trp
Asp Asp Lys Lys Asp Tyr Asn Pro Ser 50 55 60 Leu Lys Ser Arg Leu
Thr Ile Ser Lys Asp Thr Ser Lys Asn Gln Val 65 70 75 80 Val Leu Lys
Val Thr Asn Met Glu Pro Ala Asp Thr Ala Thr Tyr Tyr 85 90 95 Cys
Ala Arg Ser Met Ile Thr Asn Trp Tyr Phe Asp Val Trp Gly Ala 100 105
110 Gly Thr Thr Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val
115 120 125 Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr
Ala Ala 130 135 140 Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro
Val Thr Val Ser 145 150 155 160 Trp Asn Ser Gly Ala Leu Thr Ser Gly
Val His Thr Phe Pro Ala Val 165 170 175 Leu Gln Ser Ser Gly Leu Tyr
Ser Leu Ser Ser Val Val Thr Val Pro 180 185 190 Ser Ser Ser Leu Gly
Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys 195 200 205 Pro Ser Asn
Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp 210 215 220 Lys
Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Gly 225 230
235 240 Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met
Ile 245 250 255 Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val
Ser His Glu 260 265 270 Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp
Gly Val Glu Val His 275 280 285
Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg 290
295 300 Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly
Lys 305 310 315 320 Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro
Ala Pro Ile Glu 325 330 335 Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro
Arg Glu Pro Gln Val Tyr 340 345 350 Thr Leu Pro Pro Ser Arg Asp Glu
Leu Thr Lys Asn Gln Val Ser Leu 355 360 365 Trp Cys Leu Val Lys Gly
Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp 370 375 380 Glu Ser Tyr Gly
Thr Glu Trp Val Asn Tyr Lys Thr Thr Pro Pro Val 385 390 395 400 Leu
Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Thr 405 410
415 Lys Glu Glu Trp Gln Gln Gly Phe Val Phe Ser Cys Ser Val Met His
420 425 430 Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu
Ser Pro 435 440 445 Gly Lys 450 <210> SEQ ID NO 81
<211> LENGTH: 450 <212> TYPE: PRT <213> ORGANISM:
Artificial Sequence <220> FEATURE: <221> NAME/KEY:
source <223> OTHER INFORMATION: /note="Description of
Artificial Sequence: Synthetic polypeptide" <400> SEQUENCE:
81 Gln Val Thr Leu Arg 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
Thr Ser 20 25 30 Gly Met Ser Val Gly Trp Ile Arg Gln Pro Pro Gly
Lys Ala Leu Glu 35 40 45 Trp Leu Ala Asp Ile Trp Trp Asp Asp Lys
Lys Asp Tyr Asn Pro Ser 50 55 60 Leu Lys Ser Arg Leu Thr Ile Ser
Lys Asp Thr Ser Lys Asn Gln Val 65 70 75 80 Val Leu Lys Val Thr Asn
Met Glu Pro Ala Asp Thr Ala Thr Tyr Tyr 85 90 95 Cys Ala Arg Ser
Met Ile Thr Asn Trp Tyr Phe Asp Val Trp Gly Ala 100 105 110 Gly Thr
Thr Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val 115 120 125
Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala 130
135 140 Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val
Ser 145 150 155 160 Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr
Phe Pro Ala Val 165 170 175 Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser
Ser Val Val Thr Val Pro 180 185 190 Ser Ser Ser Leu Gly Thr Gln Thr
Tyr Ile Cys Asn Val Asn His Lys 195 200 205 Pro Ser Asn Thr Lys Val
Asp Lys Lys Val Glu Pro Lys Ser Cys Asp 210 215 220 Lys Thr His Thr
Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Gly 225 230 235 240 Pro
Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile 245 250
255 Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu
260 265 270 Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu
Val His 275 280 285 Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn
Ser Thr Tyr Arg 290 295 300 Val Val Ser Val Leu Thr Val Leu His Gln
Asp Trp Leu Asn Gly Lys 305 310 315 320 Glu Tyr Lys Cys Lys Val Ser
Asn Lys Ala Leu Pro Ala Pro Ile Glu 325 330 335 Lys Thr Ile Ser Lys
Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr 340 345 350 Thr Leu Pro
Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu 355 360 365 Ser
Cys Ala Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp 370 375
380 Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val
385 390 395 400 Leu Asp Ser Asp Gly Ser Phe Phe Leu Val Ser Lys Leu
Thr Val Asp 405 410 415 Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser
Cys Ser Val Met His 420 425 430 Glu Ala Leu His Asn His Tyr Thr Gln
Lys Ser Leu Ser Leu Ser Pro 435 440 445 Gly Lys 450 <210> SEQ
ID NO 82 <211> LENGTH: 213 <212> TYPE: PRT <213>
ORGANISM: Artificial Sequence <220> FEATURE: <221>
NAME/KEY: source <223> OTHER INFORMATION: /note="Description
of Artificial Sequence: Synthetic polypeptide" <400>
SEQUENCE: 82 Asp Ile Gln Met Thr Gln Ser Pro Ser Thr Leu Ser Ala
Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Lys Cys Gln Leu
Ser Val Gly Tyr Met 20 25 30 His Trp Tyr Gln Gln Lys Pro Gly Lys
Ala Pro Lys Leu Leu Ile Tyr 35 40 45 Asp Thr Ser Lys Leu Ala Ser
Gly Val Pro Ser Arg Phe Ser Gly Ser 50 55 60 Gly Ser Gly Thr Glu
Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro Asp 65 70 75 80 Asp Phe Ala
Thr Tyr Tyr Cys Phe Gln Gly Ser Gly Tyr Pro Phe Thr 85 90 95 Phe
Gly Gly Gly Thr Lys Leu Glu Ile Lys Arg Thr Val Ala Ala Pro 100 105
110 Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly Thr
115 120 125 Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu
Ala Lys 130 135 140 Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly
Asn Ser Gln Glu 145 150 155 160 Ser Val Thr Glu Gln Asp Ser Lys Asp
Ser Thr Tyr Ser Leu Ser Ser 165 170 175 Thr Leu Thr Leu Ser Lys Ala
Asp Tyr Glu Lys His Lys Val Tyr Ala 180 185 190 Cys Glu Val Thr His
Gln Gly Leu Ser Ser Pro Val Thr Lys Ser Phe 195 200 205 Asn Arg Gly
Glu Cys 210 <210> SEQ ID NO 83 <211> LENGTH: 448
<212> TYPE: PRT <213> ORGANISM: Artificial Sequence
<220> FEATURE: <221> NAME/KEY: source <223> OTHER
INFORMATION: /note="Description of Artificial Sequence: Synthetic
polypeptide" <400> SEQUENCE: 83 Glu Val Gln Leu Val Glu Ser
Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser
Cys Ala Ala Ser Gly Phe Thr Phe Thr Asp Phe 20 25 30 Tyr Met Ser
Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser
Val Ile Arg Asn Lys Ala Asn Ala Tyr Thr Ala Gly Tyr Asn Pro 50 55
60 Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr
65 70 75 80 Leu Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala
Val Tyr 85 90 95 Tyr Cys Ala Arg Leu Thr Tyr Gly Phe Asp Tyr Trp
Gly Gln Gly Thr 100 105 110 Leu Val Thr Val Ser Ser Ala Ser Thr Lys
Gly Pro Ser Val Phe Pro 115 120 125 Leu Ala Pro Ser Ser Lys Ser Thr
Ser Gly Gly Thr Ala Ala Leu Gly 130 135 140 Cys Leu Val Lys Asp Tyr
Phe Pro Glu Pro Val Thr Val Ser Trp Asn 145 150 155 160 Ser Gly Ala
Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln 165 170 175 Ser
Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser 180 185
190 Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser
195 200 205 Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp
Lys Thr 210 215 220 His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala
Gly Gly Pro Ser 225 230 235 240 Val Phe Leu Phe Pro Pro Lys Pro Lys
Asp Thr Leu Met Ile Ser Arg 245 250 255 Thr Pro Glu Val Thr Cys Val
Val Val Asp Val Ser His Glu Asp Pro 260 265 270 Glu Val Lys Phe Asn
Trp Tyr Val Asp Gly Val Glu Val His Asn Ala 275 280 285
Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val 290
295 300 Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu
Tyr 305 310 315 320 Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro
Ile Glu Lys Thr 325 330 335 Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu
Pro Gln Val Tyr Thr Leu 340 345 350 Pro Pro Ser Arg Asp Glu Leu Thr
Lys Asn Gln Val Ser Leu Thr Cys 355 360 365 Leu Val Lys Gly Phe Tyr
Pro Ser Asp Ile Ala Val Glu Trp Glu Ser 370 375 380 Asn Gly Gln Pro
Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp 385 390 395 400 Ser
Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser 405 410
415 Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala
420 425 430 Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro
Gly Lys 435 440 445 <210> SEQ ID NO 84 <211> LENGTH:
216 <212> TYPE: PRT <213> ORGANISM: Artificial Sequence
<220> FEATURE: <221> NAME/KEY: source <223> OTHER
INFORMATION: /note="Description of Artificial Sequence: Synthetic
polypeptide" <400> SEQUENCE: 84 Ala Pro Glu Leu Leu Gly Gly
Pro Ser Val Phe Leu Phe Pro Pro Lys 1 5 10 15 Pro Lys Asp Thr Leu
Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val 20 25 30 Val Val Asp
Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr 35 40 45 Val
Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu 50 55
60 Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His
65 70 75 80 Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser
Asn Lys 85 90 95 Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys
Ala Lys Gly Gln 100 105 110 Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro
Pro Ser Arg Asp Glu Leu 115 120 125 Thr Lys Asn Gln Val Ser Leu Ser
Cys Ala Val Lys Gly Phe Tyr Pro 130 135 140 Ser Asp Ile Ala Val Glu
Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn 145 150 155 160 Tyr Lys Thr
Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu 165 170 175 Val
Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val 180 185
190 Phe Ser Cys Ser Val Leu His Glu Ala Leu His Ser His Tyr Thr Gln
195 200 205 Lys Ser Leu Ser Leu Ser Pro Gly 210 215
* * * * *