U.S. patent application number 17/173886 was filed with the patent office on 2021-09-09 for methods of treating conditions with antibodies that bind colony stimulating factor 1 receptor (csf1r).
This patent application is currently assigned to Five Prime Therapeutics, Inc.. The applicant listed for this patent is Five Prime Therapeutics, Inc.. Invention is credited to Haishan Lin, Emma Masteller, Brian Wong, Justin Wong.
Application Number | 20210277130 17/173886 |
Document ID | / |
Family ID | 1000005594424 |
Filed Date | 2021-09-09 |
United States Patent
Application |
20210277130 |
Kind Code |
A1 |
Wong; Brian ; et
al. |
September 9, 2021 |
Methods of Treating Conditions with Antibodies that Bind Colony
Stimulating Factor 1 Receptor (CSF1R)
Abstract
Methods of treating conditions with antibodies that bind colony
stimulating factor 1 receptor (CSF1R) are provided. Such methods
include, but are not limited to, methods of treating rheumatoid
arthritis and associated conditions, methods of treating systemic
lupus erythematosus and associated conditions, and methods of
treating multiple sclerosis.
Inventors: |
Wong; Brian; (Los Altos,
CA) ; Masteller; Emma; (Redwood City, CA) ;
Wong; Justin; (San Francisco, CA) ; Lin; Haishan;
(Moraga, CA) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
Five Prime Therapeutics, Inc. |
South San Francisco |
CA |
US |
|
|
Assignee: |
Five Prime Therapeutics,
Inc.
South San Francisco
CA
|
Family ID: |
1000005594424 |
Appl. No.: |
17/173886 |
Filed: |
February 11, 2021 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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15152161 |
May 11, 2016 |
10982001 |
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17173886 |
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13891455 |
May 10, 2013 |
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15152161 |
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61645901 |
May 11, 2012 |
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Current U.S.
Class: |
1/1 |
Current CPC
Class: |
A61K 39/3955 20130101;
C07K 2317/24 20130101; C07K 2317/55 20130101; C07K 2317/73
20130101; C07K 2317/51 20130101; C07K 2317/92 20130101; C07K
2317/515 20130101; C07K 16/2866 20130101; C07K 16/2869 20130101;
C07K 2317/622 20130101; C07K 2317/54 20130101; A61K 2039/505
20130101; C07K 2317/76 20130101; A61K 45/06 20130101; C07K 2317/565
20130101; C07K 2317/33 20130101 |
International
Class: |
C07K 16/28 20060101
C07K016/28; A61K 39/395 20060101 A61K039/395; A61K 45/06 20060101
A61K045/06 |
Claims
1.-38. (canceled)
39. A method of reducing the number of CD16+ monocytes in a subject
with a CD16+ disorder, comprising (a) determining the number of
CD16+ monocytes in a peripheral blood sample obtained from the
subject, and (b) administering to the subject an antibody that
binds human colony stimulating factor 1 receptor (CSF1R) such that
the number of CD16+ monocytes is reduced by at least 30%; wherein
the antibody (i) blocks binding of human colony stimulating factor
1 (CSF1) to human CSF1R and blocks binding of human IL-34 to human
CSF1R, and (ii) comprises a heavy chain comprising a heavy chain
(HC) CDR1 having the sequence of SEQ ID NO: 15, an HC CDR2 having
the sequence of SEQ ID NO: 16, and an HC CDR3 having the sequence
of SEQ ID NO: 17, and a light chain comprising a light chain (LC)
CDR1 having the sequence of SEQ ID NO: 18, a LC CDR2 having the
sequence of SEQ ID NO: 19, and a LC CDR3 having the sequence of SEQ
ID NO: 20.
40. The method of claim 39, further comprising determining the
number of CD16-monocytes in the sample, wherein the antibody is
administered such that the number of CD16- monocytes is not reduced
or is reduced by less than 20%.
41. The method of claim 39, further comprising (c) determining the
number of CD16+ monocytes in a further peripheral blood sample from
the subject after administration of the antibody.
42. The method of claim 41, further comprising determining the
number of CD16-monocytes in the samples of (a) and (c).
43. The method of claim 39, wherein the method further comprises
administering at least one additional therapeutic agent selected
from an anti-CD-20 antibody, an anti-CD19 antibody, an anti-GM-CSF
antibody, a CTLA-4 antagonist, and an anti-GM-CSF-R antibody.
44. The method of claim 39, comprising administering to the subject
the antibody that binds CSF1R such that the number of CD16+
monocytes is reduced by at least 50%.
45. The method of claim 39, wherein the antibody comprises a heavy
chain comprising the sequence of SEQ ID NO: 39 and a light chain
comprising the sequence of SEQ ID NO: 46.
46. The method of claim 39, wherein the antibody is a humanized
antibody.
47. The method of claim 39, wherein the antibody is a Fab, an Fv,
an scFv, a Fab', or a (Fab')2.
48. A method of reducing the number of CD16+ monocytes in a subject
with a CD16+ disorder, comprising (a) administering to the subject
an antibody that binds human colony stimulating factor 1 receptor
(CSF1R), (b) determining the number of CD16+ monocytes in a
peripheral blood sample obtained from the subject following
administration of the antibody, (c) determining whether
administration of the antibody reduces the number of CD16+
monocytes in the sample by at least 30% by comparing the number of
CD16+ monocytes in the sample of (b) with the number of CD16+
monocytes determined in a peripheral blood sample obtained from the
subject prior to administration, and optionally (d) adjusting the
dose or frequency of the administration of the antibody such that
the number of CD16+ monocytes remains reduced; wherein the antibody
(i) blocks binding of human colony stimulating factor 1 (CSF1) to
human CSF1R and blocks binding of human IL-34 to human CSF1R and
(ii) comprises a heavy chain comprising a heavy chain (HC) CDR1
having the sequence of SEQ ID NO: 15, an HC CDR2 having the
sequence of SEQ ID NO: 16, and an HC CDR3 having the sequence of
SEQ ID NO: 17, and a light chain comprising a light chain (LC) CDR1
having the sequence of SEQ ID NO: 18, a LC CDR2 having the sequence
of SEQ ID NO: 19, and a LC CDR3 having the sequence of SEQ ID NO:
20.
49. The method of claim 48, comprising adjusting the dose or
frequency of the administration of the antibody such that the
number of CD16+ monocytes remains reduced by at least 30%.
50. The method of claim 48, further comprising determining the
number of CD16-monocytes in the sample after administration,
wherein the antibody is administered such that the number of CD16-
monocytes is not reduced or is reduced by less than 20%.
51. The method of claim 48, wherein the method further comprises
administering at least one additional therapeutic agent selected
from an anti-CD-20 antibody, an anti-CD19 antibody, an anti-GM-CSF
antibody, a CTLA-4 antagonist, and an anti-GM-CSF-R antibody.
52. The method of claim 48, comprising determining whether
administration of the antibody reduces the number of CD16+
monocytes in the sample by at least 50%, and optionally adjusting
the dose or frequency of the administration of the antibody such
that the number of CD16+ monocytes remains reduced by at least
50%.
53. The method of claim 48, wherein the antibody comprises a heavy
chain comprising the sequence of SEQ ID NO: 39 and a light chain
comprising the sequence of SEQ ID NO: 46.
54. The method of claim 48, wherein the antibody is a humanized
antibody.
55. The method of claim 48, wherein the antibody is a Fab, an Fv,
an scFv, a Fab', or a (Fab')2.
56. A method of treating neoplasia in a subject, comprising (a)
determining the number of CD16+ monocytes in a peripheral blood
sample obtained from the subject, and (b) administering to the
subject an antibody that binds human colony stimulating factor 1
receptor (CSF1R) such that the number of CD16+ monocytes is reduced
by at least 30%; wherein the antibody (i) blocks binding of human
colony stimulating factor 1 (CSF1) to human CSF1R and blocks
binding of human IL-34 to human CSF1R, and (ii) comprises a heavy
chain comprising a heavy chain (HC) CDR1 having the sequence of SEQ
ID NO: 15, an HC CDR2 having the sequence of SEQ ID NO: 16, and an
HC CDR3 having the sequence of SEQ ID NO: 17, and a light chain
comprising a light chain (LC) CDR1 having the sequence of SEQ ID
NO: 18, a LC CDR2 having the sequence of SEQ ID NO: 19, and a LC
CDR3 having the sequence of SEQ ID NO: 20.
57. The method of claim 56, wherein the antibody comprises a heavy
chain comprising the sequence of SEQ ID NO: 39 and a light chain
comprising the sequence of SEQ ID NO: 46.
58. The method of claim 56, further comprising (c) determining the
number of CD16+ monocytes in a further peripheral blood sample from
the subject after administration of the antibody.
Description
[0001] This application is a divisional of U.S. application Ser.
No. 15/152,161, filed May 11, 2016, which is a divisional of U.S.
application Ser. No. 13/891,455, filed May 10, 2013, now abandoned,
which claims the benefit of U.S. Provisional Application No.
61/645,901, filed May 11, 2012, which is incorporated by reference
herein in its entirety for any purpose.
TECHNICAL FIELD
[0002] Methods of treating conditions with antibodies that bind
colony stimulating factor 1 receptor (CSF1R) are provided. Such
methods include, but are not limited to, methods of treating
rheumatoid arthritis, multiple sclerosis, and systemic lupus
erythematosus.
BACKGROUND
[0003] Colony stimulating factor 1 receptor (referred to herein as
CSF1R; also referred to in the art as FMS, FIM2, C-FMS, M-CSF
receptor, and CD115) is a single-pass transmembrane receptor with
an N-terminal extracellular domain (ECD) and a C-terminal
intracellular domain with tyrosine kinase activity. Ligand binding
of CSF1 or the interleukin 34 ligand (referred to herein as IL-34;
Lin et al., Science 320: 807-11 (2008)) to CSF1R leads to receptor
dimerization, upregulation of CSF1R protein tyrosine kinase
activity, phosphorylation of CSF1R tyrosine residues, and
downstream signaling events. Both CSF1 and IL-34 stimulate monocyte
survival, proliferation, and differentiation into macrophages, as
well as other monocytic cell lineages such as osteoclasts,
dendritic cells, and microglia.
[0004] Many tumor cells have been found to secrete CSF1, which
activates monocyte/macrophage cells through CSF1R. The level of
CSF1 in tumors has been shown to correlate with the level of
tumor-associated macrophages (TAMs) in the tumor. Higher levels of
TAMs have been found to correlate with poorer patient prognoses. In
addition, CSF1 has been found to promote tumor growth and
progression to metastasis in, for example, human breast cancer
xenografts in mice. See, e.g., Paulus et al., Cancer Res. 66:
4349-56 (2006). Further, CSF1R plays a role in osteolytic bone
destruction in bone metastasis. See, e.g., Ohno et al., Mol. Cancer
Ther. 5: 2634-43 (2006).
[0005] CSF1 and its receptor have also been found to be involved in
various inflammatory and autoimmune diseases. See, e.g., Hamilton,
Nat. Rev. 8: 533-44 (2008). For example, synovial endothelial cells
from joints afflicted with rheumatoid arthritis have been found to
produce CSF1, suggesting a role for CSF1 and its receptor in the
disease. Blocking CSF1R activity with an antibody results in
positive clinical effects in mouse models of arthritis, including a
reduction in the destruction of bone and cartilage and a reduction
in macrophage numbers. See, e.g., Kitaura et al., J. Clin. Invest.
115: 3418-3427 (2005).
[0006] Mature differentiated myeloid lineage cells such as
macrophages, microglial cells, and osteoclasts contribute to
pathology of various diseases such as rheumatoid arthritis,
multiple sclerosis and diseases of bone loss. Differentiated
myeloid lineage cells are derived from peripheral blood monocyte
intermediates. CSF1R stimulation contributes to development of
monocytes from bone marrow precursors, to monocyte proliferation
and survival, and to differentiation of peripheral blood monocytes
into differentiated myeloid lineage cells such as macrophages,
microglial cells, and osteoclasts. CSF1R stimulation thus
contributes to proliferation, survival, activation, and maturation
of differentiated myeloid lineage cells, and in the pathologic
setting, CSF1R stimulation contributes to the ability of
differentiated myeloid lineage cells to mediate disease
pathology.
SUMMARY
[0007] In some embodiments, a method of treating a condition
associated with rheumatoid arthritis is provided. In some
embodiments, the method comprises administering an antibody that
binds colony stimulating factor 1 receptor (CSF1R) to a subject
with rheumatoid arthritis, wherein the antibody blocks binding of
colony stimulating factor 1 (CSF1) to CSF1R and blocks binding of
IL-34 to CSF1R. In some embodiments, treating a condition
associated with rheumatoid arthritis comprises at least one effect
selected from reducing inflammation, reducing pannus formation,
reducing cartilage damage, reducing bone resorption, reducing the
number of macrophages in at least one joint affected by rheumatoid
arthritis, reducing autoantibody levels, and reducing bone loss. In
some embodiments, treating a condition associated with rheumatoid
arthritis comprises reducing inflammation. In some embodiments, a
method of reducing inflammation associated with rheumatoid
arthritis comprises determining an erythrocyte sedimentation rate,
wherein a reduced sedimentation rate indicates reduced
inflammation.
[0008] In some embodiments, a method of treating a condition
associated with rheumatoid arthritis comprises at least one effect
selected from reducing pannus formation, reducing bone resorption,
and reducing bone loss. In some embodiments, treating a condition
associated with rheumatoid arthritis comprises reducing bone
resorption. In some such embodiments, the level of at least one
marker of bone resorption is reduced. In some embodiments, the bone
resorption marker is selected from tartrate resistant acid
phosphatase 5b (TRAP5b), urinary total pyridinoline, Urinary total
deoxypyridinoline, urinary free pyridinoline, serum collagen type I
cross-linked N-telopeptide, urinary collagen type I cross-linked
N-telopeptide, and serum carboxyterminal telopeptide of type I
collagen.
[0009] In some embodiments, treating a condition associated with
rheumatoid arthritis comprises at least one effect selected from
reducing pannus formation, and reducing bone loss. In some
embodiments, the at least one effect is measured using an imaging
technique. In some such embodiments, the imaging technique
comprises a method selected from x-ray imaging, magnetic resonance
imaging, computed tomography (CT) scan, arthroscopy, scintigraphy,
ultrasonography, bone densitometry, single photon absorptiometry
(SPA), dual photon absorptiometry (DPA), single energy x-ray
absorptiometry (SXA), dual energy x-ray absorptiometry (DXA),
scintigraphy, ultrasonography, duplex ultrasonography, and power
doppler imaging.
[0010] In some embodiments of a method of treating a condition
associated with rheumatoid arthritis, the number of CD16+ monocytes
is reduced by at least 30%. In some embodiments of the method, the
number of CD16- monocytes is not reduced or is reduced by less than
20%. In some embodiments, the CD16+ monocytes are CD16+ peripheral
blood monocytes.
[0011] In some embodiments, a method of treating rheumatoid
arthritis is provided, comprising administering an antibody that
binds CSF1R to a subject with rheumatoid arthritis, wherein the
antibody blocks binding of CSF1 to CSF1R and blocks binding of
IL-34 to CSF1R, and wherein the antibody reduces the number of
CD16+ monocytes in the subject by at least 30%, and wherein CD16-
monocytes are not reduced or are reduced by less than 20%. In some
embodiments, the CD16+ monocytes are CD16+ peripheral blood
monocytes.
[0012] In some embodiments of a method of treating rheumatoid
arthritis, the method further comprises administering at least one
additional therapeutic agent selected from methotrexate, an
anti-TNF agent, a glucocorticoid, cyclosporine, leflunomide,
azathioprine, a JAK inhibitor, a SYK inhibitor, an anti-IL-6
antibody, an anti-IL-6R antibody, an anti-CD-20 antibody, an
anti-CD19 antibody, an anti-GM-CSF antibody, an IL-1 receptor
antagonist, a CTLA-4 antagonist, and an anti-GM-CSF-R antibody.
[0013] In some embodiments, methods of treating skin lesions
associated with lupus are provided. In some embodiments, the method
comprises administering an antibody that binds CSF1R to a subject
with lupus, wherein the antibody blocks binding of CSF1 to CSF1R
and blocks binding of IL-34 to CSF1R. In some embodiments, treating
skin lesions associated with lupus comprises at least one effect
selected from reducing the number of skin lesions, reducing the
rate of formation of skin lesions, and reducing the severity of
skin lesions.
[0014] In some embodiments, methods of treating lupus nephritis are
provided. In some embodiments, the method comprises administering
an antibody that binds CSF1R to a subject with lupus nephritis,
wherein the antibody blocks binding of CSF1 to CSF1R and blocks
binding of IL-34 to CSF1R. In some embodiments, kidney function is
improved in the subject. In some embodiments, proteinuria is
reduced in the subject. In some embodiments, glomerular filtration
rate is improved in the subject.
[0015] In some embodiments, methods of treating lupus are provided.
In some embodiments, a method comprises administering an antibody
that binds CSF1R to a subject with lupus, wherein the antibody
blocks binding of CSF1 to CSF1R and blocks binding of IL-34 to
CSF1R. In some embodiments, the antibody reduces the number of
CD16+ monocytes in the subject by at least 30%, and CD16- monocytes
are not reduced or are reduced by less than 20%. In some
embodiments, CD16+ monocytes are reduced by at least 50%. In some
embodiments, the CD16+ monocytes are CD16+ peripheral blood
monocytes.
[0016] In some embodiments, the method of treating lupus, lupus
nephritis, or skin lesions associated with lupus further comprises
administering at least one additional therapeutic agent selected
from hydroxychloroquine (Plaquenil), a corticosteroids,
cyclophosphamide (Cytoxan), azathioprine (Imuran, Azasan),
mycophenolate (Cellcept), leflunomide (Arava) and methotrexate
(Trexall), and belimumab (Benlysta).
[0017] In some embodiments, methods of treating inflammatory
conditions are provided. In some such embodiments, a method
comprises administering an antibody that binds CSF1R to a subject
with an inflammatory condition, wherein the antibody blocks binding
of CSF1 to CSF1R and blocks binding of IL-34 to CSF1R. In some
embodiments, the antibody reduces the number of CD16+ monocytes by
at least 30%, and CD16-monocytes are not reduced or are reduced by
less than 20%. In some embodiments, CD16+ monocytes are reduced by
at least 50%. In some embodiments, the CD16+ monocytes are CD16+
peripheral blood monocytes.
[0018] In some embodiments, methods of treating CD16+ disorders are
provided. In some embodiments, a method comprises administering an
antibody that binds CSF1R to a subject with a CD16+ disorder,
wherein the antibody blocks binding of CSF1 to CSF1R and blocks
binding of IL-34 to CSF1R. In some embodiments, the antibody
reduces the number of CD16+ monocytes by at least 30%, and CD16-
monocytes are not reduced or are reduced by less than 20%. In some
embodiments, CD16+ monocytes are reduced by at least 50%. In some
embodiments, the CD16+ monocytes are CD16+ peripheral blood
monocytes. In some embodiments, the CD16+ disorder is selected from
rheumatoid arthritis, juvenile idiopathic arthritis, psoriasis,
psoriatic arthritis, ankylosing spondylitis, Crohn's disease, and
ulcerative colitis, lupus erythematosus, inflammatory bowel
disease, Sjogren's syndrome, coronary artery disease, asthma, and
sarcoidosis.
[0019] In some embodiments, a method of treating discussed herein
comprises treating a condition that does not respond to
methotrexate.
[0020] In some embodiments, methods of reducing the number of CD16+
monocytes are provided. In some embodiments, a method comprises
administering an antibody that binds CSF1R to a subject, wherein
the antibody blocks binding of CSF1 to CSF1R and blocks binding of
IL-34 to CSF1R. In some embodiments, the antibody reduces the
number of CD16+ monocytes by at least 30%, and CD16- monocytes are
not reduced or are reduced by less than 20%. In some embodiments,
CD16+ monocytes are reduced by at least 50%. In some embodiments,
the CD16+ monocytes are CD16+ peripheral blood monocytes.
[0021] In some embodiments, methods of slowing the progression of a
kidney condition associated with lupus are provided. In some
embodiments, the method comprises administering an antibody that
binds CSF1R to a subject, wherein the antibody blocks binding of
CSF1 to CSF1R and blocks binding of IL-34 to CSF1R. In some
embodiments, proteinuria does not increase in the subject or does
not increase in the subject at the same rate as in subjects not
administered the antibody. In some embodiments, glomerular
filtration rate does not decrease in the subject or does not
decrease in the subject at the same rate as in a subjects not
administered the antibody.
[0022] In some embodiments, methods of slowing the progression of
pannus formation in a subject with rheumatoid arthritis are
provided. In some such embodiments, a method comprises
administering an antibody that binds CSF1R to a subject with
rheumatoid arthritis, wherein the antibody blocks binding of CSF1
to CSF1R and blocks binding of IL-34 to CSF1R.
[0023] In some embodiments, methods of slowing the progression of
bone loss in a subject with rheumatoid arthritis are provided. In
some embodiments, a method comprises administering an antibody that
binds colony stimulating factor 1 receptor (CSF1R) to a subject
with rheumatoid arthritis, wherein the antibody blocks binding of
colony stimulating factor 1 (CSF1) to CSF1R and blocks binding of
IL-34 to CSF1R.
[0024] In some embodiments of the methods described herein, the
antibody heavy chain and/or the antibody light chain have the
following structure.
[0025] In some embodiments, the heavy chain comprises a sequence
that is at least 90%, at least 95%, at least 97%, at least 99%, or
100% identical to a sequence selected from SEQ ID NOs: 9, 11, 13,
and 39 to 45. In some embodiments, the light chain comprises a
sequence that is at least 90%, at least 95%, at least 97%, at least
99%, or 100% identical to a sequence selected from SEQ ID NOs: 10,
12, 14, and 46 to 52. In some embodiments, the heavy chain
comprises a sequence that is at least 90%, at least 95%, at least
97%, at least 99%, or 100% identical to a sequence selected from
SEQ ID NOs: 9, 11, 13, and 39 to 45, and the light chain comprises
a sequence that is at least 90%, at least 95%, at least 97%, at
least 99%, or 100% identical to a sequence selected from SEQ ID
NOs: 10, 12, 14, and 46 to 52.
[0026] In some embodiments, the HC CDR1, HC CDR2, and HC CDR3
comprise a set of sequences selected from: (a) SEQ ID NOs: 15, 16,
and 17; (b) SEQ ID NOs: 21, 22, and 23; and (c) SEQ ID NOs: 27, 28,
and 29. In some embodiments, the LC CDR1, LC CDR2, and LC CDR3
comprise a set of sequences selected from: (a) SEQ ID NOs: 18, 19,
and 20; (b) SEQ ID NOs: 24, 25, and 26; and (c) SEQ ID NOs: 30, 31,
and 32.
[0027] In some embodiments, the heavy chain comprises an HC CDR1,
HC CDR2, and HC CDR3, wherein the HC CDR1, HC CDR2, and HC CDR3
comprise a set of sequences selected from: (a) SEQ ID NOs: 15, 16,
and 17; (b) SEQ ID NOs: 21, 22, and 23; and (c) SEQ ID NOs: 27, 28,
and 29; and the light chain comprises an LC CDR1, LC CDR2, and LC
CDR3, wherein the LC CDR1, LC CDR2, and LC CDR3 comprise a set of
sequences selected from: (a) SEQ ID NOs: 18, 19, and 20; (b) SEQ ID
NOs: 24, 25, and 26; and (c) SEQ ID NOs: 30, 31, and 32.
[0028] In some embodiments, an isolated antibody is provided,
wherein the antibody comprises a heavy chain and a light chain,
wherein the antibody comprises: (a) a heavy chain comprising a
sequence that is at least 95%, at least 97%, at least 99%, or 100%
identical to SEQ ID NO: 9 and a light chain comprising a sequence
that is at least 95%, at least 97%, at least 99%, or 100% identical
to SEQ ID NO: 10; (b) a heavy chain comprising a sequence that is
at least 95%, at least 97%, at least 99%, or 100% identical to SEQ
ID NO: 11 and a light chain comprising a sequence that is at least
95%, at least 97%, at least 99%, or 100% identical to SEQ ID NO:
12; (c) a heavy chain comprising a sequence that is at least 95%,
at least 97%, at least 99%, or 100% identical to SEQ ID NO: 13 and
a light chain comprising a sequence that is at least 95%, at least
97%, at least 99%, or 100% identical to SEQ ID NO: 14; (d) a heavy
chain comprising a sequence that is at least 95%, at least 97%, at
least 99%, or 100% identical to SEQ ID NO: 39 and a light chain
comprising a sequence that is at least 95%, at least 97%, at least
99%, or 100% identical to SEQ ID NO: 46; (e) a heavy chain
comprising a sequence that is at least 95%, at least 97%, at least
99%, or 100% identical to SEQ ID NO: 40 and a light chain
comprising a sequence that is at least 95%, at least 97%, at least
99%, or 100% identical to SEQ ID NO: 46; (f) a heavy chain
comprising a sequence that is at least 95%, at least 97%, at least
99%, or 100% identical to SEQ ID NO: 41 and a light chain
comprising a sequence that is at least 95%, at least 97%, at least
99%, or 100% identical to SEQ ID NO: 46; (g) a heavy chain
comprising a sequence that is at least 95%, at least 97%, at least
99%, or 100% identical to SEQ ID NO: 39 and a light chain
comprising a sequence that is at least 95%, at least 97%, at least
99%, or 100% identical to SEQ ID NO: 47; (h) a heavy chain
comprising a sequence that is at least 95%, at least 97%, at least
99%, or 100% identical to SEQ ID NO: 40 and a light chain
comprising a sequence that is at least 95%, at least 97%, at least
99%, or 100% identical to SEQ ID NO: 47; (i) a heavy chain
comprising a sequence that is at least 95%, at least 97%, at least
99%, or 100% identical to SEQ ID NO: 41 and a light chain
comprising a sequence that is at least 95%, at least 97%, at least
99%, or 100% identical to SEQ ID NO: 47; and (j) a heavy chain
comprising a sequence that is at least 95%, at least 97%, at least
99%, or 100% identical to SEQ ID NO: 42 and a light chain
comprising a sequence that is at least 95%, at least 97%, at least
99%, or 100% identical to SEQ ID NO: 48; (k) a heavy chain
comprising a sequence that is at least 95%, at least 97%, at least
99%, or 100% identical to SEQ ID NO: 42 and a light chain
comprising a sequence that is at least 95%, at least 97%, at least
99%, or 100% identical to SEQ ID NO: 49; (1) a heavy chain
comprising a sequence that is at least 95%, at least 97%, at least
99%, or 100% identical to SEQ ID NO: 42 and a light chain
comprising a sequence that is at least 95%, at least 97%, at least
99%, or 100% identical to SEQ ID NO: 50; (m) a heavy chain
comprising a sequence that is at least 95%, at least 97%, at least
99%, or 100% identical to SEQ ID NO: 43 and a light chain
comprising a sequence that is at least 95%, at least 97%, at least
99%, or 100% identical to SEQ ID NO: 48; (n) a heavy chain
comprising a sequence that is at least 95%, at least 97%, at least
99%, or 100% identical to SEQ ID NO: 43 and a light chain
comprising a sequence that is at least 95%, at least 97%, at least
99%, or 100% identical to SEQ ID NO: 49; (o) a heavy chain
comprising a sequence that is at least 95%, at least 97%, at least
99%, or 100% identical to SEQ ID NO: 43 and a light chain
comprising a sequence that is at least 95%, at least 97%, at least
99%, or 100% identical to SEQ ID NO: 50; (p) a heavy chain
comprising a sequence that is at least 95%, at least 97%, at least
99%, or 100% identical to SEQ ID NO: 44 and a light chain
comprising a sequence that is at least 95%, at least 97%, at least
99%, or 100% identical to SEQ ID NO: 51; (q) a heavy chain
comprising a sequence that is at least 95%, at least 97%, at least
99%, or 100% identical to SEQ ID NO: 44 and a light chain
comprising a sequence that is at least 95%, at least 97%, at least
99%, or 100% identical to SEQ ID NO: 52; (r) a heavy chain
comprising a sequence that is at least 95%, at least 97%, at least
99%, or 100% identical to SEQ ID NO: 45 and a light chain
comprising a sequence that is at least 95%, at least 97%, at least
99%, or 100% identical to SEQ ID NO: 51; or (s) a heavy chain
comprising a sequence that is at least 95%, at least 97%, at least
99%, or 100% identical to SEQ ID NO: 45 and a light chain
comprising a sequence that is at least 95%, at least 97%, at least
99%, or 100% identical to SEQ ID NO: 52.
[0029] In some embodiments, an antibody is provided, wherein the
antibody comprises a heavy chain and a light chain, wherein the
antibody comprises: (a) a heavy chain comprising a heavy chain (HC)
CDR1 having the sequence of SEQ ID NO: 15, an HC CDR2 having the
sequence of SEQ ID NO: 16, and an HC CDR3 having the sequence of
SEQ ID NO: 17, and a light chain comprising a light chain (LC) CDR1
having the sequence of SEQ ID NO: 18, a LC CDR2 having the sequence
of SEQ ID NO: 19, and a LC CDR3 having the sequence of SEQ ID NO:
20; (b) a heavy chain comprising a heavy chain (HC) CDR1 having the
sequence of SEQ ID NO: 21, an HC CDR2 having the sequence of SEQ ID
NO: 22, and an HC CDR3 having the sequence of SEQ ID NO: 23, and a
light chain comprising a light chain (LC) CDR1 having the sequence
of SEQ ID NO: 24, a LC CDR2 having the sequence of SEQ ID NO: 25,
and a LC CDR3 having the sequence of SEQ ID NO: 26; or (c) a heavy
chain comprising a heavy chain (HC) CDR1 having the sequence of SEQ
ID NO: 27, an HC CDR2 having the sequence of SEQ ID NO: 28, and an
HC CDR3 having the sequence of SEQ ID NO: 29, and a light chain
comprising a light chain (LC) CDR1 having the sequence of SEQ ID
NO: 30, a LC CDR2 having the sequence of SEQ ID NO: 31, and a LC
CDR3 having the sequence of SEQ ID NO: 32.
[0030] In some embodiments, an antibody comprises a heavy chain and
a light chain, wherein the antibody comprises: (a) a heavy chain
comprising a sequence of SEQ ID NO: 53 and a light chain comprising
a sequence of SEQ ID NO: 60; (b) a heavy chain comprising a
sequence of SEQ ID NO: 53 and a light chain comprising a sequence
of SEQ ID NO: 61; or (c) a heavy chain comprising a sequence of SEQ
ID NO: 58 and a light chain comprising a sequence of SEQ ID NO: 65.
In some embodiments, an antibody comprises a heavy chain and a
light chain, wherein the antibody comprises: (a) a heavy chain
consisting of the sequence of SEQ ID NO: 53 and a light chain
consisting of the sequence of SEQ ID NO: 60; (b) a heavy chain
consisting of the sequence of SEQ ID NO: 53 and a light chain
consisting of the sequence of SEQ ID NO: 61; or (c) a heavy chain
consisting of the sequence of SEQ ID NO: 58 and a light chain
consisting of the sequence of SEQ ID NO: 65.
[0031] In some embodiments, an antibody is a humanized antibody. In
some embodiments, an antibody is selected from a Fab, an Fv, an
scFv, a Fab', and a (Fab').sub.2. In some embodiments, an antibody
is a chimeric antibody. In some embodiments, an antibody is
selected from an IgA, an IgG, and an IgD. In some embodiments, an
antibody is an IgG. In some embodiments, an antibody is an IgG4. In
some embodiments, an antibody is an IgG4 comprising an S241P
mutation in at least one IgG4 heavy chain constant region.
[0032] In some embodiments, an antibody binds to human CSF1R and/or
binds to cynomolgus CSF1R. In some embodiments, an antibody blocks
ligand binding to CSF1R. In some embodiments, an antibody blocks
binding of CSF1 and/or IL-34 to CSF1R. In some embodiments, an
antibody blocks binding of both CSF1 and IL-34 to CSF1R. In some
embodiments, an antibody inhibits ligand-induced CSF1R
phosphorylation. In some embodiments, an antibody inhibits CSF1-
and/or IL-34-induced CSF1R phosphorylation. In some embodiments, an
antibody binds to human CSF1R with an affinity (K.sub.D) of less
than 1 nM. In some embodiments, antibody inhibits monocyte
proliferation and/or survival responses in the presence of CSF1 or
IL-34.
[0033] In some embodiments, a pharmaceutical composition comprising
an antibody that binds CSF1R is provided.
[0034] In some embodiments, compositions comprising antibodies that
bind CSF1R are provided for use in methods of treatment of human or
animals. In some embodiments, antibodies that bind CSF1R and
compositions comprising antibodies that bind CSF1R are provided for
use in a method of treating conditions associated with rheumatoid
arthritis in a human or animal. In some embodiments, antibodies
that bind CSF1R and compositions comprising antibodies that bind
CSF1R are provided for use in a method of treating multiple
sclerosis in a human or animal. In some embodiments, antibodies
that bind CSF1R and compositions comprising antibodies that bind
CSF1R are provided for use in a method of treating conditions
associated with systemic lupus erythematosus are provided.
BRIEF DESCRIPTION OF THE FIGURES
[0035] FIG. 1A-C shows an alignment of the humanized heavy chain
variable regions for each of humanized antibodies huAb1 to huAb16,
as discussed in Example 1. Boxed residues are amino acids in the
human acceptor sequence that were changed back to the corresponding
mouse residue.
[0036] FIG. 2A-C shows an alignment of the humanized light chain
variable regions for each of humanized antibodies huAb1 to huAb16,
as discussed in Example 1 Boxed amino acids are residues in the
human acceptor sequence that were changed back to the corresponding
mouse residue.
[0037] FIG. 3 shows reduction in CD16+ monocytes in cynomolgus
monkeys administered huAb1, as described in Example 2.
[0038] FIG. 4 shows dose-dependent binding of surrogate antibody
cAb1 to mouse CSF1R, as described in Example 3.
[0039] FIG. 5 shows dose-dependent inhibition of CSF1- and
IL-34-induced proliferation in mNFS60 cells, as described in
Example 4.
[0040] FIG. 6 shows suppression of clinical disease scores in a
mouse model of rheumatoid arthritis following administration of
cAb1, as described in Example 5.
[0041] FIG. 7 shows suppression of bone loss, indicated by plasma
TRAP5b levels, in a mouse model of rheumatoid arthritis following
administration of cAb1, as described in Example 6.
[0042] FIG. 8 shows suppression of inflammation, pannus formation,
cartilage damage, and bone damage in a mouse model of rheumatoid
arthritis following administration of cAb1, as described in Example
7.
[0043] FIG. 9 shows suppression of macrophage numbers in paw joints
and knee joints in a mouse model of rheumatoid arthritis following
administration of cAb1, as described in Example 8.
[0044] FIG. 10 shows suppression of autoantibody formation in a
mouse model of rheumatoid arthritis following administration of
cAb1, as described in Example 9.
[0045] FIG. 11 shows suppression of bone loss in a mouse model of
established rheumatoid arthritis following administration of cAb1,
as described in Example 10.
[0046] FIG. 12 shows suppression of pannus formation and bone
destruction in a mouse model of established rheumatoid arthritis
following administration of cAb1, as described in Example 11.
[0047] FIG. 13 shows suppression of glomerulonephritis,
interstitial nephritis, and perivascular infiltrates in a mouse
model of systemic lupus erythematosus following administration of
cAb1, as described in Example 12.
[0048] FIG. 14 shows suppression of skin lesions in a mouse model
of systemic lupus erythematosus following administration of cAb1,
as described in Example 13.
[0049] FIG. 15 shows suppression of clinical disease scores in a
mouse model of multiple sclerosis following administration of cAb1,
as described in Example 14.
DETAILED DESCRIPTION
[0050] Methods of treating conditions comprising administering
antibodies that bind CSF1R and block CSF1 and IL-34 ligand binding
are provided. As discussed herein, antibodies that bind CSF1R and
block CSF1 and IL-34 ligand binding are effective for treating
rheumatoid arthritis, systemic lupus erythematosus, and multiple
sclerosis. The present inventors found that administering such
antibodies to cynomolgus monkeys reduced the number of CD16+
peripheral blood monocytes in cynomolgus monkeys, but does not
affect CD16- peripheral blood monocyte numbers CD16+ peripheral
blood monocytes are highly inflammatory monocytes. See, e.g.,
Ziegler-Heitbrock, J. Leukocyte Biol., 2007, 81: 584-592. Further,
administering such antibodies in a mouse model of rheumatoid
arthritis suppressed clinical disease scores, including suppression
of erythema and swelling; suppressed bone loss as measured by a
reduction in tartrate resistant acid phosphatase 5b (TRAP5b)
levels; and suppressed inflammation, cartilage destruction, pannus
formation, and bone destruction. Notably, when the antibodies were
administered after the appearance of clinical manifestations of
rheumatoid arthritis, bone loss (as measured by a reduction in
TRAP5b levels), pannus formation, and bone destruction were
reduced. Administering such antibodies in a mouse model of
rheumatoid arthritis also reduced joint macrophage numbers and
autoantibody formation.
[0051] Administering antibodies that bind CSF1R and block CSF1 and
IL-34 ligand binding in a systemic lupus erythematosus mouse model
suppressed glomerulonephritis, interstitial nephritis, and
perivascular infiltrates. Further, administering such antibodies in
a systemic lupus erythematosus mouse model suppressed skin lesions.
Finally, administering the antibodies in a mouse model of multiple
sclerosis suppressed clinical disease scores of MS.
[0052] The section headings used herein are for organizational
purposes only and are not to be construed as limiting the subject
matter described. All references cited herein, including patent
applications and publications, are incorporated by reference in
their entirety.
Definitions
[0053] Unless otherwise defined, scientific and technical terms
used in connection with the present invention shall have the
meanings that are commonly understood by those of ordinary skill in
the art. Further, unless otherwise required by context, singular
terms shall include pluralities and plural terms shall include the
singular.
[0054] Exemplary techniques used in connection with recombinant
DNA, oligonucleotide synthesis, tissue culture and transformation
(e.g., electroporation, lipofection), enzymatic reactions, and
purification techniques are known in the art. Many such techniques
and procedures are described, e.g., in Sambrook et al. Molecular
Cloning: A Laboratory Manual (2nd ed., Cold Spring Harbor
Laboratory Press, Cold Spring Harbor, N.Y. (1989)), among other
places. In addition, exemplary techniques for chemical syntheses,
chemical analyses, pharmaceutical preparation, formulation, and
delivery, and treatment of patients are also known in the art.
[0055] In this application, the use of "or" means "and/or" unless
stated otherwise. In the context of a multiple dependent claim, the
use of "or" refers back to more than one preceding independent or
dependent claim in the alternative only. Also, terms such as
"element" or "component" encompass both elements and components
comprising one unit and elements and components that comprise more
than one subunit unless specifically stated otherwise.
[0056] As utilized in accordance with the present disclosure, the
following terms, unless otherwise indicated, shall be understood to
have the following meanings:
[0057] The terms "nucleic acid molecule" and "polynucleotide" may
be used interchangeably, and refer to a polymer of nucleotides.
Such polymers of nucleotides may contain natural and/or non-natural
nucleotides, and include, but are not limited to, DNA, RNA, and
PNA. "Nucleic acid sequence" refers to the linear sequence of
nucleotides that comprise the nucleic acid molecule or
polynucleotide.
[0058] The terms "polypeptide" and "protein" are used
interchangeably to refer to a polymer of amino acid residues, and
are not limited to a minimum length. Such polymers of amino acid
residues may contain natural or non-natural amino acid residues,
and include, but are not limited to, peptides, oligopeptides,
dimers, trimers, and multimers of amino acid residues. Both
full-length proteins and fragments thereof are encompassed by the
definition. The terms also include post-expression modifications of
the polypeptide, for example, glycosylation, sialylation,
acetylation, phosphorylation, and the like. Furthermore, for
purposes of the present invention, a "polypeptide" refers to a
protein which includes modifications, such as deletions, additions,
and substitutions (generally conservative in nature), to the native
sequence, as long as the protein maintains the desired activity.
These modifications may be deliberate, as through site-directed
mutagenesis, or may be accidental, such as through mutations of
hosts which produce the proteins or errors due to PCR
amplification.
[0059] The term "CSF1R" refers herein to the full-length CSF1R,
which includes the N-terminal ECD, the transmembrane domain, and
the intracellular tyrosine kinase domain, with or without an
N-terminal leader sequence. In some embodiments, the CSF1R is a
human CSF1R having the amino acid sequence of SEQ ID NO: 1 or SEQ
ID NO: 2.
[0060] The term "CSF1R extracellular domain" ("CSF1R ECD") as used
herein refers to a CSF1R polypeptide that lacks the intracellular
and transmembrane domains. CSF1R ECDs include the full-length CSF1R
ECD and CSF1R ECD fragments that are capable of binding CSF1R
and/or IL-34. The human full-length CSF1R ECD is defined herein as
comprising either amino acids 1 to 512 (i.e., including the leader
sequence) or amino acids 20 to 512 (i.e., lacking the leader
sequence) of SEQ ID NO: 2. In some embodiments, a human CSF1R ECD
fragment comprises amino acids 20 to 506 of SEQ ID NO: 2 (see SEQ
ID NO:5). In some embodiments, a human CSF1R fragment ends at amino
acid 507, 508, 509, 510, or 511. In some embodiments, a cyno CSF1R
ECD comprises the sequence of SEQ ID NO: 7 (with leader sequence)
or amino acids 20 to 506 of SEQ ID NO: 7 (without leader
sequence).
[0061] With reference to anti-CSF1R antibodies the terms "active"
or "activity" or "function", and grammatical variants thereof, are
used to refer to the ability to inhibit (blocking or antagonist
antibodies) or mimic (agonist antibodies) at least one of the
foregoing activities. Antibodies and antibody fragments referred to
as "functional" are characterized by having such properties.
[0062] An "immunological" activity refers only to the ability to
induce the production of an antibody against an antigenic epitope
possessed by a native or naturally-occurring CSF1R polypeptide.
[0063] The term "antibody" as used herein refers to a molecule
comprising at least complementarity-determining region (CDR) 1,
CDR2, and CDR3 of a heavy chain and at least CDR1, CDR2, and CDR3
of a light chain, wherein the molecule is capable of binding to
antigen. The term antibody includes, but is not limited to,
fragments that are capable of binding antigen, such as Fv,
single-chain Fv (scFv), Fab, Fab', and (Fab').sub.2. The term
antibody also includes, but is not limited to, chimeric antibodies,
humanized antibodies, and antibodies of various species such as
mouse, human, cynomolgus monkey, etc.
[0064] In some embodiments, an antibody comprises a heavy chain
variable region and a light chain variable region. In some
embodiments, an antibody comprises at least one heavy chain
comprising a heavy chain variable region and at least a portion of
a heavy chain constant region, and at least one light chain
comprising a light chain variable region and at least a portion of
a light chain constant region. In some embodiments, an antibody
comprises two heavy chains, wherein each heavy chain comprises a
heavy chain variable region and at least a portion of a heavy chain
constant region, and two light chains, wherein each light chain
comprises a light chain variable region and at least a portion of a
light chain constant region. As used herein, a single-chain Fv
(scFv), or any other antibody that comprises, for example, a single
polypeptide chain comprising all six CDRs (three heavy chain CDRs
and three light chain CDRs) is considered to have a heavy chain and
a light chain. In some such embodiments, the heavy chain is the
region of the antibody that comprises the three heavy chain CDRs
and the light chain in the region of the antibody that comprises
the three light chain CDRs.
[0065] The term "heavy chain variable region" as used herein refers
to a region comprising heavy chain CDR1, framework (FR) 2, CDR2,
FR3, and CDR3. In some embodiments, a heavy chain variable region
also comprises at least a portion of an FR1 and/or at least a
portion of an FR4. In some embodiments, a heavy chain CDR1
corresponds to Kabat residues 26 to 35; a heavy chain CDR2
corresponds to Kabat residues 50 to 65; and a heavy chain CDR3
corresponds to Kabat residues 95 to 102. See, e.g., Kabat Sequences
of Proteins of Immunological Interest (1987 and 1991, NIH,
Bethesda, Md.); and FIG. 1. In some embodiments, a heavy chain CDR1
corresponds to Kabat residues 31 to 35; a heavy chain CDR2
corresponds to Kabat residues 50 to 65; and a heavy chain CDR3
corresponds to Kabat residues 95 to 102. See id.
[0066] The term "heavy chain constant region" as used herein refers
to a region comprising at least three heavy chain constant domains,
C.sub.H1, C.sub.H2, and C.sub.H3. Nonlimiting exemplary heavy chain
constant regions include .gamma., .delta., and .alpha.. Nonlimiting
exemplary heavy chain constant regions also include .epsilon. and
.mu.. Each heavy constant region corresponds to an antibody
isotype. For example, an antibody comprising a .gamma. constant
region is an IgG antibody, an antibody comprising a .delta.
constant region is an IgD antibody, and an antibody comprising an
.alpha. constant region is an IgA antibody. Further, an antibody
comprising a .mu. constant region is an IgM antibody, and an
antibody comprising an c constant region is an IgE antibody.
Certain isotypes can be further subdivided into subclasses. For
example, IgG antibodies include, but are not limited to, IgG1
(comprising a .gamma..sub.1 constant region), IgG2 (comprising a
.gamma..sub.2 constant region), IgG3 (comprising a .gamma..sub.3
constant region), and IgG4 (comprising a .gamma..sub.4 constant
region) antibodies; IgA antibodies include, but are not limited to,
IgA1 (comprising an .alpha..sub.1 constant region) and IgA2
(comprising an .alpha..sub.2 constant region) antibodies; and IgM
antibodies include, but are not limited to, IgM1 and IgM2.
[0067] In some embodiments, a heavy chain constant region comprises
one or more mutations (or substitutions), additions, or deletions
that confer a desired characteristic on the antibody. A nonlimiting
exemplary mutation is the S241P mutation in the IgG4 hinge region
(between constant domains C.sub.H1 and C.sub.H2), which alters the
IgG4 motif CPSCP to CPPCP, which is similar to the corresponding
motif in IgG1. That mutation, in some embodiments, results in a
more stable IgG4 antibody. See, e.g., Angal et al., Mol. Immunol.
30: 105-108 (1993); Bloom et al., Prot. Sci. 6: 407-415 (1997);
Schuurman et al., Mol. Immunol. 38: 1-8 (2001).
[0068] The term "heavy chain" as used herein refers to a
polypeptide comprising at least a heavy chain variable region, with
or without a leader sequence. In some embodiments, a heavy chain
comprises at least a portion of a heavy chain constant region. The
term "full-length heavy chain" as used herein refers to a
polypeptide comprising a heavy chain variable region and a heavy
chain constant region, with or without a leader sequence.
[0069] The term "light chain variable region" as used herein refers
to a region comprising light chain CDR1, framework (FR) 2, CDR2,
FR3, and CDR3. In some embodiments, a light chain variable region
also comprises an FR1 and/or an FR4. In some embodiments, a light
chain CDR1 corresponds to Kabat residues 24 to 34; a light chain
CDR2 corresponds to Kabat residues 50 to 56; and a light chain CDR3
corresponds to Kabat residues 89 to 97. See, e.g., Kabat Sequences
of Proteins of Immunological Interest (1987 and 1991, NIH,
Bethesda, Md.); and FIG. 1.
[0070] The term "light chain constant region" as used herein refers
to a region comprising a light chain constant domain, C.sub.L.
Nonlimiting exemplary light chain constant regions include .lamda.
and .kappa..
[0071] The term "light chain" as used herein refers to a
polypeptide comprising at least a light chain variable region, with
or without a leader sequence. In some embodiments, a light chain
comprises at least a portion of a light chain constant region. The
term "full-length light chain" as used herein refers to a
polypeptide comprising a light chain variable region and a light
chain constant region, with or without a leader sequence.
[0072] A "chimeric antibody" as used herein refers to an antibody
comprising at least one variable region from a first species (such
as mouse, rat, cynomolgus monkey, etc.) and at least one constant
region from a second species (such as human, cynomolgus monkey,
etc.). In some embodiments, a chimeric antibody comprises at least
one mouse variable region and at least one human constant region.
In some embodiments, a chimeric antibody comprises at least one
cynomolgus variable region and at least one human constant region.
In some embodiments, a chimeric antibody comprises at least one rat
variable region and at least one mouse constant region. In some
embodiments, all of the variable regions of a chimeric antibody are
from a first species and all of the constant regions of the
chimeric antibody are from a second species.
[0073] A "humanized antibody" as used herein refers to an antibody
in which at least one amino acid in a framework region of a
non-human variable region has been replaced with the corresponding
amino acid from a human variable region. In some embodiments, a
humanized antibody comprises at least one human constant region or
fragment thereof. In some embodiments, a humanized antibody is an
Fab, an scFv, a (Fab').sub.2, etc.
[0074] A "CDR-grafted antibody" as used herein refers to a
humanized antibody in which the complementarity determining regions
(CDRs) of a first (non-human) species have been grafted onto the
framework regions (FRs) of a second (human) species.
[0075] A "human antibody" as used herein refers to antibodies
produced in humans, antibodies produced in non-human animals that
comprise human immunoglobulin genes, such as XenoMouse.RTM., and
antibodies selected using in vitro methods, such as phage display,
wherein the antibody repertoire is based on a human immunoglobulin
sequences.
[0076] The term "leader sequence" refers to a sequence of amino
acid residues located at the N terminus of a polypeptide that
facilitates secretion of a polypeptide from a mammalian cell. A
leader sequence may be cleaved upon export of the polypeptide from
the mammalian cell, forming a mature protein. Leader sequences may
be natural or synthetic, and they may be heterologous or homologous
to the protein to which they are attached. Exemplary leader
sequences include, but are not limited to, antibody leader
sequences, such as, for example, the amino acid sequences of SEQ ID
NOs: 3 and 4, which correspond to human light and heavy chain
leader sequences, respectively. Nonlimiting exemplary leader
sequences also include leader sequences from heterologous proteins.
In some embodiments, an antibody lacks a leader sequence. In some
embodiments, an antibody comprises at least one leader sequence,
which may be selected from native antibody leader sequences and
heterologous leader sequences.
[0077] The term "vector" is used to describe a polynucleotide that
may be engineered to contain a cloned polynucleotide or
polynucleotides that may be propagated in a host cell. A vector may
include one or more of the following elements: an origin of
replication, one or more regulatory sequences (such as, for
example, promoters and/or enhancers) that regulate the expression
of the polypeptide of interest, and/or one or more selectable
marker genes (such as, for example, antibiotic resistance genes and
genes that may be used in colorimetric assays, e.g.,
.beta.-galactosidase). The term "expression vector" refers to a
vector that is used to express a polypeptide of interest in a host
cell.
[0078] A "host cell" refers to a cell that may be or has been a
recipient of a vector or isolated polynucleotide. Host cells may be
prokaryotic cells or eukaryotic cells. Exemplary eukaryotic cells
include mammalian cells, such as primate or non-primate animal
cells; fungal cells, such as yeast; plant cells; and insect cells.
Nonlimiting exemplary mammalian cells include, but are not limited
to, NSO cells, PER.C6.RTM. cells (Crucell), and 293 and CHO cells,
and their derivatives, such as 293-6E and DG44 cells,
respectively.
[0079] The term "isolated" as used herein refers to a molecule that
has been separated from at least some of the components with which
it is typically found in nature. For example, a polypeptide is
referred to as "isolated" when it is separated from at least some
of the components of the cell in which it was produced. Where a
polypeptide is secreted by a cell after expression, physically
separating the supernatant containing the polypeptide from the cell
that produced it is considered to be "isolating" the polypeptide.
Similarly, a polynucleotide is referred to as "isolated" when it is
not part of the larger polynucleotide (such as, for example,
genomic DNA or mitochondrial DNA, in the case of a DNA
polynucleotide) in which it is typically found in nature, or is
separated from at least some of the components of the cell in which
it was produced, e.g., in the case of an RNA polynucleotide. Thus,
a DNA polynucleotide that is contained in a vector inside a host
cell may be referred to as "isolated" so long as that
polynucleotide is not found in that vector in nature.
[0080] The terms "subject" and "patient" are used interchangeably
herein to refer to a human. In some embodiments, methods of
treating other mammals, including, but not limited to, rodents,
simians, felines, canines, equines, bovines, porcines, ovines,
caprines, mammalian laboratory animals, mammalian farm animals,
mammalian sport animals, and mammalian pets, are also provided.
[0081] As used herein, "rheumatoid arthritis" or "RA" refers to a
recognized disease state that may be diagnosed according to the
2000 revised American Rheumatoid Association criteria for the
classification of RA, or any similar criteria. In some embodiments,
the term "rheumatoid arthritis" refers to a chronic autoimmune
disease characterized primarily by inflammation of the lining
(synovium) of the joints, which can lead to joint damage, resulting
in chronic pain, loss of function, and disability. Because RA can
affect multiple organs of the body, including skin, lungs, and
eyes, it is referred to as a systemic illness.
[0082] The term "rheumatoid arthritis" includes not only active and
early RA, but also incipient RA, as defined below. Physiological
indicators of RA include, symmetric joint swelling which is
characteristic though not invariable in RA. Fusiform swelling of
the proximal interphalangeal (PIP) joints of the hands as well as
metacarpophalangeal (MCP), wrists, elbows, knees, ankles, and
metatarsophalangeal (MTP) joints are commonly affected and swelling
is easily detected. Pain on passive motion is the most sensitive
test for joint inflammation, and inflammation and structural
deformity often limits the range of motion for the affected joint.
Typical visible changes include ulnar deviation of the fingers at
the MCP joints, hyperextension, or hyperflexion of the MCP and PIP
joints, flexion contractures of the elbows, and subluxation of the
carpal bones and toes. The subject with RA may be resistant to a
disease-modifying anti-rheumatic drug (DMARD), and/or a
non-steroidal anti-inflammatory drug (NSAID). Nonlimiting exemplary
"DMARDs" include hydroxycloroquine, sulfasalazine, methotrexate
(MTX), leflunomide, etanercept, infliximab (plus oral and
subcutaneous MTX), azathioprine, D-penicillamine, gold salts
(oral), gold salts (intramuscular), minocycline, cyclosporine
including cyclosporine A and topical cyclosporine, staphylococcal
protein A (Goodyear and Silverman, J. Exp. Med., 197(9):1125-1139
(2003)), including salts and derivatives thereof, etc. Further
candidates for therapy according to this invention include those
who have experienced an inadequate response to previous or current
treatment with TNF inhibitors such as etanercept, infliximab and/or
adalimumab because of toxicity or inadequate efficacy.
[0083] A patient with "active rheumatoid arthritis" means a patient
with active and not latent symptoms of RA. Subjects with "early
active rheumatoid arthritis" are those subjects with active RA
diagnosed for at least 8 weeks but no longer than four years,
according to the revised 1987 ACR criteria for the classification
of RA.
[0084] Subjects with "early rheumatoid arthritis" are those
subjects with RA diagnosed for at least eight weeks but no longer
than four years, according to the revised 1987 ACR criteria for
classification of RA. RA includes, for example, juvenile-onset RA,
juvenile idiopathic arthritis (JIA), or juvenile RA (JRA).
[0085] Patients with "incipient RA" have early polyarthritis that
does not fully meet ACR criteria for a diagnosis of RA, in
association with the presence of RA-specific prognostic biomarkers
such as anti-CCP and shared epitope. They include patients with
positive anti-CCP antibodies who present with polyarthritis, but do
not yet have a diagnosis of RA, and are at high risk for going on
to develop bona fide ACR criteria RA (95% probability).
[0086] "Joint damage" is used in the broadest sense and refers to
damage or partial or complete destruction to any part of one or
more joints, including the connective tissue and cartilage, where
damage includes structural and/or functional damage of any cause,
and may or may not cause joint pain/arthalgia. It includes, without
limitation, joint damage associated with or resulting from
inflammatory joint disease as well as non-inflammatory joint
disease. This damage may be caused by any condition, such as an
autoimmune disease, especially arthritis, and most especially RA.
Exemplary such conditions include acute and chronic arthritis,
rheumatoid arthritis (including juvenile-onset RA, juvenile
idiopathic arthritis (JIA), and juvenile rheumatoid arthritis
(JRA)), and stages such as rheumatoid synovitis, gout or gouty
arthritis, acute immunological arthritis, chronic inflammatory
arthritis, degenerative arthritis, type II collagen-induced
arthritis, infectious arthritis, septic arthritis, Lyme arthritis,
proliferative arthritis, psoriatic arthritis, Still's disease,
vertebral arthritis, osteoarthritis, arthritis chronica
progrediente, arthritis deformans, polyarthritis chronica primaria,
reactive arthritis, menopausal arthritis, estrogen-depletion
arthritis, and ankylosing spondylitis/rheumatoid spondylitis),
rheumatic autoimmune disease other than RA, and significant
systemic involvement secondary to RA (including but not limited to
vasculitis, pulmonary fibrosis or Felty's syndrome). For purposes
herein, joints are points of contact between elements of a skeleton
(of a vertebrate such as an animal) with the parts that surround
and support it and include, but are not limited to, for example,
hips, joints between the vertebrae of the spine, joints between the
spine and pelvis (sacroiliac joints), joints where the tendons and
ligaments attach to bones, joints between the ribs and spine,
shoulders, knees, feet, elbows, hands, fingers, ankles and toes,
but especially joints in the hands and feet.
[0087] The term "lupus" as used herein is an autoimmune disease or
disorder that in general involves antibodies that attack connective
tissue. The principal form of lupus is a systemic one, systemic
lupus erythematosus (SLE), including cutaneous SLE and
subacutecutaneous SLE, as well as other types of lupus (including
nephritis, extrarenal, cerebritis, pediatric, non-renal, discoid,
and alopecia). In certain embodiments, the term "systemic lupus
erythematosus" refers to a chronic autoimmune disease that can
result in skin lesions, joint pain and swelling, kidney disease
(lupus nephritis), fluid around the heart and/or lungs,
inflammation of the heart, and various other systemic conditions.
In certain embodiments, the term "lupus nephritis" refers to
inflammation of the kidneys that occurs in patients with SLE. Lupus
nephritis may include, for example, glomerulonephritis and/or
interstitial nephritis, and can lead to hypertension, proteinuria,
and kidney failure. Lupus nephritis may be classified based on
severity and extent of disease, for example, as defined by the
International Society of Nephrology/Renal/Pathology Society. Lupus
nephritis classes include class I (minimal mesangial lupus
nephritis), class II (mesangial proliferative lupus nephritis),
class III (focal lupus nephritis), class IV (diffuse segmental
(IV-S) or diffuse global (IV-G) lupus nephritis), class V
(membranous lupus nephritis), and class VI (advanced sclerosing
lupus nephritis). The term "lupus nephritis" encompasses all of the
classes.
[0088] The term "multiple sclerosis" ("MS") refers to the chronic
and often disabling disease of the central nervous system
characterized by the progressive destruction of the myelin.
"Demyelination" occurs when the myelin sheath becomes inflamed,
injured, and detaches from the nerve fiber. There are four
internationally recognized forms of MS, namely, primary progressive
multiple sclerosis (PPMS), relapsing-remitting multiple sclerosis
(RRMS), secondary progressive multiple sclerosis (SPMS), and
progressive relapsing multiple sclerosis (PRMS).
[0089] "Primary progressive multiple sclerosis" or "PPMS" is
characterized by a gradual progression of the disease from its
onset with no superimposed relapses and remissions at all. There
may be periods of a leveling off of disease activity and there may
be good and bad days or weeks. PPMS differs from RRMS and SPMS in
that onset is typically in the late thirties or early forties, men
are as likely as women to develop it, and initial disease activity
is often in the spinal cord and not in the brain. PPMS often
migrates into the brain, but is less likely to damage brain areas
than RRMS or SPMS; for example, people with PPMS are less likely to
develop cognitive problems. PPMS is the sub-type of MS that is
least likely to show inflammatory (gadolinium enhancing) lesions on
MRI scans. The primary progressive form of the disease affects
between 10 and 15% of all people with multiple sclerosis. PPMS may
be defined according to the criteria in McDonald et al. Ann Neurol
50:121-7 (2001). The subject with PPMS treated herein is usually
one with a probable or definitive diagnosis of PPMS.
[0090] "Relapsing-remitting multiple sclerosis" or "RRMS" is
characterized by relapses (also known as exacerbations) during
which time new symptoms can appear and old ones resurface or
worsen. The relapses are followed by periods of remission, during
which time the person fully or partially recovers from the deficits
acquired during the relapse. Relapses can last for days, weeks, or
months, and recovery can be slow and gradual or almost
instantaneous. The vast majority of people presenting with MS are
first diagnosed with RRMS. This is typically when they are in their
twenties or thirties, though diagnoses much earlier or later are
known. Twice as many women as men present with this sub-type of MS.
During relapses, myelin, a protective insulating sheath around the
nerve fibers (neurons) in the white matter regions of the central
nervous system (CNS), may be damaged in an inflammatory response by
the body's own immune system. This causes a wide variety of
neurological symptoms that vary considerably depending on which
areas of the CNS are damaged. Immediately after a relapse, the
inflammatory response dies down and a special type of glial cell in
the CNS (called an oligodendrocyte) sponsors remyelination--a
process whereby the myelin sheath around the axon may be repaired.
It is this remyelination that may be responsible for the remission.
Approximately 50% of patients with RRMS convert to SPMS within 10
years of disease onset. After 30 years, this figure rises to 90%.
At any one time, the relapsing-remitting form of the disease
accounts around 55% of all people with MS.
[0091] "Secondary progressive multiple sclerosis" or "SPMS" is
characterized by a steady progression of clinical neurological
damage with or without superimposed relapses and minor remissions
and plateau. People who develop SPMS will have previously
experienced a period of RRMS which may have lasted anywhere from
two to forty years or more. Any superimposed relapses and
remissions tend to tail off over time. From the onset of the
secondary progressive phase of the disease, disability starts
advancing much quicker than it did during RRMS though the progress
can still be quite slow in some individuals. SPMS tends to be
associated with lower levels of inflammatory lesion formation than
in RRMS but the total burden of disease continues to progress. At
any one time, SPMS accounts around 30% of all people with multiple
sclerosis.
[0092] "Progressive relapsing multiple sclerosis" or "PRMS" is
characterized by a steady progression of clinical neurological
damage with superimposed relapses and remissions. There is
significant recovery immediately following a relapse but between
relapses there is a gradual worsening of symptoms. PRMS affects
around 5% of all people with multiple sclerosis. Some neurologists
believe PRMS is a variant of PPMS.
[0093] The term "CD16+ disorder" means a disease in which CD16+
monocytes of a mammal cause, mediate or otherwise contribute to a
morbidity in the mammal. Also included are diseases in which
reduction of CD16+ monocytes has an ameliorative effect on
progression of the disease. Included within this term are CD16+
inflammatory diseases, infectious diseases, immunodeficiency
diseases, neoplasia, etc. In certain embodiments, CD16+
inflammatory diseases include inflammatory diseases that are not
responsive to methotrexate therapy. In certain embodiments, CD16+
inflammatory diseases include methotrexate-resistant rheumatoid
arthritis, methotrexate-resistant multiple sclerosis,
methotrexate-resistant lupus, methotrexate-resistant inflammatory
bowel disease, methotrexate-resistant Crohn's disease,
methotrexate-resistant asthma, and methotrexate-resistant
psoriasis. In certain embodiments, patients having
methotrexate-resistant diseases, such as methotrexate-resistant
rheumatoid arthritis, are referred to as methotrexate incomplete
responders or methotrexate inadequate responders.
[0094] Examples of CD16+ disorders that can be treated according to
the invention include, but are not limited to, systemic lupus
erythematosus, rheumatoid arthritis, juvenile chronic arthritis,
spondyloarthropathies, systemic sclerosis (scieroderma), idiopathic
inflammatory myopathies (dermatomyositis, polymyositis), Sjogren's
syndrome, systemic vasculitis, sarcoidosis, autoimmune hemolytic
anemia (immune pancytopenia, paroxysmal nocturnal hemoglobinuria),
autoimmune thrombocytopenia (idiopathic thrombocytopenic purpura,
immune-mediated thrombocytopenia), thyroiditis (Grave's disease,
Hashimoto's thyroiditis, juvenile lymphocytic thyroiditis, atrophic
thyroiditis), diabetes mellitus, immune-mediated renal disease
(glomerulonephritis, tubulointerstitial nephritis), demyelinating
diseases of the central and peripheral nervous systems such as
multiple sclerosis, idiopathic demyelinating polyneuropathy or
Guillain-Barre syndrome, and chronic inflammatory demyelinating
polyneuropathy, hepatobiliary diseases such as infectious hepatitis
(hepatitis A, B, C, D, E and other non-hepatotropic viruses),
autoimmune chronic active hepatitis, primary biliary cirrhosis,
granulomatous hepatitis, and sclerosing cholangitis, inflammatory
bowel disease (IBD), including ulcerative colitis, Crohn's disease,
gluten-sensitive enteropathy, and Whipple's disease, autoimmune or
immune-mediated skin diseases including bullous skin diseases,
erythema multiforme and contact dermatitis, psoriasis, allergic
diseases such as asthma, allergic rhinitis, atopic dermatitis, food
hypersensitivity and urticaria, immunologic diseases of the lung
such as eosinophilic pneumonia, idiopathic pulmonary fibrosis and
hypersensitivity pneumonitis, transplantation associated diseases
including graft rejection and graft-versus-host-disease; fibrosis,
including kidney fibrosis and hepatic fibrosis, cardiovascular
disease, including atherosclerosis and coronary artery disease,
cardiovascular events associated with chronic kidney disease,
myocardial infarction, and congestive heart failure, diabetes,
including type II diabetes, Bronchiolitis obliterans with
organizing pneumonia (BOOP), hemophagocytic syndrome, macrophage
activation syndrome, sarcoidosis, and periodontitis. Infectious
diseases including viral diseases such as AIDS (HIV infection),
hepatitis A, B, C, D, and E, herpes, etc., bacterial infections,
fungal infections, protozoal infections and parasitic
infections.
[0095] "Treatment," as used herein, refers to both therapeutic
treatment and prophylactic or preventative measures, wherein the
object is to prevent or slow down (lessen) the targeted pathologic
condition or disorder. In certain embodiments, the term "treatment"
covers any administration or application of a therapeutic for
disease in a mammal, including a human, and includes inhibiting or
slowing the disease or progression of the disease; partially or
fully relieving the disease, for example, by causing regression, or
restoring or repairing a lost, missing, or defective function;
stimulating an inefficient process; or causing the disease plateau
to have reduced severity. The term "treatment" also includes
reducing the severity of any phenotypic characteristic and/or
reducing the incidence, degree, or likelihood of that
characteristic. Those in need of treatment include those already
with the disorder as well as those prone to have the disorder or
those in whom the disorder is to be prevented.
[0096] "Chronic" administration refers to administration of an
agent in a continuous mode as opposed to an acute mode, so as to
maintain the initial therapeutic effect (activity) for an extended
period of time. "Intermittent" administration is treatment that is
not consecutively done without interruption, but rather is cyclic
in nature.
[0097] The term "effective amount" or "therapeutically effective
amount" refers to an amount of a drug effective to treat a disease
or disorder in a subject. In certain embodiments, an effective
amount refers to an amount effective, at dosages and for periods of
time necessary, to achieve the desired therapeutic or prophylactic
result. A therapeutically effective amount of an anti-CSF1R
antibody of the invention may vary according to factors such as the
disease state, age, sex, and weight of the individual, and the
ability of the anti-CSF1R antibody to elicit a desired response in
the individual. A therapeutically effective amount encompasses an
amount in which any toxic or detrimental effects of the anti-CSF1R
antibody are outweighed by the therapeutically beneficial
effects.
[0098] A "prophylactically effective amount" refers to an amount
effective, at dosages and for periods of time necessary, to achieve
the desired prophylactic result. Typically, but not necessarily,
since a prophylactic dose is used in subjects prior to or at an
earlier stage of disease, the prophylactically effective amount
would be less than the therapeutically effective amount.
[0099] Administration "in combination with" one or more further
therapeutic agents includes simultaneous (concurrent) and
consecutive administration in any order.
[0100] A "pharmaceutically acceptable carrier" refers to a
non-toxic solid, semisolid, or liquid filler, diluent,
encapsulating material, formulation auxiliary, or carrier
conventional in the art for use with a therapeutic agent that
together comprise a "pharmaceutical composition" for administration
to a subject. A pharmaceutically acceptable carrier is non-toxic to
recipients at the dosages and concentrations employed and is
compatible with other ingredients of the formulation. The
pharmaceutically acceptable carrier is appropriate for the
formulation employed. For example, if the therapeutic agent is to
be administered orally, the carrier may be a gel capsule. If the
therapeutic agent is to be administered subcutaneously, the carrier
ideally is not irritable to the skin and does not cause injection
site reaction.
Anti-CSF1R Antibodies
[0101] Anti-CSF1R antibodies include, but are not limited to,
humanized antibodies, chimeric antibodies, mouse antibodies, human
antibodies, and antibodies comprising the heavy chain and/or light
chain CDRs discussed herein.
[0102] Exemplary Humanized Antibodies
[0103] In some embodiments, humanized antibodies that bind CSF1R
are provided. Humanized antibodies are useful as therapeutic
molecules because humanized antibodies reduce or eliminate the
human immune response to non-human antibodies (such as the human
anti-mouse antibody (HAMA) response), which can result in an immune
response to an antibody therapeutic, and decreased effectiveness of
the therapeutic.
[0104] Nonlimiting exemplary humanized antibodies include huAb1
through huAb16, described herein. Nonlimiting exemplary humanized
antibodies also include antibodies comprising a heavy chain
variable region of an antibody selected from huAb1 to huAb16 and/or
a light chain variable region of an antibody selected from huAb1 to
huAb16. Nonlimiting exemplary humanized antibodies include
antibodies comprising a heavy chain variable region selected from
SEQ ID NOs: 39 to 45 and/or a light chain variable region selected
from SEQ ID NOs: 46 to 52. Exemplary humanized antibodies also
include, but are not limited to, humanized antibodies comprising
heavy chain CDR1, CDR2, and CDR3, and/or light chain CDR1, CDR2,
and CDR3 of an antibody selected from 0301, 0302, and 0311.
[0105] In some embodiments, a humanized anti-CSF1R antibody
comprises heavy chain CDR1, CDR2, and CDR3 and/or a light chain
CDR1, CDR2, and CDR3 of an antibody selected from 0301, 0302, and
0311. Nonlimiting exemplary humanized anti-CSF1R antibodies include
antibodies comprising sets of heavy chain CDR1, CDR2, and CDR3
selected from: SEQ ID NOs: 15, 16, and 17; SEQ ID NOs: 21, 22, and
23; and SEQ ID NOs: 27, 28, and 29. Nonlimiting exemplary humanized
anti-CSF1R antibodies also include antibodies comprising sets of
light chain CDR1, CDR2, and CDR3 selected from: SEQ ID NOs: 18, 19,
and 20; SEQ ID NOs: 24, 25, and 26; and SEQ ID NOs: 30, 31, and
32.
[0106] Nonlimiting exemplary humanized anti-CSF1R antibodies
include antibodies comprising the sets of heavy chain CDR1, CDR2,
and CDR3, and light chain CDR1, CDR2, and CDR3 in Table 1 (SEQ ID
NOs shown; see Table 8 for sequences). Each row of Table 1 shows
the heavy chain CDR1, CDR2, and CDR3, and light chain CDR1, CDR2,
and CDR3 of an exemplary antibody.
TABLE-US-00001 TABLE 1 Heavy chain and light chain CDRs Heavy chain
Light chain CDR1 CDR2 CDR3 CDR1 CDR2 CDR3 Ab SEQ ID SEQ ID SEQ ID
SEQ ID SEQ ID SEQ ID 0301 15 16 17 18 19 20 0302 21 22 23 24 25 26
0311 27 28 29 30 31 32
[0107] Further Exemplary Humanized Antibodies
[0108] In some embodiments, a humanized anti-CSF1R antibody
comprises a heavy chain comprising a variable region sequence that
is 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% identical to a sequence selected from SEQ ID NOs: 9, 11,
13, and 39 to 45, and wherein the antibody binds CSF1R. In some
embodiments, a humanized anti-CSF1R antibody comprises a light
chain comprising a variable region sequence that is 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%
identical to a sequence selected from SEQ ID NOs: 10, 12, 14, and
46 to 52, wherein the antibody binds CSF1R. In some embodiments, a
humanized anti-CSF1R antibody comprises a heavy chain comprising a
variable region sequence that is 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% identical to a sequence
selected from SEQ ID NOs: 9, 11, 13, and 39 to 45; and a light
chain comprising a variable region sequence that is 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%
identical to a sequence selected from SEQ ID NOs: 10, 12, 14, and
46 to 52; wherein the antibody binds CSF1R.
[0109] As used herein, whether a particular polypeptide is, for
example, at least 95% identical to an amino acid sequence can be
determined using, e.g., a computer program. When determining
whether a particular sequence is, for example, 95% identical to a
reference sequence, the percentage of identity is calculated over
the full length of the reference amino acid sequence.
[0110] In some embodiments, a humanized anti-CSF1R antibody
comprises at least one of the CDRs discussed herein. That is, in
some embodiments, a humanized anti-CSF1R antibody comprises at
least one CDR selected from a heavy chain CDR1 discussed herein, a
heavy chain CDR2 discussed herein, a heavy chain CDR3 discussed
herein, a light chain CDR1 discussed herein, a light chain CDR2
discussed herein, and a light chain CDR3 discussed herein. Further,
in some embodiments, a humanized anti-CSF1R antibody comprises at
least one mutated CDR based on a CDR discussed herein, wherein the
mutated CDR comprises 1, 2, 3, or 4 amino acid substitutions
relative to the CDR discussed herein. In some embodiments, one or
more of the amino acid substitutions are conservative amino acid
substitutions. One skilled in the art can select one or more
suitable conservative amino acid substitutions for a particular CDR
sequence, wherein the suitable conservative amino acid
substitutions are not predicted to significantly alter the binding
properties of the antibody comprising the mutated CDR.
[0111] Exemplary humanized anti-CSF1R antibodies also include
antibodies that compete for binding to CSF1R with an antibody
described herein. Thus, in some embodiments, a humanized anti-CSF1R
antibody is provided that competes for binding to CSF1R with an
antibody selected from Fabs 0301, 0302, and 0311; and bivalent
(i.e., having two heavy chains and two light chains) antibody
versions of those Fabs.
[0112] Exemplary Humanized Antibody Constant Regions
[0113] In some embodiments, a humanized antibody described herein
comprises one or more human constant regions. In some embodiments,
the human heavy chain constant region is of an isotype selected
from IgA, IgG, and IgD. In some embodiments, the human light chain
constant region is of an isotype selected from .kappa. and .lamda..
In some embodiments, a humanized antibody described herein
comprises a human IgG constant region. In some embodiments, a
humanized antibody described herein comprises a human IgG4 heavy
chain constant region. In some such embodiments, a humanized
antibody described herein comprises an S241P mutation in the human
IgG4 constant region. In some embodiments, a humanized antibody
described herein comprises a human IgG4 constant region and a human
.kappa. light chain.
[0114] The choice of heavy chain constant region can determine
whether or not an antibody will have effector function in vivo.
Such effector function, in some embodiments, includes
antibody-dependent cell-mediated cytotoxicity (ADCC) and/or
complement-dependent cytotoxicity (CDC), and can result in killing
of the cell to which the antibody is bound. In some methods of
treatment, including methods of treating some cancers, cell killing
may be desirable, for example, when the antibody binds to a cell
that supports the maintenance or growth of the tumor. Exemplary
cells that may support the maintenance or growth of a tumor
include, but are not limited to, tumor cells themselves, cells that
aid in the recruitment of vasculature to the tumor, and cells that
provide ligands, growth factors, or counter-receptors that support
or promote tumor growth or tumor survival. In some embodiments,
when effector function is desirable, an anti-CSF1R antibody
comprising a human IgG1 heavy chain or a human IgG3 heavy chain is
selected.
[0115] In some methods of treatment, effector function may not be
desirable. For example, in some embodiments, it may be desirable
that antibodies used in the treatment of lupus and/or MS and/or RA
and/or osteolysis do not have effector function. Thus, in some
embodiments, anti-CSF1R antibodies developed for the treatment of
cancer may not be suitable for use in treatment of lupus and/or MS
and/or RA and/or osteolysis. Accordingly, in some embodiments, an
anti-CSF1R antibody that lacks significant effector function is
used in treatment of lupus and/or MS and/or RA and/or osteolysis.
In some embodiments, an anti-CSF1R antibody for treatment of lupus
and/or MS and/or RA and/or osteolysis comprises a human IgG4 or
IgG2 heavy chain constant region. In some embodiments, an IgG4
constant region comprises an S241P mutation.
[0116] An antibody may be humanized by any method. Nonlimiting
exemplary methods of humanization include methods described, e.g.,
in U.S. Pat. Nos. 5,530,101; 5,585,089; 5,693,761; 5,693,762;
6,180,370; Jones et al., Nature 321: 522-525 (1986); Riechmann et
al., Nature 332: 323-27 (1988); Verhoeyen et al., Science 239:
1534-36 (1988); and U.S. Publication No. US 2009/0136500.
[0117] As noted above, a humanized antibody is an antibody in which
at least one amino acid in a framework region of a non-human
variable region has been replaced with the amino acid from the
corresponding location in a human framework region. In some
embodiments, at least two, at least three, at least four, at least
five, at least six, at least seven, at least eight, at least nine,
at least 10, at least 11, at least 12, at least 15, or at least 20
amino acids in the framework regions of a non-human variable region
are replaced with an amino acid from one or more corresponding
locations in one or more human framework regions.
[0118] In some embodiments, some of the corresponding human amino
acids used for substitution are from the framework regions of
different human immunoglobulin genes. That is, in some such
embodiments, one or more of the non-human amino acids may be
replaced with corresponding amino acids from a human framework
region of a first human antibody or encoded by a first human
immunoglobulin gene, one or more of the non-human amino acids may
be replaced with corresponding amino acids from a human framework
region of a second human antibody or encoded by a second human
immunoglobulin gene, one or more of the non-human amino acids may
be replaced with corresponding amino acids from a human framework
region of a third human antibody or encoded by a third human
immunoglobulin gene, etc. Further, in some embodiments, all of the
corresponding human amino acids being used for substitution in a
single framework region, for example, FR2, need not be from the
same human framework. In some embodiments, however, all of the
corresponding human amino acids being used for substitution are
from the same human antibody or encoded by the same human
immunoglobulin gene.
[0119] In some embodiments, an antibody is humanized by replacing
one or more entire framework regions with corresponding human
framework regions. In some embodiments, a human framework region is
selected that has the highest level of homology to the non-human
framework region being replaced. In some embodiments, such a
humanized antibody is a CDR-grafted antibody.
[0120] In some embodiments, following CDR-grafting, one or more
framework amino acids are changed back to the corresponding amino
acid in a mouse framework region. Such "back mutations" are made,
in some embodiments, to retain one or more mouse framework amino
acids that appear to contribute to the structure of one or more of
the CDRs and/or that may be involved in antigen contacts and/or
appear to be involved in the overall structural integrity of the
antibody. In some embodiments, ten or fewer, nine or fewer, eight
or fewer, seven or fewer, six or fewer, five or fewer, four or
fewer, three or fewer, two or fewer, one, or zero back mutations
are made to the framework regions of an antibody following CDR
grafting.
[0121] In some embodiments, a humanized antibody also comprises a
human heavy chain constant region and/or a human light chain
constant region.
[0122] Exemplary Chimeric Antibodies
[0123] In some embodiments, an anti-CSF1R antibody is a chimeric
antibody. In some embodiments, an anti-CSF1R antibody comprises at
least one non-human variable region and at least one human constant
region. In some such embodiments, all of the variable regions of an
anti-CSF1R antibody are non-human variable regions, and all of the
constant regions of an anti-CSF1R antibody are human constant
regions. In some embodiments, one or more variable regions of a
chimeric antibody are mouse variable regions. The human constant
region of a chimeric antibody need not be of the same isotype as
the non-human constant region, if any, it replaces. Chimeric
antibodies are discussed, e.g., in U.S. Pat. No. 4,816,567; and
Morrison et al. Proc. Natl. Acad. Sci. USA 81: 6851-55 (1984).
[0124] Nonlimiting exemplary chimeric antibodies include chimeric
antibodies comprising the heavy and/or light chain variable regions
of an antibody selected from 0301, 0302, and 0311. Additional
nonlimiting exemplary chimeric antibodies include chimeric
antibodies comprising heavy chain CDR1, CDR2, and CDR3, and/or
light chain CDR1, CDR2, and CDR3 of an antibody selected from 0301,
0302, and 0311.
[0125] Nonlimiting exemplary chimeric anti-CSF1R antibodies include
antibodies comprising the following pairs of heavy and light chain
variable regions: SEQ ID NOs: 9 and 10; SEQ ID NOs: 11 and 12; and
SEQ ID NOs: 13 and 14.
[0126] Nonlimiting exemplary anti-CSF1R antibodies include
antibodies comprising a set of heavy chain CDR1, CDR2, and CDR3,
and light chain CDR1, CDR2, and CDR3 shown above in Table 1.
[0127] Further Exemplary Chimeric Antibodies
[0128] In some embodiments, a chimeric anti-CSF1R antibody
comprises a heavy chain comprising a variable region sequence that
is 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% identical to a sequence selected from SEQ ID NOs: 9, 11,
13, and 39 to 45, wherein the antibody binds CSF1R. In some
embodiments, a chimeric anti-CSF1R antibody comprises a light chain
comprising a variable region sequence that is 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% identical
to a sequence selected from SEQ ID NOs: 10, 12, 14, and 46 to 52,
wherein the antibody binds CSF1R. In some embodiments, a chimeric
anti-CSF1R antibody comprises a heavy chain comprising a variable
region sequence that is 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% identical to a sequence selected
from SEQ ID NOs: 9, 11, 13, and 39 to 45; and a light chain
comprising a variable region sequence that is 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% identical
to a sequence selected from SEQ ID NOs: 10, 12, 14, and 46 to 52;
wherein the antibody binds CSF1R.
[0129] In some embodiments, a chimeric anti-CSF1R antibody
comprises at least one of the CDRs discussed herein. That is, in
some embodiments, a chimeric anti-CSF1R antibody comprises at least
one CDR selected from a heavy chain CDR1 discussed herein, a heavy
chain CDR2 discussed herein, a heavy chain CDR3 discussed herein, a
light chain CDR1 discussed herein, a light chain CDR2 discussed
herein, and a light chain CDR3 discussed herein. Further, in some
embodiments, a chimeric anti-CSF1R antibody comprises at least one
mutated CDR based on a CDR discussed herein, wherein the mutated
CDR comprises 1, 2, 3, or 4 amino acid substitutions relative to
the CDR discussed herein. In some embodiments, one or more of the
amino acid substitutions are conservative amino acid substitutions.
One skilled in the art can select one or more suitable conservative
amino acid substitutions for a particular CDR sequence, wherein the
suitable conservative amino acid substitutions are not predicted to
significantly alter the binding properties of the antibody
comprising the mutated CDR.
[0130] Exemplary chimeric anti-CSF1R antibodies also include
chimeric antibodies that compete for binding to CSF1R with an
antibody described herein. Thus, in some embodiments, a chimeric
anti-CSF1R antibody is provided that competes for binding to CSF1R
with an antibody selected from Fabs 0301, 0302, and 0311; and
bivalent (i.e., having two heavy chains and two light chains)
antibody versions of those Fabs.
[0131] Exemplary Chimeric Antibody Constant Regions
[0132] In some embodiments, a chimeric antibody described herein
comprises one or more human constant regions. In some embodiments,
the human heavy chain constant region is of an isotype selected
from IgA, IgG, and IgD. In some embodiments, the human light chain
constant region is of an isotype selected from .kappa. and .lamda..
In some embodiments, a chimeric antibody described herein comprises
a human IgG constant region. In some embodiments, a chimeric
antibody described herein comprises a human IgG4 heavy chain
constant region. In some such embodiments, a chimeric antibody
described herein comprises an S241P mutation in the human IgG4
constant region. In some embodiments, a chimeric antibody described
herein comprises a human IgG4 constant region and a human .kappa.
light chain.
[0133] As noted above, whether or not effector function is
desirable may depend on the particular method of treatment intended
for an antibody. Thus, in some embodiments, when effector function
is desirable, a chimeric anti-CSF1R antibody comprising a human
IgG1 heavy chain constant region or a human IgG3 heavy chain
constant region is selected. In some embodiments, when effector
function is not desirable, a chimeric anti-CSF1R antibody
comprising a human IgG4 or IgG2 heavy chain constant region is
selected.
[0134] Exemplary Human Antibodies
[0135] Human antibodies can be made by any suitable method.
Nonlimiting exemplary methods include making human antibodies in
transgenic mice that comprise human immunoglobulin loci. See, e.g.,
Jakobovits et al., Proc. Natl. Acad. Sci. USA 90: 2551-55 (1993);
Jakobovits et al., Nature 362: 255-8 (1993); Lonberg et al., Nature
368: 856-9 (1994); and U.S. Pat. Nos. 5,545,807; 6,713,610;
6,673,986; 6,162,963; 5,545,807; 6,300,129; 6,255,458; 5,877,397;
5,874,299; and 5,545,806.
[0136] Nonlimiting exemplary methods also include making human
antibodies using phage display libraries. See, e.g., Hoogenboom et
al., J. Mol. Biol. 227: 381-8 (1992); Marks et al., J. Mol. Biol.
222: 581-97 (1991); and PCT Publication No. WO 99/10494.
[0137] In some embodiments, a human anti-CSF1R antibody binds to a
polypeptide having the sequence of SEQ ID NO: 1. Exemplary human
anti-CSF1R antibodies also include antibodies that compete for
binding to CSF1R with an antibody described herein. Thus, in some
embodiments, a human anti-CSF1R antibody is provided that competes
for binding to CSF1R with an antibody selected from Fabs 0301,
0302, and 0311, and bivalent (i.e., having two heavy chains and two
light chains) antibody versions of those Fabs.
[0138] In some embodiments, a human anti-CSF1R antibody comprises
one or more human constant regions. In some embodiments, the human
heavy chain constant region is of an isotype selected from IgA,
IgG, and IgD. In some embodiments, the human light chain constant
region is of an isotype selected from .kappa. and .lamda.. In some
embodiments, a human antibody described herein comprises a human
IgG constant region. In some embodiments, a human antibody
described herein comprises a human IgG4 heavy chain constant
region. In some such embodiments, a human antibody described herein
comprises an S241P mutation in the human IgG4 constant region. In
some embodiments, a human antibody described herein comprises a
human IgG4 constant region and a human .kappa. light chain.
[0139] In some embodiments, when effector function is desirable, a
human anti-CSF1R antibody comprising a human IgG1 heavy chain
constant region or a human IgG3 heavy chain constant region is
selected. In some embodiments, when effector function is not
desirable, a human anti-CSF1R antibody comprising a human IgG4 or
IgG2 heavy chain constant region is selected.
[0140] Additional Exemplary Anti-CSF1R Antibodies
[0141] Exemplary anti-CSF1R antibodies also include, but are not
limited to, mouse, humanized, human, chimeric, and engineered
antibodies that comprise, for example, one or more of the CDR
sequences described herein. In some embodiments, an anti-CSF1R
antibody comprises a heavy chain variable region described herein.
In some embodiments, an anti-CSF1R antibody comprises a light chain
variable region described herein. In some embodiments, an
anti-CSF1R antibody comprises a heavy chain variable region
described herein and a light chain variable region described
herein. In some embodiments, an anti-CSF1R antibody comprises heavy
chain CDR1, CDR2, and CDR3 described herein. In some embodiments,
an anti-CSF1R antibody comprises light chain CDR1, CDR2, and CDR3
described herein. In some embodiments, an anti-CSF1R antibody
comprises heavy chain CDR1, CDR2, and CDR3 described herein and
light chain CDR1, CDR2, and CDR3 described herein.
[0142] In some embodiments, an anti-CSF1R antibody comprises a
heavy chain variable region of an antibody selected from Fabs 0301,
0302, and 0311. Nonlimiting exemplary anti-CSF1R antibodies also
include antibodies comprising a heavy chain variable region of an
antibody selected from humanized antibodies huAb1 to huAb16.
Nonlimiting exemplary anti-CSF1R antibodies include antibodies
comprising a heavy chain variable region comprising a sequence
selected from SEQ ID NOs: 9, 11, 13, and 39 to 45.
[0143] In some embodiments, an anti-CSF1R antibody comprises a
light chain variable region of an antibody selected from Fabs 0301,
0302, and 311. Nonlimiting exemplary anti-CSF1R antibodies also
include antibodies comprising a light chain variable region of an
antibody selected from humanized antibodies huAb1 to huAb16.
Nonlimiting exemplary anti-CSF1R antibodies include antibodies
comprising a light chain variable region comprising a sequence
selected from SEQ ID NOs: 10, 12, 14, and 46 to 52.
[0144] In some embodiments, an anti-CSF1R antibody comprises a
heavy chain variable region and a light chain variable region of an
antibody selected from Fabs 0301, 0302, and 0311. Nonlimiting
exemplary anti-CSF1R antibodies also include antibodies comprising
a heavy chain variable region and a light chain variable region of
an antibody selected from humanized antibodies huAb1 to huAb16.
Nonlimiting exemplary anti-CSF1R antibodies include antibodies
comprising the following pairs of heavy and light chain variable
regions: SEQ ID NOs: 9 and 10; SEQ ID NOs: 11 and 12; and SEQ ID
NOs: 13 and 14; SEQ ID NOs: 39 and 40; SEQ ID NOs: 41 and 42; SEQ
ID NOs: 43 and 44; SEQ ID NOs: 45 and 46; SEQ ID NOs: 47 and 48;
SEQ ID NOs: 49 and 50; and SEQ ID NOs: 51 and 52. Nonlimiting
exemplary anti-CSF1R antibodies also include antibodies comprising
the following pairs of heavy and light chains: SEQ ID NOs: 33 and
34; SEQ ID NOs: 35 and 36; and SEQ ID NOs: 37 and 38.
[0145] In some embodiments, an anti-CSF1R antibody comprises heavy
chain CDR1, CDR2, and CDR3 of an antibody selected from Fabs 0301,
0302, and 0311. Nonlimiting exemplary anti-CSF1R antibodies include
antibodies comprising sets of heavy chain CDR1, CDR2, and CDR3
selected from: SEQ ID NOs: 15, 16, and 17; SEQ ID NOs: 21, 22, and
23; and SEQ ID NOs: 27, 28, and 29.
[0146] In some embodiments, an anti-CSF1R antibody comprises light
chain CDR1, CDR2, and CDR3 of an antibody selected from Fabs 0301,
0302, and 0311. Nonlimiting exemplary anti-CSF1R antibodies include
antibodies comprising sets of light chain CDR1, CDR2, and CDR3
selected from: SEQ ID NOs: 18, 19, and 20; SEQ ID NOs: 24, 25, and
26; and SEQ ID NOs: 30, 31, and 32.
[0147] In some embodiments, an anti-CSF1R antibody comprises heavy
chain CDR1, CDR2, and CDR3, and light chain CDR1, CDR2, and CDR3 of
an antibody selected from Fabs 0301, 0302, and 0311.
[0148] Nonlimiting exemplary anti-CSF1R antibodies include
antibodies comprising the sets of heavy chain CDR1, CDR2, and CDR3,
and light chain CDR1, CDR2, and CDR3 shown above in Table 1.
[0149] Further Exemplary Antibodies
[0150] In some embodiments, an anti-CSF1R antibody comprises a
heavy chain comprising a variable region sequence that is 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% identical to a sequence selected from SEQ ID NOs: 9, 11, 13,
and 39 to 45, wherein the antibody binds CSF1R. In some
embodiments, an anti-CSF1R antibody comprises a light chain
comprising a variable region sequence that is 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% identical
to a sequence selected from SEQ ID NOs: 10, 12, 14, and 46 to 52,
wherein the antibody binds CSF1R. In some embodiments, an
anti-CSF1R antibody comprises a heavy chain comprising a variable
region sequence that is 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% identical to a sequence selected
from SEQ ID NOs: 9, 11, 13, and 39 to 45; and a light chain
comprising a variable region sequence that is 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% identical
to a sequence selected from SEQ ID NOs: 10, 12, 14, and 46 to 52;
wherein the antibody binds CSF1R.
[0151] In some embodiments, an anti-CSF1R antibody comprises at
least one of the CDRs discussed herein. That is, in some
embodiments, an anti-CSF1R antibody comprises at least one CDR
selected from a heavy chain CDR1 discussed herein, a heavy chain
CDR2 discussed herein, a heavy chain CDR3 discussed herein, a light
chain CDR1 discussed herein, a light chain CDR2 discussed herein,
and a light chain CDR3 discussed herein. Further, in some
embodiments, an anti-CSF1R antibody comprises at least one mutated
CDR based on a CDR discussed herein, wherein the mutated CDR
comprises 1, 2, 3, or 4 amino acid substitutions relative to the
CDR discussed herein. In some embodiments, one or more of the amino
acid substitutions are conservative amino acid substitutions. One
skilled in the art can select one or more suitable conservative
amino acid substitutions for a particular CDR sequence, wherein the
suitable conservative amino acid substitutions are not predicted to
significantly alter the binding properties of the antibody
comprising the mutated CDR.
[0152] Exemplary anti-CSF1R antibodies also include antibodies that
compete for binding to CSF1R with an antibody described herein.
Thus, in some embodiments, an anti-CSF1R antibody is provided that
competes for binding to CSF1R with an antibody selected from Fabs
0301, 0302, and 0311, and bivalent (i.e., having two heavy chains
and two light chains) antibody versions of those Fabs.
[0153] Exemplary Antibody Constant Regions
[0154] In some embodiments, an antibody described herein comprises
one or more human constant regions. In some embodiments, the human
heavy chain constant region is of an isotype selected from IgA,
IgG, and IgD. In some embodiments, the human light chain constant
region is of an isotype selected from .kappa. and .lamda.. In some
embodiments, an antibody described herein comprises a human IgG
constant region. In some embodiments, an antibody described herein
comprises a human IgG4 heavy chain constant region. In some such
embodiments, an antibody described herein comprises an S241P
mutation in the human IgG4 constant region. In some embodiments, an
antibody described herein comprises a human IgG4 constant region
and a human .kappa. light chain.
[0155] As noted above, whether or not effector function is
desirable may depend on the particular method of treatment intended
for an antibody. Thus, in some embodiments, when effector function
is desirable, an anti-CSF1R antibody comprising a human IgG1 heavy
chain constant region or a human IgG3 heavy chain constant region
is selected. In some embodiments, when effector function is not
desirable, an anti-CSF1R antibody comprising a human IgG4 or IgG2
heavy chain constant region is selected.
[0156] Exemplary Anti-CSF1R Heavy Chain Variable Regions
[0157] In some embodiments, anti-CSF1R antibody heavy chain
variable regions are provided. In some embodiments, an anti-CSF1R
antibody heavy chain variable region is a mouse variable region, a
human variable region, or a humanized variable region.
[0158] An anti-CSF1R antibody heavy chain variable region comprises
a heavy chain CDR1, FR2, CDR2, FR3, and CDR3. In some embodiments,
an anti-CSF1R antibody heavy chain variable region further
comprises a heavy chain FR1 and/or FR4. Nonlimiting exemplary heavy
chain variable regions include, but are not limited to, heavy chain
variable regions having an amino acid sequence selected from SEQ ID
NOs: 9, 11, 13, and 39 to 45.
[0159] In some embodiments, an anti-CSF1R antibody heavy chain
variable region comprises a CDR1 comprising a sequence selected
from SEQ ID NOs: 15, 21, and 27.
[0160] In some embodiments, an anti-CSF1R antibody heavy chain
variable region comprises a CDR2 comprising a sequence selected
from SEQ ID NOs: 16, 22, and 28.
[0161] In some embodiments, an anti-CSF1R antibody heavy chain
variable region comprises a CDR3 comprising a sequence selected
from SEQ ID NOs: 17, 23, and 29.
[0162] Nonlimiting exemplary heavy chain variable regions include,
but are not limited to, heavy chain variable regions comprising
sets of CDR1, CDR2, and CDR3 selected from: SEQ ID NOs: 15, 16, and
17; SEQ ID NOs: 21, 22, and 23; and SEQ ID NOs: 27, 28, and 29.
[0163] In some embodiments, an anti-CSF1R antibody heavy chain
comprises a variable region sequence that is 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% identical to
a sequence selected from SEQ ID NOs: 9, 11, 13, and 39 to 45,
wherein the heavy chain, together with a light chain, is capable of
forming an antibody that binds CSF1R.
[0164] In some embodiments, an anti-CSF1R antibody heavy chain
comprises at least one of the CDRs discussed herein. That is, in
some embodiments, an anti-CSF1R antibody heavy chain comprises at
least one CDR selected from a heavy chain CDR1 discussed herein, a
heavy chain CDR2 discussed herein, and a heavy chain CDR3 discussed
herein. Further, in some embodiments, an anti-CSF1R antibody heavy
chain comprises at least one mutated CDR based on a CDR discussed
herein, wherein the mutated CDR comprises 1, 2, 3, or 4 amino acid
substitutions relative to the CDR discussed herein. In some
embodiments, one or more of the amino acid substitutions are
conservative amino acid substitutions. One skilled in the art can
select one or more suitable conservative amino acid substitutions
for a particular CDR sequence, wherein the suitable conservative
amino acid substitutions are not predicted to significantly alter
the binding properties of the heavy chain comprising the mutated
CDR.
[0165] In some embodiments, a heavy chain comprises a heavy chain
constant region. In some embodiments, a heavy chain comprises a
human heavy chain constant region. In some embodiments, the human
heavy chain constant region is of an isotype selected from IgA,
IgG, and IgD. In some embodiments, the human heavy chain constant
region is an IgG constant region. In some embodiments, a heavy
chain comprises a human igG4 heavy chain constant region. In some
such embodiments, the human IgG4 heavy chain constant region
comprises an S241P mutation.
[0166] In some embodiments, when effector function is desirable, a
heavy chain comprises a human IgG1 or IgG3 heavy chain constant
region. In some embodiments, when effector function is less
desirable, a heavy chain comprises a human IgG4 or IgG2 heavy chain
constant region.
[0167] Exemplary Anti-CSF1R Light Chain Variable Regions
[0168] In some embodiments, anti-CSF1R antibody light chain
variable regions are provided. In some embodiments, an anti-CSF1R
antibody light chain variable region is a mouse variable region, a
human variable region, or a humanized variable region.
[0169] An anti-CSF1R antibody light chain variable region comprises
a light chain CDR1, FR2, CDR2, FR3, and CDR3. In some embodiments,
an anti-CSF1R antibody light chain variable region further
comprises a light chain FR1 and/or FR4. Nonlimiting exemplary light
chain variable regions include light chain variable regions having
an amino acid sequence selected from SEQ ID NOs: 10, 12, 14, and 46
to 52.
[0170] In some embodiments, an anti-CSF1R antibody light chain
variable region comprises a CDR1 comprising a sequence selected
from SEQ ID NOs: 18, 24 and 30.
[0171] In some embodiments, an anti-CSF1R antibody light chain
variable region comprises a CDR2 comprising a sequence selected
from SEQ ID NOs: 19, 25, and 31.
[0172] In some embodiments, an anti-CSF1R antibody light chain
variable region comprises a CDR3 comprising a sequence selected
from SEQ ID NOs: 20, 26, and 32.
[0173] Nonlimiting exemplary light chain variable regions include,
but are not limited to, light chain variable regions comprising
sets of CDR1, CDR2, and CDR3 selected from: SEQ ID NOs: 18, 19, and
20; SEQ ID NOs: 24, 25, and 26; and SEQ ID NOs: 30, 31, and 32.
[0174] In some embodiments, an anti-CSF1R antibody light chain
comprises a variable region sequence that is 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% identical to
a sequence selected from SEQ ID NOs: 10, 12, 14, and 46 to 52,
wherein the light chain, together with a heavy chain, is capable of
forming an antibody that binds CSF1R.
[0175] In some embodiments, an anti-CSF1R antibody light chain
comprises at least one of the CDRs discussed herein. That is, in
some embodiments, an anti-CSF1R antibody light chain comprises at
least one CDR selected from a light chain CDR1 discussed herein, a
light chain CDR2 discussed herein, and a light chain CDR3 discussed
herein. Further, in some embodiments, an anti-CSF1R antibody light
chain comprises at least one mutated CDR based on a CDR discussed
herein, wherein the mutated CDR comprises 1, 2, 3, or 4 amino acid
substitutions relative to the CDR discussed herein. In some
embodiments, one or more of the amino acid substitutions are
conservative amino acid substitutions. One skilled in the art can
select one or more suitable conservative amino acid substitutions
for a particular CDR sequence, wherein the suitable conservative
amino acid substitutions are not predicted to significantly alter
the binding properties of the light chain comprising the mutated
CDR.
[0176] In some embodiments, a light chain comprises a human light
chain constant region. In some embodiments, a human light chain
constant region is selected from a human .kappa. and a human
.lamda., light chain constant region.
[0177] Exemplary Additional CSF1R Binding Molecules
[0178] In some embodiments, additional molecules that bind CSF1R
are provided. Such molecules include, but are not limited to,
non-canonical scaffolds, such as anti-calins, adnectins, ankyrin
repeats, etc. See, e.g., Hosse et al., Prot. Sci. 15:14 (2006);
Fiedler, M. and Skerra, A., "Non-Antibody Scaffolds," pp. 467-499
in Handbook of Therapeutic Antibodies, Dubel, S., ed., Wiley-VCH,
Weinheim, Germany, 2007.
Exemplary Properties of Anti-CSF1R Antibodies
[0179] In some embodiments, an antibody having a structure
described above binds to the CSF1R with a binding affinity
(K.sub.D) of less than 1 nM, blocks binding of CSF1 and/or IL-34 to
CSF1R, and inhibits CSF1R phosphorylation induced by CSF1 and/or
IL-34.
[0180] In some embodiments, an anti-CSF1R antibody binds to the
extracellular domain of CSF1R (CSF1R-ECD). In some embodiments, an
anti-CSF1R antibody has a binding affinity (K.sub.D) for CSF1R of
less than 1 nM, less than 0.5 nM, less than 0.1 nM, or less than
0.05 nM. In some embodiments, an anti-CSF1R antibody has a K.sub.D
of between 0.01 and 1 nM, between 0.01 and 0.5 nM, between 0.01 and
0.1 nM, between 0.01 and 0.05 nM, or between 0.02 and 0.05 nM.
[0181] In some embodiments, an anti-CSF1R antibody blocks ligand
binding to CSF1R. In some embodiments, an anti-CSF1R antibody
blocks binding of CSF1 to CSF1R. In some embodiments, an anti-CSF1R
antibody blocks binding of IL-34 to CSF1R. In some embodiments, an
anti-CSF1R antibody blocks binding of both CSF1 and IL-34 to CSF1R.
In some embodiments, an antibody that blocks ligand binding binds
to the extracellular domain of CSF1R. An antibody is considered to
"block ligand binding to CSF1R" when it reduces the amount of
detectable binding of a ligand to CSF1R by at least 50%, using the
assay described in Example 7. In some embodiments, an antibody
reduces the amount of detectable binding of a ligand to CSF1R by at
least 60%, at least 70%, at least 80%, or at least 90%, using the
assay described in Example 7. In some such embodiments, the
antibody is said to block ligand binding by at least 50%, at least
60%, at least 70%, etc.
[0182] In some embodiments, an anti-CSF1R antibody inhibits
ligand-induced CSF1R phosphorylation. In some embodiments, an
anti-CSF1R antibody inhibits CSF1-induced CSF1R phosphorylation. In
some embodiments, an anti-CSF1R antibody inhibits IL-34-induced
CSF1R phosphorylation. In some embodiments, an anti-CSF1R antibody
inhibits both CSF1-induced and IL-34-induced CSF1R phosphorylation.
An antibody is considered to "inhibit ligand-induced CSF1R
phosphorylation" when it reduces the amount of detectable
ligand-induced CSF1R phosphorylation by at least 50%, using the
assay described in Example 6. In some embodiments, an antibody
reduces the amount of detectable ligand-induced CSF1R
phosphorylation by at least 60%, at least 70%, at least 80%, or at
least 90%, using the assay described in Example 6. In some such
embodiments, the antibody is said to inhibit ligand-induced CSF1R
phosphorylation by at least at least 50%, at least 60%, at least
70%, etc.
[0183] In some embodiments, an antibody inhibits monocyte
proliferation and/or survival responses in the presence of CSF1
and/or IL-34. An antibody is considered to "inhibit monocyte
proliferation and/or survival responses" when it reduces the amount
of monocyte proliferation and/or survival responses in the presence
of CSF1 and/or IL-34 by at least 50%, using the assay described in
Example 10. In some embodiments, an antibody reduces the amount of
monocyte proliferation and/or survival responses in the presence of
CSF1 and/or IL-34 by at least 60%, at least 70%, at least 80%, or
at least 90%, using the assay described in Example 10. In some such
embodiments, the antibody is said to inhibit monocyte proliferation
and/or survival responses by at least at least 50%, at least 60%,
at least 70%, etc.
Exemplary Antibody Conjugates
[0184] In some embodiments, an anti-CSF1R antibody is conjugated to
a label and/or a cytotoxic agent. As used herein, a label is a
moiety that facilitates detection of the antibody and/or
facilitates detection of a molecule to which the antibody binds.
Nonlimiting exemplary labels include, but are not limited to,
radioisotopes, fluorescent groups, enzymatic groups,
chemiluminescent groups, biotin, epitope tags, metal-binding tags,
etc. One skilled in the art can select a suitable label according
to the intended application.
[0185] As used herein, a cytotoxic agent is a moiety that reduces
the proliferative capacity of one or more cells. A cell has reduced
proliferative capacity when the cell becomes less able to
proliferate, for example, because the cell undergoes apoptosis or
otherwise dies, the cell fails to proceed through the cell cycle
and/or fails to divide, the cell differentiates, etc. Nonlimiting
exemplary cytotoxic agents include, but are not limited to,
radioisotopes, toxins, and chemotherapeutic agents. One skilled in
the art can select a suitable cytotoxic according to the intended
application.
[0186] In some embodiments, a label and/or a cytotoxic agent is
conjugated to an antibody using chemical methods in vitro.
Nonlimiting exemplary chemical methods of conjugation are known in
the art, and include services, methods and/or reagents commercially
available from, e.g., Thermo Scientific Life Science Research
Produces (formerly Pierce; Rockford, Ill.), Prozyme (Hayward,
Calif.), SACRI Antibody Services (Calgary, Canada), AbD Serotec
(Raleigh, N.C.), etc. In some embodiments, when a label and/or
cytotoxic agent is a polypeptide, the label and/or cytotoxic agent
can be expressed from the same expression vector with at least one
antibody chain to produce a polypeptide comprising the label and/or
cytotoxic agent fused to an antibody chain. One skilled in the art
can select a suitable method for conjugating a label and/or
cytotoxic agent to an antibody according to the intended
application.
Exemplary Leader Sequences
[0187] In order for some secreted proteins to express and secrete
in large quantities, a leader sequence from a heterologous protein
may be desirable. In some embodiments, a leader sequence is
selected from SEQ ID NOs: 3 and 4, which are light chain and heavy
chain leader sequences, respectively. In some embodiments,
employing heterologous leader sequences may be advantageous in that
a resulting mature polypeptide may remain unaltered as the leader
sequence is removed in the ER during the secretion process. The
addition of a heterologous leader sequence may be required to
express and secrete some proteins.
[0188] Certain exemplary leader sequence sequences are described,
e.g., in the online Leader sequence Database maintained by the
Department of Biochemistry, National University of Singapore. See
Choo et al., BMC Bioinformatics, 6: 249 (2005); and PCT Publication
No. WO 2006/081430.
Nucleic Acid Molecules Encoding Anti-CSF1R Antibodies
[0189] Nucleic acid molecules comprising polynucleotides that
encode one or more chains of anti-CSF1R antibodies are provided. In
some embodiments, a nucleic acid molecule comprises a
polynucleotide that encodes a heavy chain or a light chain of an
anti-CSF1R antibody. In some embodiments, a nucleic acid molecule
comprises both a polynucleotide that encodes a heavy chain and a
polynucleotide that encodes a light chain, of an anti-CSF1R
antibody. In some embodiments, a first nucleic acid molecule
comprises a first polynucleotide that encodes a heavy chain and a
second nucleic acid molecule comprises a second polynucleotide that
encodes a light chain.
[0190] In some such embodiments, the heavy chain and the light
chain are expressed from one nucleic acid molecule, or from two
separate nucleic acid molecules, as two separate polypeptides. In
some embodiments, such as when an antibody is an scFv, a single
polynucleotide encodes a single polypeptide comprising both a heavy
chain and a light chain linked together.
[0191] In some embodiments, a polynucleotide encoding a heavy chain
or light chain of an anti-CSF1R antibody comprises a nucleotide
sequence that encodes a leader sequence, which, when translated, is
located at the N terminus of the heavy chain or light chain. As
discussed above, the leader sequence may be the native heavy or
light chain leader sequence, or may be another heterologous leader
sequence.
[0192] Nucleic acid molecules may be constructed using recombinant
DNA techniques conventional in the art. In some embodiments, a
nucleic acid molecule is an expression vector that is suitable for
expression in a selected host cell.
Anti-CSF1R Antibody Expression and Production
[0193] Vectors
[0194] Vectors comprising polynucleotides that encode anti-CSF1R
heavy chains and/or anti-CSF1R light chains are provided. Vectors
comprising polynucleotides that encode anti-CSF1R heavy chains
and/or anti-CSF1R light chains are also provided. Such vectors
include, but are not limited to, DNA vectors, phage vectors, viral
vectors, retroviral vectors, etc. In some embodiments, a vector
comprises a first polynucleotide sequence encoding a heavy chain
and a second polynucleotide sequence encoding a light chain. In
some embodiments, the heavy chain and light chain are expressed
from the vector as two separate polypeptides. In some embodiments,
the heavy chain and light chain are expressed as part of a single
polypeptide, such as, for example, when the antibody is an
scFv.
[0195] In some embodiments, a first vector comprises a
polynucleotide that encodes a heavy chain and a second vector
comprises a polynucleotide that encodes a light chain. In some
embodiments, the first vector and second vector are transfected
into host cells in similar amounts (such as similar molar amounts
or similar mass amounts). In some embodiments, a mole- or
mass-ratio of between 5:1 and 1:5 of the first vector and the
second vector is transfected into host cells. In some embodiments,
a mass ratio of between 1:1 and 1:5 for the vector encoding the
heavy chain and the vector encoding the light chain is used. In
some embodiments, a mass ratio of 1:2 for the vector encoding the
heavy chain and the vector encoding the light chain is used.
[0196] In some embodiments, a vector is selected that is optimized
for expression of polypeptides in CHO or CHO-derived cells, or in
NSO cells. Exemplary such vectors are described, e.g., in Running
Deer et al., Biotechnol. Prog. 20:880-889 (2004).
[0197] In some embodiments, a vector is chosen for in vivo
expression of anti-CSF1R heavy chains and/or anti-CSF1R light
chains in animals, including humans. In some such embodiments,
expression of the polypeptide is under the control of a promoter
that functions in a tissue-specific manner. For example,
liver-specific promoters are described, e.g., in PCT Publication
No. WO 2006/076288.
[0198] Host Cells
[0199] In various embodiments, anti-CSF1R heavy chains and/or
anti-CSF1R light chains may be expressed in prokaryotic cells, such
as bacterial cells; or in eukaryotic cells, such as fungal cells
(such as yeast), plant cells, insect cells, and mammalian cells.
Such expression may be carried out, for example, according to
procedures known in the art. Exemplary eukaryotic cells that may be
used to express polypeptides include, but are not limited to, COS
cells, including COS 7 cells; 293 cells, including 293-6E cells;
CHO cells, including CHO-S and DG44 cells; PER.C6.RTM. cells
(Crucell); and NSO cells. In some embodiments, anti-CSF1R heavy
chains and/or anti-CSF1R light chains may be expressed in yeast.
See, e.g., U.S. Publication No. US 2006/0270045 A1. In some
embodiments, a particular eukaryotic host cell is selected based on
its ability to make desired post-translational modifications to the
anti-CSF1R heavy chains and/or anti-CSF1R light chains. For
example, in some embodiments, CHO cells produce polypeptides that
have a higher level of sialylation than the same polypeptide
produced in 293 cells.
[0200] Introduction of one or more nucleic acids into a desired
host cell may be accomplished by any method, including but not
limited to, calcium phosphate transfection, DEAE-dextran mediated
transfection, cationic lipid-mediated transfection,
electroporation, transduction, infection, etc. Nonlimiting
exemplary methods are described, e.g., in Sambrook et al.,
Molecular Cloning, A Laboratory Manual, 3.sup.rd ed. Cold Spring
Harbor Laboratory Press (2001). Nucleic acids may be transiently or
stably transfected in the desired host cells, according to any
suitable method.
[0201] In some embodiments, one or more polypeptides may be
produced in vivo in an animal that has been engineered or
transfected with one or more nucleic acid molecules encoding the
polypeptides, according to any suitable method.
[0202] Purification of Anti-CSF1R Antibodies
[0203] Anti-CSF1R antibodies may be purified by any suitable
method. Such methods include, but are not limited to, the use of
affinity matrices or hydrophobic interaction chromatography.
Suitable affinity ligands include the CSF1R ECD and ligands that
bind antibody constant regions. For example, a Protein A, Protein
G, Protein A/G, or an antibody affinity column may be used to bind
the constant region and to purify an anti-CSF1R antibody.
Hydrophobic interactive chromatography, for example, a butyl or
phenyl column, may also suitable for purifying some polypeptides.
Many methods of purifying polypeptides are known in the art.
[0204] Cell-Free Production of Anti-CSF1R Antibodies
[0205] In some embodiments, an anti-CSF1R antibody is produced in a
cell-free system. Nonlimiting exemplary cell-free systems are
described, e.g., in Sitaraman et al., Methods Mol. Biol. 498:
229-44 (2009); Spirin, Trends Biotechnol. 22: 538-45 (2004); Endo
et al., Biotechnol. Adv. 21: 695-713 (2003).
Therapeutic Compositions and Methods
[0206] Methods of Treating Diseases Using Anti-CSF1R Antibodies
[0207] Provided herein are methods of treating systemic lupus
erythematosus (SLE), rheumatoid arthritis, and multiple sclerosis
with an antibody that binds CSF1R and blocks CSF1 and IL-34 ligand
binding.
[0208] In some embodiments, a method of treating SLE is provided.
In some such embodiments, the method comprises administering an
antibody that binds CSF1R and blocks CSF1 and IL-34 ligand binding,
such as an antibody selected from huAb1 to huAb16, to a subject
with SLE. Treating SLE, in some embodiments, means reducing the
incidence and/or severity of skin lesions associated with SLE,
and/or reducing a kidney condition selected from
glomerulonephritis, interstitial nephritis, and perivascular
infiltrates. In some embodiments, a method of treating lupus
nephritis is provided, wherein the method comprises administering
an antibody that binds CSF1R and blocks CSF1 and IL-34 ligand
binding, such as an antibody selected from huAb1 to huAb16, to a
subject with lupus nephritis. In some embodiments, a method of
treating a class of lupus nephritis selected from class I, class
II, class III, class IV, class V, and class VI lupus nephritis is
provided, wherein the method comprises administering an antibody
that binds CSF1R and blocks CSF1 and IL-34 ligand binding, such as
an antibody selected from huAb1 to huAb16, to a subject with lupus
nephritis. Class III and class IV lupus nephritis have
significantly greater CD16+ monocyte counts than other classes of
lupus nephritis. See Yoshimoto et al., Am. J. Kidney Dis. 50: 47-58
(2007). Accordingly, in some embodiments, a method of treating
class III or class IV lupus nephritis is provided, wherein the
method comprises administering an antibody that binds CSF1R and
blocks CSF1 and IL-34 ligand binding, such as an antibody selected
from huAb1 to huAb16, to a subject with lupus nephritis. In some
such embodiments, the method comprises administering huAb1 to the
subject.
[0209] In some embodiments, a method of treating skin lesions
associated with SLE is provided, wherein the method comprises
administering an antibody that binds CSF1R, such as an antibody
selected from huAb1 to huAb16, to a subject with SLE. In some
embodiments, a method of treating skin lesions associated with SLE
comprises administering huAb1 to a subject with SLE. In some
embodiments, a method of reducing the number, severity, and/or rate
of formation of skin lesions associated with SLE is provided,
wherein the method comprises administering an antibody that binds
CSF1R, such as an antibody selected from huAb1 to huAb16, to a
subject with SLE. In some embodiments, a method of reducing the
number, severity, and/or rate of formation of skin lesions
associated with SLE comprises administering huAb1 to a subject with
SLE. The number, severity, and/or rate of formation of skin lesions
may be determined, in some embodiments, by visual inspection. Such
visual inspection may be carried out, for example, by a clinician
or by the subject (e.g., self-reported visual inspection).
[0210] In some embodiments, a method of treating a kidney condition
associated with SLE is provided. In some such embodiments, the
method comprises administering an antibody that binds CSF1R and
blocks CSF1 and IL-34 ligand binding, such as an antibody selected
from huAb1 to huAb16, to a subject with a kidney condition
associated with SLE. In some embodiments, antibody huAb1 is
administered to the subject. In some embodiments, the kidney
condition is selected from glomerulonephritis, interstitial
nephritis, and perivascular infiltrates. In some embodiments, a
method of treating lupus nephritis is provided, wherein the method
comprises administering an antibody that binds CSF1R and blocks
CSF1 and IL-34 ligand binding, such as an antibody selected from
huAb1 to huAb16, to a subject with lupus nephritis. In some
embodiments, a method of treating lupus nephritis comprises
administering antibody huAb1 to a subject with lupus nephritis. The
extent of kidney damage in lupus nephritis may be determined, in
some embodiments, using ultrasonography, scintigraphy, pyelography,
or needle biopsy. The extent of kidney damage in lupus nephritis
may also be determined, in some embodiments, by measuring kidney
function, such as, for example, by measuring proteinuria and/or
glomerular filtration rate.
[0211] In some embodiments, a method of reducing proteinuria
associated with lupus nephritis is provided, comprising
administering an antibody that binds CSF1R and blocks CSF1 and
IL-34 ligand binding, such as an antibody selected from huAb1 to
huAb16, to a subject exhibiting proteinuria. Proteinuria may be
detected by any method in the art, including, but not limited to,
dipstick testing, protein/creatinine ratio, serum
albumin/creatinine ratio, and protein electrophoresis. Levels of
protein in urine may be indicated, in some embodiments, using the
following scale: trace (5-20 mg/dl), 1+ (30 mg/dl), 2+ (100 mg/dl),
3+ (300 mg/dl), and 4+ (>2000 mg/dl). A protein/creatinine ratio
of <200 mg/g is considered normal. A serum albumin/creatinine
ratio of <17 mg/g and <25 mg/g is considered normal for men
and women, respectively. Proteinuria is defined by the National
Kidney Foundation Kidney Disease Outcome Quality Initiative as the
presence of >300 mg/d total protein or albumin for 24-hour urine
excretions; as >30 mg/dL total protein for spot urine dipstick
measurements (i.e., 1+ or greater on the scale described above); as
>200 mg/g total protein-to-creatinine ratio; as >300 mg/g
albumin-to-creatinine ratio for spot urine measurements (or >250
mg/g albumin-to-creatinine ratio for spot urine measurements from
men; and >355 mg/g albumin-to-creatinine ratio for spot urine
measurements from women).
[0212] In some embodiments, administration of an antibody that
binds CSF1R and blocks CSF1 and IL-34 ligand binding to a subject
reduces proteinuria by at least one number or at least two numbers
on the "trace, 1+, 2+, 3+, 4+" scale. In other words, in some
embodiments, administration of an antibody that binds CSF1R and
blocks CSF1 and IL-34 ligand binding to a subject with 3+
proteinuria reduces the proteinuria to a 2+ level or lower, or a 1+
level or lower. In some embodiments, administration of an antibody
that binds CSF1R and blocks CSF1 and IL-34 ligand binding to a
subject reduces the protein/creatinine ratio and/or
albumin/creatinine ratio by at least 50 mg/g, at least 100 mg/g, at
least 150 mg/g, at least 200 mg/g, at least 250 mg/g, or at least
300 mg/g. In some embodiments, administration of an antibody that
binds CSF1R and blocks CSF1 and IL-34 ligand binding to a subject
reduces the protein/creatinine ratio and/or albumin/creatinine
ratio to below 250 mg/g, below 200 mg/g, below 150 mg/g, or below
100 mg/g. In some embodiments, administration of an antibody that
binds CSF1R and blocks CSF1 and IL-34 ligand binding to a subject
reduces the protein/creatinine ratio and/or albumin/creatinine
ratio to within 2-fold, within 3-fold, within 4-fold, within 5-fold
of a normal ratio. In some embodiments, the reduction in
proteinuria is determined in the first week, in the first two
weeks, in the first month, in the first two months, in the first
three months, in the first 4 months, or in the first six months
after beginning treatment. In some embodiments, a subject is
monitored for continued suppression of proteinuria while being
treated with an antibody that binds CSF1R and blocks CSF1 and IL-34
ligand binding.
[0213] In some embodiments, a method of improving glomerular
filtration rate in a subject with lupus nephritis is provided,
comprising administering an antibody that binds CSF1R and blocks
CSF1 and IL-34 ligand binding, such as an antibody selected from
huAb1 to huAb16 to a subject with lupus nephritis. Glomerular
filtration rate may be determined by any method in the art,
including, but not limited to, serum or urine levels of insulin or
sinistrin (in some embodiments, insulin or sinistrin is injected
prior to analysis), creatinine clearance estimated from creatinine
concentrations in 24 urine collection and serum, and serum cystatin
C levels. Normal glomerular filtration rates in adults are in the
range of 90-130 ml/min/1.73 m.sup.2. Glomerular filtration rates
below 90 ml/min/1.73 m.sup.2, below 80 ml/min/1.73 m.sup.2, below
70 ml/min/1.73 m.sup.2, or below 60 ml/min/1.73 m.sup.2 are, in
some embodiments, indicative of chronic kidney disease, while rates
below 15 ml/min/1.73 m.sup.2 are indicative of kidney failure. In
some embodiments, a method of improving glomerular filtration rate
in a subject with lupus nephritis is provided, comprising
administering an antibody that binds CSF1R and blocks CSF1 and
IL-34 ligand binding, such as an antibody selected from huAb1 to
huAb16, to a subject with lupus nephritis. In some embodiments,
following administration of an antibody that binds CSF1R and blocks
CSF1 and IL-34 ligand binding, a subject's glomerular filtration
rate improves by at least 10 ml/min/1.73 m.sup.2, at least 15
ml/min/1.73 m.sup.2, at least 20 ml/min/1.73 m.sup.2, at least 25
ml/min/1.73 m.sup.2, at least 30 ml/min/1.73 m.sup.2, at least 40
ml/min/1.73 m.sup.2, or at least 50 ml/min/1.73 m.sup.2. In some
embodiments, following administration of an antibody that binds
CSF1R and blocks CSF1 and IL-34 ligand binding, a subject's
glomerular filtration rate is at least 60 ml/min/1.73 m.sup.2, at
least 70 ml/min/1.73 m.sup.2, at least 80 ml/min/1.73 m.sup.2, or
at least 90 ml/min/1.73 m.sup.2. In some embodiments, the reduction
in glomerular filtration rate is determined in the first week, in
the first two weeks, in the first month, in the first two months,
in the first three months, in the first 4 months, or in the first
six months after beginning treatment. In some embodiments, a
subject is monitored for continued improvement of glomerular
filtration rate while being treated with an antibody that binds
CSF1R and blocks CSF1 and IL-34 ligand binding.
[0214] In some embodiments, a method of treating lupus nephritis is
provided, comprising administering an antibody that binds CSF1R and
blocks CSF1 and IL-34 ligand binding, such as an antibody selected
from huAb1 to huAb16 to a subject with lupus nephritis, wherein
administration of the antibody slows or prevents the progression of
a kidney condition associated with lupus nephritis. In some such
embodiments, proteinuria and/or the glomerular filtration rate in
the subject does not worsen (i.e., proteinuria does not increase
and/or the glomerular filtration rate does not decrease) following
administration of the antibody and/or during a particular time
interval during which the subject is undergoing treatment with the
antibody. The treatment may be a single dose or multiple doses.
[0215] In some embodiments, methods of treating rheumatoid
arthritis are provided, wherein the method comprises administering
an antibody that binds CSF1R and blocks CSF1 and IL-34 ligand
binding, such as an antibody selected from huAb1 to huAb16, to a
subject with rheumatoid arthritis. In some embodiments, methods of
treating rheumatoid arthritis are provided, wherein the method
comprises administering antibody huAb1 to a subject with rheumatoid
arthritis. Treating rheumatoid arthritis, in some embodiments,
means reducing clinical disease scores, which may be measured, for
example, by measuring erythema and swelling in joints affected by
rheumatoid arthritis. Treating rheumatoid arthritis, in some
embodiments, means reducing inflammation, reducing pannus
formation, reducing cartilage damage, reducing bone resorption,
reducing macrophage numbers in the joints, reducing autoantibody
formation, and/or reducing bone loss.
[0216] In some embodiments, a method of reducing inflammation
associated with rheumatoid arthritis is provided, wherein the
method comprises administering an antibody that binds CSF1R and
blocks CSF1 and IL-34 ligand binding, such as an antibody selected
from huAb1 to huAb16, to a subject with rheumatoid arthritis. In
some embodiments, a method of reducing inflammation associated with
rheumatoid arthritis is provided, wherein the method comprises
administering antibody huAb1 to a subject with rheumatoid
arthritis. Reducing inflammation, in some embodiments, comprises
reducing erythrocyte sedimentation rate and/or reducing the levels
of C-reactive proteins in blood. When inflammation is present in a
subject, the erythrocyte sedimentation rate increases, possibly due
to increased levels of fibrinogen in the blood. The erythrocyte
sedimentation rate may be determined by any method in the art,
including, but not limited to, calculating the rate by measuring
the change in height of anticoagulated erythrocytes in one hour in
a Westergren tube. See also Procedures for the Erythrocyte
Sedimentation Rate Test; Approved Standard--Fifth Edition, CLSI
document H02-A5. Wayne, Pa.: Clinical and Laboratory Standards
institute; 2011. Levels of C-reactive protein in blood may be
determined by any methods in the art, including but not limited to
using the RAPITEX.RTM. CRP test kit (Siemens).
[0217] Reducing inflammation, in some embodiments, comprises
reducing peripheral edema, which is tissue swelling due to the
buildup of fluids. Peripheral edema may occur, in some instances,
in the ankles, feet, legs, and/or calves of a subject with
rheumatoid arthritis. Reducing inflammation, in some embodiments,
comprises reducing infiltration of inflammatory cells in the
synovium of one or more affected joints. Synovial fluid may be
collected, in some embodiments, by athrocentesis.
[0218] In some embodiments, a method of reducing pannus formation
associated with rheumatoid arthritis is provided, wherein the
method comprises administering an antibody that binds CSF1R and
blocks CSF1 and IL-34 ligand binding, such as an antibody selected
from huAb1 to huAb16, to a subject with rheumatoid arthritis. In
some embodiments, a method of reducing pannus formation associated
with rheumatoid arthritis is provided, wherein the method comprises
administering antibody huAb1 to a subject with rheumatoid
arthritis. Reducing pannus formation, in some embodiments,
comprises reducing infiltration of pannus into cartilage and/or
subchondrial bone, and/or reducing hard tissue destruction
resulting from pannus infiltration. Pannus formation can be
measured by any method in the art, including, but not limited to,
imaging one or more affected joints. Nonlimiting exemplary imaging
techniques for detecting pannus formation include magnetic
resonance imaging (MM), computed tomography (CT) scan, arthroscopy,
ultrasonography, duplex ultrasonography, and power doppler
imaging.
[0219] In some embodiments, a method of slowing the progression of
pannus formation associated with rheumatoid arthritis is provided,
wherein the method comprises administering an antibody that binds
CSF1R and blocks CSF1 and IL-34 ligand binding, such as an antibody
selected from huAb1 to huAb16, to a subject with rheumatoid
arthritis. In some embodiments, the antibody is huAb1. In some
embodiments, the formation of pannus is slower following
administration of the antibody and/or during a particular time
interval during which the subject is undergoing treatment with the
antibody. The treatment may be a single dose or multiple doses.
[0220] In some embodiments, a method of reducing cartilage damage
associated with rheumatoid arthritis is provided, wherein the
method comprises administering an antibody that binds CSF1R and
blocks CSF1 and IL-34 ligand binding, such as an antibody selected
from huAb1 to huAb16, to a subject with rheumatoid arthritis. In
some embodiments, a method of reducing cartilage damage associated
with rheumatoid arthritis is provided, wherein the method comprises
administering antibody huAb1 to a subject with rheumatoid
arthritis. Reducing cartilage damage, in some embodiments,
comprises reducing chondrocyte loss, reducing collagen disruption,
and/or reducing cartilage loss. Cartilage damage can be measured by
any method in the art, including, but not limited to, imaging one
or more affected joints. Nonlimiting exemplary imaging techniques
for detecting cartilage damage include MM, CT scan, arthroscopy,
and x-ray imaging.
[0221] In some embodiments, a method of slowing the progression of
cartilage damage associated with rheumatoid arthritis is provided,
wherein the method comprises administering an antibody that binds
CSF1R and blocks CSF1 and IL-34 ligand binding, such as an antibody
selected from huAb1 to huAb16, to a subject with rheumatoid
arthritis. In some embodiments, the antibody is huAb1. In some
embodiments, the progression of cartilage damage is slower
following administration of the antibody and/or during a particular
time interval during which the subject is undergoing treatment with
the antibody. The treatment may be a single dose or multiple
doses.
[0222] In some embodiments, a method of reducing bone resorption
associated with rheumatoid arthritis is provided, wherein the
method comprises administering an antibody that binds CSF1R and
blocks CSF1 and IL-34 ligand binding, such as an antibody selected
from huAb1 to huAb16, to a subject with rheumatoid arthritis. In
some embodiments, a method of reducing bone resorption associated
with rheumatoid arthritis is provided, wherein the method comprises
administering antibody huAb1 to a subject with rheumatoid
arthritis. Reducing bone resorption, in some embodiments, comprises
reducing the number of osteoclasts in joints affected by rheumatoid
arthritis.
[0223] In some embodiments, bone resorption may be measured by
determining the level of TRAP5b in plasma from the subject, wherein
an elevated level of TRAP5b indicates elevated bone resorption in
the subject. Thus, in some embodiments, a reduced level of TRAP5b
indicates a reduction in bone resorption. TRAP5b levels may be
determined, in certain instances, before and after treatment with
an antibody that binds CSF1R, and/or may be determined periodically
throughout the course of treatment to monitor the effectiveness of
the treatment in reducing bone loss. TRAP5b levels may be
determined using any method in the art, including, but not limited
to, ELISA (including FAICEA, or fragments absorbed immunocapture
enzymatic assay; see, e.g., Quidel.RTM. TRAP5b assay, TECOmedical
Group, Sissach, Switzerland).
[0224] In some embodiments, a method of reducing bone loss
associated with rheumatoid arthritis is provided, wherein the
method comprises administering an antibody that binds CSF1R and
blocks CSF1 and IL-34 ligand binding, such as an antibody selected
from huAb1 to huAb16, to a subject with rheumatoid arthritis. In
some embodiments, a method of reducing bone loss associated with
rheumatoid arthritis is provided, wherein the method comprises
administering antibody huAb1 to a subject with rheumatoid
arthritis. Bone loss may be determined using any method in the art,
including, but not limited to, x-ray imaging, MM, CT, bone
densitometry, single and dual photon absorptiometry (SPA, DPA),
single and dual energy x-ray absorptiometry (SXA, DXA),
ultrasonography, scintigraphy, and by measuring levels of serum
markers of bone formation and resorption. Nonlimiting exemplary
serum markers of bone formation and bone resorption are shown in
Table 2.
TABLE-US-00002 TABLE 2 Serum markers of bone formation and
resorption Formation Markers Resorption Markers Serum osteocalcin
(OC) Serum and urinary hydroxyproline (Hyp) Serum total alkaline
phosphatase (ALP) Urinary total pyridinoline (Pyr) Serum bone
specific alkaline Urinary total phosphatase (BSAP, BALP, or B-ALP)
deoxypyridinoline (dPyr) Serum procollagen I carboxyterminal
Urinary free pyridinoline (f-Pyr, also propeptide (PICP) known as
Pyrilinks .RTM. (Metra Biosystems)) Serum procollagen type I
N-terminal Urinary free deoxypyridinoline propeptide (PINP)
(f-dPyr, also known as Pyrilinks-D .RTM.) Bone sialoprotein Serum
and urinary collagen type I cross-linked N-telopeptide (NTx, also
referred to as Osteomark) Serum and urinary collagen type I
cross-linked C-terminal telopeptide (CTx, also referred to as
CrossLaps .RTM.) Serum carboxyterminal telopeptide of type I
collagen (ITCP) Tartrate-resistant acid phosphatase (TRAP or
RACP)
[0225] In some embodiments, a method of slowing the progression of
bone loss associated with rheumatoid arthritis is provided, wherein
the method comprises administering an antibody that binds CSF1R and
blocks CSF1 and IL-34 ligand binding, such as an antibody selected
from huAb1 to huAb16, to a subject with rheumatoid arthritis. In
some embodiments, the antibody is huAb1. In some embodiments, bone
loss is slower following administration of the antibody and/or
during a particular time interval during which the subject is
undergoing treatment with the antibody. The treatment may be a
single dose or multiple doses.
[0226] In some embodiments, a method of reducing the number of
monocyte lineage cells, such as macrophages and/or CD16+ monocytes,
in joints affected by rheumatoid arthritis is provided, wherein the
method comprises administering an antibody that binds CSF1R and
blocks CSF1 and IL-34 ligand binding, such as an antibody selected
from huAb1 to huAb16, to a subject with rheumatoid arthritis. In
some embodiments, a method of reducing the number of monocyte
lineage cells, such as macrophages and/or CD16+ monocytes, in
joints affected by rheumatoid arthritis is provided, wherein the
method comprises administering antibody huAb1 to a subject with
rheumatoid arthritis. In some embodiments, a method of reducing the
number of monocyte lineage cells, such as macrophages and/or CD16+
monocytes, in synovial fluid of joints affected by rheumatoid
arthritis is provided, wherein the method comprises administering
an antibody that binds CSF1R and blocks CSF1 and IL-34 ligand
binding, such as an antibody selected from huAb1 to huAb16, to a
subject with rheumatoid arthritis. In some embodiments, a method of
reducing the number of monocyte lineage cells, such as macrophages
and/or CD16+ monocytes, in synovial fluid of joints affected by
rheumatoid arthritis is provided, wherein the method comprises
administering antibody huAb1 to a subject with rheumatoid
arthritis.
[0227] In some embodiments, a method of reducing autoantibody
levels in a subject with rheumatoid arthritis is provided, wherein
the method comprises administering an antibody that binds CSF1R and
blocks CSF1 and IL-34 ligand binding, such as an antibody selected
from huAb1 to huAb16, to a subject with rheumatoid arthritis. In
some embodiments, a method of reducing autoantibody levels in a
subject with rheumatoid arthritis is provided, wherein the method
comprises administering antibody huAb1 to a subject with rheumatoid
arthritis.
[0228] In some embodiments, a method of reducing autoantibody
levels in a subject with rheumatoid arthritis are provided,
comprising administering an antibody that binds CSF1R and blocks
CSF1 and IL-34 ligand binding, such as an antibody selected from
huAb1 to huAb16, to a subject with rheumatoid arthritis. In some
such embodiments, the method comprises administering huAb1 to a
subject with rheumatoid arthritis. The levels of autoantibodies may
be determined by any method in the art. In some embodiments,
autoantibody levels are determined by the level of rheumatoid
factor (RF) and/or anti-citrullinated protein antibodies (ACPA)
and/or anti-nuclear antibodies (ANA).
[0229] In some embodiments, methods of treating multiple sclerosis
are provided, wherein the method comprises administering an
antibody that binds CSF1R and blocks CSF1 and IL-34 ligand binding,
such as an antibody selected from huAb1 to huAb16, to a subject
with multiple sclerosis. Treating multiple sclerosis, in some
embodiments, means reducing clinical disease scores.
[0230] In some embodiments, methods of treating CD16+ disorders are
provided, wherein the method comprises administering an antibody
that binds CSF1R, wherein the antibody blocks binding of CSF1 to
CSF1R and blocks binding of IL-34 to CSF1R, such as an antibody
selected from huAb1 to huAb16, to a subject with a CD16+ disorder.
In some embodiments, the antibody reduces the number of CD16+
monocytes by at least 30%, and CD16- monocytes are not reduced or
are reduced by less than 20%. In some embodiments, CD16+ monocytes
are reduced by at least 50%. In some embodiments, the CD16+
monocytes are CD16+ peripheral blood monocytes. In some
embodiments, the CD16+ disorder is selected from rheumatoid
arthritis, juvenile idiopathic arthritis, psoriasis, psoriatic
arthritis, ankylosing spondylitis, Crohn's disease, and ulcerative
colitis, lupus erythematosus, inflammatory bowel disease, Sjogren's
syndrome, coronary artery disease, asthma, and sarcoidosis.
[0231] In some embodiments, methods of decreasing CD16+ monocytes
are provided, wherein the methods comprise administering an
antibody that binds CSF1R and blocks CSF1 and IL-34 ligand binding,
such as an antibody selected from huAb1 to huAb16, to a subject
with increased CD16+ monocytes. In some embodiments, the subject
has an autoimmune condition selected from rheumatoid arthritis and
SLE. In some embodiments, administering an antibody that binds
CSF1R and blocks CSF1 and IL-34 ligand binding does not decrease
the number of CD16- monocytes. In some embodiments, CD16+ monocytes
are reduced to a greater extent than CD16- monocytes are reduced
when an antibody that binds CSF1R and blocks CSF1 and IL-34 ligand
binding is administered to the subject. In some embodiments, CD16+
monocytes are reduced by at least 20%, at least 30%, at least 50%,
at least 60%, at least 70%, at least 80%, or at least 90%. In some
embodiments, CD16- monocytes are reduced by less than 30%, less
than 20%, or less than 10%. In some embodiments, the CD16+
monocytes are CD16+ peripheral blood monocytes. In some
embodiments, the CD16- monocytes are CD16- peripheral blood
monocytes.
[0232] Routes of Administration and Carriers
[0233] In various embodiments, anti-CSF1R antibodies may be
administered in vivo by various routes, including, but not limited
to, oral, intra-arterial, parenteral, intranasal, intramuscular,
intracardiac, intraventricular, intratracheal, buccal, rectal,
intraperitoneal, intradermal, topical, transdermal, and
intrathecal, or otherwise by implantation or inhalation. The
subject compositions may be formulated into preparations in solid,
semi-solid, liquid, or gaseous forms; including, but not limited
to, tablets, capsules, powders, granules, ointments, solutions,
suppositories, enemas, injections, inhalants, and aerosols. A
nucleic acid molecule encoding an anti-CSF1R antibody may be coated
onto gold microparticles and delivered intradermally by a particle
bombardment device, or "gene gun," as described in the literature
(see, e.g., Tang et al., Nature 356:152-154 (1992)). The
appropriate formulation and route of administration may be selected
according to the intended application.
[0234] In various embodiments, compositions comprising anti-CSF1R
antibodies are provided in formulations with a wide variety of
pharmaceutically acceptable carriers (see, e.g., Gennaro,
Remington: The Science and Practice of Pharmacy with Facts and
Comparisons: Drugfacts Plus, 20th ed. (2003); Ansel et al.,
Pharmaceutical Dosage Forms and Drug Delivery Systems, 7.sup.th
ed., Lippencott Williams and Wilkins (2004); Kibbe et al., Handbook
of Pharmaceutical Excipients, 3.sup.rd ed., Pharmaceutical Press
(2000)). Various pharmaceutically acceptable carriers, which
include vehicles, adjuvants, and diluents, are available. Moreover,
various pharmaceutically acceptable auxiliary substances, such as
pH adjusting and buffering agents, tonicity adjusting agents,
stabilizers, wetting agents and the like, are also available.
Non-limiting exemplary carriers include saline, buffered saline,
dextrose, water, glycerol, ethanol, and combinations thereof.
[0235] In various embodiments, compositions comprising anti-CSF1R
antibodies may be formulated for injection, including subcutaneous
administration, by dissolving, suspending, or emulsifying them in
an aqueous or nonaqueous solvent, such as vegetable or other 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 various
embodiments, the compositions may be formulated for inhalation, for
example, using pressurized acceptable propellants such as
dichlorodifluoromethane, propane, nitrogen, and the like. The
compositions may also be formulated, in various embodiments, into
sustained release microcapsules, such as with biodegradable or
non-biodegradable polymers. A non-limiting exemplary biodegradable
formulation includes poly lactic acid-glycolic acid polymer. A
non-limiting exemplary non-biodegradable formulation includes a
polyglycerin fatty acid ester. Certain methods of making such
formulations are described, for example, in EP 1 125 584 A1.
[0236] Pharmaceutical packs and kits comprising one or more
containers, each containing one or more doses of an anti-CSF1R
antibody are also provided. In some embodiments, a unit dosage is
provided wherein the unit dosage contains a predetermined amount of
a composition comprising an anti-CSF1R antibody, with or without
one or more additional agents. In some embodiments, such a unit
dosage is supplied in single-use prefilled syringe for injection.
In various embodiments, the composition contained in the unit
dosage may comprise saline, sucrose, or the like; a buffer, such as
phosphate, or the like; and/or be formulated within a stable and
effective pH range. Alternatively, in some embodiments, the
composition may be provided as a lyophilized powder that may be
reconstituted upon addition of an appropriate liquid, for example,
sterile water. In some embodiments, the composition comprises one
or more substances that inhibit protein aggregation, including, but
not limited to, sucrose and arginine. In some embodiments, a
composition of the invention comprises heparin and/or a
proteoglycan.
[0237] Pharmaceutical compositions are administered in an amount
effective for treatment or prophylaxis of the specific indication.
The therapeutically effective amount is typically dependent on the
weight of the subject being treated, his or her physical or health
condition, the extensiveness of the condition to be treated, or the
age of the subject being treated. In general, anti-CSF1R antibodies
may be administered in an amount in the range of about 10 .mu.g/kg
body weight to about 100 mg/kg body weight per dose. In some
embodiments, anti-CSF1R antibodies may be administered in an amount
in the range of about 50 .mu.g/kg body weight to about 5 mg/kg body
weight per dose. In some embodiments, anti-CSF1R antibodies may be
administered in an amount in the range of about 100 .mu.g/kg body
weight to about 10 mg/kg body weight per dose. In some embodiments,
anti-CSF1R antibodies may be administered in an amount in the range
of about 100 .mu.g/kg body weight to about 20 mg/kg body weight per
dose. In some embodiments, anti-CSF1R antibodies may be
administered in an amount in the range of about 0.5 mg/kg body
weight to about 20 mg/kg body weight per dose.
[0238] The anti-CSF1R antibody compositions may be administered as
needed to subjects. Determination of the frequency of
administration may be made by persons skilled in the art, such as
an attending physician based on considerations of the condition
being treated, age of the subject being treated, severity of the
condition being treated, general state of health of the subject
being treated and the like. In some embodiments, an effective dose
of an anti-CSF1R antibody is administered to a subject one or more
times. In various embodiments, an effective dose of an anti-CSF1R
antibody is administered to the subject once a month, less than
once a month, such as, for example, every two months or every three
months. In other embodiments, an effective dose of an anti-CSF1R
antibody is administered more than once a month, such as, for
example, every three weeks, every two weeks or every week. An
effective dose of an anti-CSF1R antibody is administered to the
subject at least once. In some embodiments, the effective dose of
an anti-CSF1R antibody may be administered multiple times,
including for periods of at least a month, at least six months, or
at least a year.
[0239] Combination Therapy
[0240] Anti-CSF1R antibodies may be administered alone or with
other modes of treatment. They may be provided before,
substantially contemporaneous with, or after other modes of
treatment, for example, surgery, chemotherapy, radiation therapy,
or the administration of a biologic, such as another therapeutic
antibody. For treatment of rheumatoid arthritis, anti-CSF1R
antibodies may be administered with other therapeutic agents, for
example, methotrexate, anti-TNF agents such as Remicade
(infliximab), Humira (adalimumab), Simponi (golimumab), and Enbrel
(etanercept); glucocorticoids such as prednisone; leflunomide;
azothioprine; JAK inhibitors such as CP 590690; SYK inhibitors such
as R788; anti-IL-6 antibodies; anti-IL-6R antibodies such as
tocilizumab; anti-CD-20 antibodies such as rituximab; anti-CD19
antibodies; anti-GM-CSF antibodies; anti-GM-CSF-R antibodies; IL-1
receptor antagonists such as anakinra; CTLA-4 antagonists, such as
abatacept; immunosuppressants such as cyclosporine.
[0241] For treatment of systemic lupus erythematosus, anti-CSF1R
antibodies may be administered with other therapeutic agents, for
example, hydroxychloroquine (Plaquenil); corticosteroids, such as
prednisone, methylprednisone, and prednisolone; immunosuppressants,
such as cyclophosphamide (Cytoxan), azathioprine (Imuran, Azasan),
mycophenolate (Cellcept), leflunomide (Arava) and methotrexate
(Trexall), and belimumab (Benlysta).
[0242] For treatment of multiple sclerosis, anti-CSF1R antibodies
may be administered with other therapeutic agents, for example,
interferon alpha; interferon beta; prednisone; anti-alpha4 integrin
antibodies such as Tysabri; anti-CD20 antibodies such as Rituxan;
FTY720 (Fingolimod); and Cladribine (Leustatin).
EXAMPLES
[0243] The examples discussed below are intended to be purely
exemplary of the invention and should not be considered to limit
the invention in any way. The examples are not intended to
represent that the experiments below are all or the only
experiments performed. Efforts have been made to ensure accuracy
with respect to numbers used (for example, amounts, temperature,
etc.) but some experimental errors and deviations should be
accounted for. Unless indicated otherwise, parts are parts by
weight, molecular weight is weight average molecular weight,
temperature is in degrees Centigrade, and pressure is at or near
atmospheric.
Example 1: Humanized Anti-CSF1R Antibodies
[0244] Various humanized anti-CSF1R antibodies were developed
previously. See, e.g., PCT Publication No. WO 2011/140249.
[0245] The sequences for each of the humanized heavy chain variable
regions and humanized light chain variable regions, aligned with
the sequences of the parental chimeric antibody variable regions
and the sequences of the human acceptor variable framework regions
are shown in FIGS. 1 (heavy chains) and 2 (light chains). The
changes in humanized variable region sequences relative to the
human acceptor variable framework region sequences are boxed. Each
of the CDRs for each of the variable regions is shown in a boxed
region, and labeled as "CDR" above the boxed sequences.
[0246] Table 5, below, shows the full sequences for the humanized
heavy chains and humanized light chains of antibodies huAb1 to
huAb16. The name and SEQ ID NOs of the humanized heavy chain and
humanized light chain of each of those antibodies is shown in Table
3.
TABLE-US-00003 TABLE 3 Humanized heavy chains and light chains of
huAb1 to huAb16 Humanized Humanized SEQ ID Humanized SEQ ID
antibody HC NO LC NO huAbl h0301-H0 53 h0301-L0 60 huAb2 h0301-H1
54 h0301-L0 60 huAb3 h0301-H2 55 h0301-L0 60 huAb4 h0301-H0 53
h0301-L1 61 huAb5 h0301-H1 54 h0301-L1 61 huAb6 h0301-H2 55
h0301-L1 61 huAb7 h0302-H1 56 h0302-L0 62 huAb8 h0302-H1 56
h0302-L1 63 huAb9 h0302-H1 56 h0302-L2 64 huAb10 h0302-H2 57
h0302-L0 62 huAb11 h0302-H2 57 h0302-L1 63 huAb12 h0302-H2 57
h0302-L2 64 huAb13 h0311-H1 58 h0311-L0 65 huAb14 h0311-H1 58
h0311-L1 66 huAb15 h0311-H2 59 h0311-L0 65 huAb16 h0311-H2 59
h0311-L1 66
[0247] The 16 humanized antibodies were tested for binding to
human, cynomolgus monkey, and mouse CSF1R ECD, as described
previously. See, e.g., PCT Publication No. WO 2011/140249. The
antibodies were found to bind to both human and cynomolgus monkey
CSF1R ECD, but not to mouse CSF1R ECD. The humanized antibodies
were also found to block binding of CSF1 and IL-34 to both human
and mouse CSF1R and to inhibit CSF1-induced and IL-34-induced
phosphorylation of human CSF1R expressed in CHO cells. See, e.g.,
PCT Publication No. WO 2011/140249.
[0248] The k.sub.a, k.sub.d, and K.sub.D for binding to human CSF1R
ECD were previously determined and are shown in Table 4. See, e.g.,
PCT Publication No. WO 2011/140249.
TABLE-US-00004 TABLE 4 Humanized antibody binding affinity for
human CSF1R huAb k.sub.a (M.sup.-1s.sup.-1) K.sub.d (s.sup.-1)
K.sub.D (nM) huAb 0301-L0H0 3.22 .times. 10.sup.6 1.11 .times.
10.sup.-03 0.35 huAb 0301-L0H1 3.56 .times. 10.sup.6 1.22 .times.
10.sup.-03 0.34 huAb 0301-L0H2 2.32 .times. 10.sup.6 6.60 .times.
10.sup.-04 0.28 huAb 0301-L1H0 3.29 .times. 10.sup.6 1.15 .times.
10.sup.-03 0.35 huAb 0301-L1H1 2.87 .times. 10.sup.6 9.21 .times.
10.sup.-04 0.32 huAb 0301-L1H2 2.95 .times. 10.sup.6 7.42 .times.
10.sup.-04 0.25 huAb 0302-L0H1 3.54 .times. 10.sup.6 3.69 .times.
10.sup.-03 1.04 huAb 0302-L1H1 3.47 .times. 10.sup.6 4.04 .times.
10.sup.-03 1.17 huAb 0302-L2H1 1.60 .times. 10.sup.6 9.14 .times.
10.sup.-04 0.57 huAb 0302-L0H2 3.40 .times. 10.sup.6 1.79 .times.
10.sup.-03 0.53 huAb 0302-L1H2 2.71 .times. 10.sup.6 1.53 .times.
10.sup.-03 0.56 huAb 0302-L2H2 1.84 .times. 10.sup.6 8.40 .times.
10.sup.-04 0.46 huAb 0311-L0H1 1.22 .times. 10.sup.6 5.40 .times.
10.sup.-04 0.44 huAb 0311-L1H1 1.32 .times. 10.sup.6 6.64 .times.
10.sup.-04 0.50 huAb 0311-L0H2 1.34 .times. 10.sup.6 4.73 .times.
10.sup.-04 0.35 huAb 0311-L1H2 1.51 .times. 10.sup.6 6.09 .times.
10.sup.-04 0.40
Example 2: HuAb1 Decreases CD16+ but not CD16- Peripheral Blood
Monocytes in Cynomolgus Monkeys
[0249] 0 mg/kg, 3 mg/kg, 10 mg/kg, or 30 mg/kg huAb1 was
administered intravenously to cynomolgus monkeys (2 monkeys per
group). Peripheral blood was drawn 14 days later, and CD16+ or
CD16- monocyte numbers were analyzed by flow cytometry. Briefly,
0.2 ml peripheral blood was labeled with 20 .mu.l anti-CD16-PE
(Becton-Dickenson) for 30 minutes at room temperature in the dark.
Erythrocytes were lysed by incubation with 2 ml of 1.times.BD
PharmLyse (Becton-Dickenson) for 20 min, and cells were washed with
1 ml FACS buffer (PBS/0.1% BSA) prior to analysis. Data was
collected on an LSRII flow cytometer using the Diva software
program (Becton-Dickenson), and results were analyzed using FloJo
data analysis program (Tree Star).
[0250] FIG. 3 shows that treatment of monkeys with huAb1 resulted
in a dose-dependent decrease in CD16+ but not CD16- peripheral
blood monocytes compared to untreated animals. These data show that
huAb1 can reduce monocytes that are mainly response for autoimmune
disorders while leaving other subsets of monocytes that are
responsible for pathogen clearance intact. This suggests that an
antibody that binds CSF1R and blocks ligand binding, such as huAb1,
may be effective for treating autoimmune disorders while
maintaining a good safety profile with respect to infections.
Example 3: Characterization of a Surrogate Anti-Mouse CSF1R
Antibody
[0251] Because humanized antibodies huAb1 to huAb16 do not bind to
mouse CSF1R, a chimeric anti-mouse CSF1R antibody ("cAb1") was
selected as a surrogate antibody for use in mouse models. The
antibody comprises rat variable regions and mouse constant regions.
To generate cAb1, rats were immunized with a mouse CSF1R-ECD-Fc
fusion protein, and hybridomas secreting antibodies that blocked
binding of both mouse CSF1 and mouse IL-34 to mouse CSF1R were
identified by ELISA assay. The variable regions of one such
blocking antibody were grafted onto mouse IgG constant regions
using standard molecular biologic techniques (Sambrook and Russell,
Molecular Cloning: A Laboratory Manual, Third Edition, CSH Press,
Cold Spring Harbor, N.Y., 2001). Recombinant chimeric antibody cAb1
was then expressed in CHO cells using standard techniques (Sambrook
and Russell, Ibid.).
[0252] Binding of cAb1 to mouse CSF1R was determined as follows.
Ninety-six well clear-bottom ELISA plates were coated overnight
with 1 .mu.g/ml recombinant mouse CSF1R ECD-Fc Chimera (R&D
Systems) in PBS. The next morning, wells were washed four times
with 0.05% Tween20 in PBS (PBST) and blocked with Blocker-Blotto
(Pierce). Fifty .mu.l of 0.5.times. serial dilutions of cAb1,
starting with a concentration of 2.6 .mu.g/ml, diluted 1:1 in
Blocker-Blotto were added to the CSF1R-coated wells. After
incubation at room temperature (RT) for 90 min, wells were washed
four times with PBST, and a 1:5000 dilution of a
peroxidase-conjugated goat anti-mouse IgG (Sigma) in Blocker-Blotto
was added to each well. After incubation at RT for 60 min, wells
were washed four times with PBST, and 50 .mu.l o-phenylenediamine
dihydrochloride peroxidase substrate (Sigma) was added to each
well. After incubation at RT for 30 min, A450 values of each well
were read directly on a SpectraMaxPlus spectrophotometer with
SoftMaxPro software (Molecular Devices).
[0253] The results of that experiment are shown in FIG. 4. cAb1
bound to mouse CSF1R ECD in a dose-dependent manner.
Example 4: cAb1 Blocks Ligand-Induced Proliferation of a Factor
Dependent Cell Line
[0254] cAb1 was tested for the ability to block ligand-induced
proliferation of the factor dependent mouse cell line mNFS60 as
follows. mNFS60 cells were stimulated with 10 ng/ml recombinant
mouse CSF1 or 100 ng/ml recombinant mouse IL-34 (both from R&D
Systems) in the presence or absence of serial dilutions of cAb1.
After incubation at 37.degree. C. for 48 hours, relative cellular
ATP content of each individual culture was assessed using Cell
Titer Glo reagent (Promega) according to the manufacturer's
instructions. In this assay, relative cellular ATP content is
directly proportional to the number of viable cells in culture, and
thus reflects the mNFS60 cell proliferation response.
[0255] The results of that experiment are shown in FIG. 5. cAb1
blocked CSF1 or IL-34 induced proliferation of mNFS60 cells in a
dose-dependent manner.
Example 5: cAb1 Suppresses Clinical Disease Scores in a Mouse Model
of Rheumatoid Arthritis
[0256] Arthritis was induced in male DBA/1 mice by intradermal
injection of 150 .mu.l of 2 mg/ml bovine Type II collagen (Elastin
Products, Owensville, Mo.) in Freund's complete adjuvant (Sigma) on
study day 0 and again on study day 21. Mice were randomized into
treatment groups on study day 0 and dosing was initiated. Mice were
dosed intraperitoneally 3 times per week (3.times./wk) with vehicle
or with cAb1 (3, 6, 10, or 30 mg/kg) through day 32 of the study,
and Clinical Disease Scores were measured for each paw on study
days 18 to 35. Clinical Disease Scores were assigned as follows:
[0257] 0=normal [0258] 1=1 hind or fore paw joint affected or
minimal diffuse erythema and swelling [0259] 2=2 hind or fore paw
joints affected or mild diffuse erythema and swelling [0260] 3=3
hind or fore paw joints affected or moderate diffuse erythema and
swelling [0261] 4=4 hind or fore paw joints affected or marked
diffuse erythema and swelling [0262] 5=Entire paw affected, severe
diffuse erythema and severe swelling, unable to flex digits
[0263] FIG. 6 shows the results of that experiment. cAb1 suppressed
Clinical Disease Scores in the Collagen Induced Arthritis model of
rheumatoid arthritis. These data suggest that in human patients, an
antibody that binds CSF1R and blocks ligand binding, such as huAb1,
may be effective for reducing the signs and symptoms of rheumatoid
arthritis.
Example 6: cAb1 Suppresses Bone Loss in a Mouse Model of Rheumatoid
Arthritis
[0264] High levels of Tartrate Resistant Acid Phosphatase 5b
(TRAP5b) are associated with bone remodeling, and have been
observed in various conditions characterized by increased bone
resorption. TRAP5b levels can therefore be used as a marker for
bone resorption (i.e., bone loss).
[0265] Arthritis was induced in mice, and mice were treated with
cAb1 as described in Example 5. Blood was collected at day 35, and
plasma levels of TRAP5b (Tartrate Resistant Acid Phosphatase 5b)
were measured using a commercial solid phase immunofixed enzyme
activity assay for the determination of osteoclast-derived TRAP5b
in mouse serum (Immunodiagnostics Systems) according to the
manufacturer's instructions. TRAP5b is a marker for bone
turnover.
[0266] FIG. 7 shows the results of that experiment. cAb1 suppressed
bone loss in the collagen induced arthritis mode of rheumatoid
arthritis, as measured by the suppression of the TRAP5b bone
turnover marker. These data suggest that in human patients, an
antibody that binds CSF1R and blocks ligand binding, such as huAb1,
may suppress bone erosions and joint destruction in rheumatoid
arthritis.
Example 7: cAb1 Suppresses Inflammation, Cartilage Damage, Pannus
Formation, and Bone Destruction in a Mouse Model of Rheumatoid
Arthritis
[0267] Arthritis was induced in mice, and mice were treated with
cAb1 as described in Example 5. At the end of the study, paws and
knees were collected and placed into neutral buffered formalin
(NBF; available from Sigma-Aldrich, cat #HT-501128) overnight and
then moved to 70% ethanol for histopathology and
immunohistochemistry.
[0268] Six joints from each animal were processed for
histopathologic evaluation. The joints were then scored as
follows:
[0269] Inflammation [0270] 0=Normal [0271] 1=Minimal infiltration
of inflammatory cells in synovium and periarticular tissue of
affected joints [0272] 2=Mild infiltration of inflammatory cells.
If referring to paws, generally restricted to affected joints (1-3
affected) [0273] 3=Moderate infiltration with moderate edema. If
referring to paws, restricted to affected joints, generally 3-4
joints, including at least one wrist or ankle joint [0274] 4=Marked
infiltration affecting most areas with marked edema, 1 or 2
unaffected joints may be present [0275] 5=Severe diffuse
infiltration with severe edema affecting all joints and
periarticular tissues
[0276] Pannus [0277] 0=Normal [0278] 1=Minimal infiltration of
pannus in cartilage and subchondral bone, marginal zones [0279]
2=Mild infiltration with marginal zone destruction of hard tissue
in affected joints [0280] 3=Moderate infiltration with moderate
hard tissue destruction in affected joints [0281] 4=Marked
infiltration with marked destruction of joint architecture,
affecting most joints [0282] 5=Severe infiltration associated with
total or near total destruction of joint architecture, affects all
joints
[0283] Cartilage Damage [0284] 0=Normal [0285] 1=Minimal=generally
minimal to mild loss of toluidine blue staining with no obvious
chondrocyte loss or collagen disruption in affected joints [0286]
2=Mild=generally mild loss of toluidine blue staining with focal
areas of chondrocyte loss and/or collagen disruption in some
affected joints [0287] 3=Moderate=generally moderate loss of
toluidine blue staining with multifocal chondrocyte loss and/or
collagen disruption in affected joints, some matrix remains on any
affected surface with areas of severe matrix loss [0288]
4=Marked=marked loss of toluidine blue staining with multifocal
marked (depth to deep zone) chondrocyte loss and/or collagen
disruption in most joints, if knee-one surface with total to near
total cartilage loss [0289] 5=Severe=severe diffuse loss of
toluidine blue staining with multifocal severe (depth to tide mark)
chondrocyte loss and/or collagen disruption in all joints, if
knee-2 or more surfaces with total to near total cartilage loss
[0290] Bone Resorption [0291] 0=Normal [0292] 1=Minimal=small areas
of resorption, not readily apparent on low magnification, rare
osteoclasts in affected joints, restricted to marginal zones [0293]
2=Mild=more numerous areas of resorption, not readily apparent on
low magnification, osteoclasts more numerous in affected joints,
restricted to marginal zones [0294] 3=Moderate=obvious resorption
of medullary trabecular and cortical bone without full thickness
defects in cortex, loss of some medullary trabeculae, lesion
apparent on low magnification, osteoclasts more numerous in
affected joints [0295] 4=Marked=Full thickness defects in cortical
bone, often with distortion of profile of remaining cortical
surface, marked loss of medullary bone, numerous osteoclasts,
affects most joints [0296] 5=Severe=Full thickness defects in
cortical bone and destruction of joint architecture of all joints A
sum total histologic score for each parameter for all six joints
was calculated.
[0297] FIG. 8 shows the results of that experiment. cAb1 suppressed
inflammation, cartilage damage, pannus formation, and bone
destruction in the Collagen Induced Arthritis model of rheumatoid
arthritis.
Example 8: cAb1 Suppresses Paw and Knee Joint Macrophage Numbers in
a Mouse Model of Rheumatoid Arthritis
[0298] Arthritis was induced in mice, and mice were treated with
cAb1 as described in Example 5, and paw and knee joint sections
were prepared as described in Example 7. Paw and knee joint
sections were stained with a peroxidase conjugated anti-F4/80
antibody, and macrophage numbers were enumerated by counting
positive staining cells under a microscope. Tissue sectioning and
staining were done by Histotox Labs, Boulder, Colo. F4/80 is a
mouse macrophage marker (Leenen et al., J. Immunol. Methods, 1994,
174: 5-19).
[0299] The results of that experiment are shown in FIG. 9. cAb1
reduced both paw and knee joint macrophage numbers in the Collagen
Induced Arthritis model of rheumatoid arthritis.
Example 9: cAb1 Suppresses Autoantibody Formation in a Mouse Model
of Rheumatoid Arthritis
[0300] Arthritis was induced in mice, and mice were treated with
cAb1 as described in Example 5. Blood was collected at day 35, and
plasma levels of anti-mouse collagen II antibodies were measured
using a mouse anti-type1 and type2 collagen IgG assay kit
(Chondrex) according to the manufacturer's instructions.
[0301] The results of that experiment are shown in FIG. 10. cAb1
suppressed the formation of anti-mouse collagen II autoantibodies
in the Collagen Induced Arthritis mode of rheumatoid arthritis.
These data suggest that in human patients, an antibody that binds
CSF1R and blocks ligand binding, such as huAb1, may reduce an
important trigger of joint inflammation, namely autoantibodies. In
addition, autoantibodies such as RF and ACPA may, in some
embodiments, be used to monitor drug effect.
Example 10: cAb1 Suppresses Bone Loss in a Mouse Model of
Established Rheumatoid Arthritis
[0302] Arthritis was induced in mice as described in Example 5.
Mice were treated intraperitoneally 3.times./week with 30 mg/kg
cAb1 starting when clinical disease scores reached 0.05 to 1.0.
Blood was collected 23 days after initial treatment with cAb1, and
plasma levels of TRAP5b (Tartrate Resistant Acid Phosphatase 5b)
were measured using a commercial solid phase immunofixed enzyme
activity assay for the determination of osteoclast-derived TRAP5b
in mouse serum (Immunodiagnostics Systems) according to the
manufacturer's instructions. TRAP5b is a marker for bone
turnover.
[0303] The results of that experiment are shown in FIG. 11. cAb1
administered after development of clinical disease scores
suppressed bone loss in the Collagen Induced Arthritis mode of
rheumatoid arthritis as measured by the suppression of the TRAP5b
bone turnover marker. These data suggest that in human patients, an
antibody that binds CSF1R and blocks ligand binding, such as huAb1,
may suppress bone erosion and joint destruction in rheumatoid
arthritis.
Example 11: cAb1 Suppresses Pannus Formation and Bone Destruction
in a Mouse Model of Established Rheumatoid Arthritis
[0304] Arthritis was induced in mice, and mice were treated with
cAb1 as described in Example 10. At the end of the study paws and
knees were collected and placed into neutral buffered formalin
(NBF) overnight and then moved to 70% ethanol for histopathology
and immunohistochemistry.
[0305] Six joints from each animal were processed for
histopathologic evaluation, and a sum total histologic score for
all six joints was calculated. The joints were scored as described
in Example 7.
[0306] The results of that experiment are shown in FIG. 12. cAb1
administered after development of clinical disease scores
suppressed pannus formation, and bone destruction in the Collagen
Induced Arthritis model of rheumatoid arthritis. Histologic scores
of inflammation and cartilage damage were unaffected. These data
also suggest that in human patients, an antibody that binds CSF1R
and blocks ligand binding, such as huAb1, may suppress bone erosion
and joint destruction in rheumatoid arthritis.
Example 12: cAb1 Suppresses Glomerulonephritis, Interstitial
Nephritis, and Perivascular Infiltrates in a Mouse Model of
Systemic Lupus Erythematosus
[0307] A subset of lupus patients develop lupus nephritis, which
involves inflammation in one or more structures in the kidney and
can lead to kidney failure. Lupus nephritis may detected in
patients and severity of disease assessed, for example, by
measuring proteinuria, or protein in the urine, and/or determining
the glomerular filtration rate, which is a measurement of how much
fluid is filtered through the kidneys per unit time. Lupus
nephritis may also be detected histologically through examination
of kidney biopsies, or through imaging techniques such as
ultrasonography (Hahn et al., Arthr. Care Res., 2012, 64:
797-808).
[0308] The MRL/MpJ-Fas.sup.lpr/J strain of mice spontaneously
develops lupus-like symptoms, including glomerular,
tubulointerstitial, and perivascular kidney disease;
lymphadenopathy; splenomegaly; circulating autoantibodies; and skin
lesions. Female MRL/MpJ-Fas.sup.lpr/J mice were dosed
intraperitoneally every day for 10 weeks from the age of 10 weeks
to the age of 20 weeks with vehicle or with 50 mg/kg cAb1. At the
end of the study (mouse age 20 weeks), kidneys were collected,
weighed, and placed in 10% neutral buffered formalin (NBF) for
Histopathologic evaluation. Kidneys were scored according to the
following system:
[0309] Glomerulonephritis [0310] 0=Normal. [0311] 1=Focal or
multifocal, minimal to mild, early proliferative. [0312]
2=Multifocal mild to moderate or mild diffuse proliferative. [0313]
3=Diffuse moderate proliferative with or without multifocal severe
areas. [0314] 4=Marked diffuse proliferative, with
crescents/sclerosis, multifocal protein casts present. [0315]
5=Severe diffuse proliferative, with crescents/sclerosis, diffuse
protein casts present.
[0316] Protein Cast Severity [0317] 0=Normal tubules. [0318]
1=Minimal=One to 5 profiles of tubules with protein casts. [0319]
2=Mild=Multifocal tubules with protein casts (6-20). [0320]
3=Moderate=Multifocal tubules with protein casts (21-40). [0321]
4=Marked=Multifocal tubules with protein casts (>40 but affects
less than [0322] 50% of all tubules). [0323] 5=Severe=Diffuse
tubules with protein casts, affects greater than 50% of all
Tubules.
[0324] Interstitial Nephritis (inflammation not obviously
associated with vessels) [0325] 0=Normal. [0326] 0.5=Very
Minimal=Small focal area of MNC in pelvis only. [0327]
1=Minimal=Occasional small focal, accumulations of MNC: affects
less than 10% of total interstitium and generally localized around
pelvis. [0328] 2=Mild=Multifocal small to larger infiltrates
distributed around pelvis and cortex: affects 10-25% of cortex
area. [0329] 3=Moderate=Multifocal small to extensive infiltrates
in pelvis and cortex: affects 26-50% of cortex area. [0330]
4=Marked=Multifocal to diffuse infiltration: affects 51-75% of
pelvis and cortex area. [0331] 5=Severe=Diffuse infiltration:
affects 76-100% of pelvis and cortex area.
[0332] Vessels [0333] 0=Normal. [0334] 0.5=Very Minimal=One vessel
with minimal perivascular infiltrate. [0335] 1=Minimal=Small but
definite perivascular infiltrates (1-2). [0336] 2=Mild=Several
(3-4) foci of perivascular infiltrate, no necrosis. [0337]
3=Moderate=Multifocal (5-6) foci of perivascular infiltrate, more
extensive, may have some necrosis of vessel wall. [0338]
4=Marked=Multifocal (7-8) foci of perivascular infiltrate,
extensive with necrosis. [0339] 5=Severe=Multifocal (>8) foci of
perivascular infiltrate, extensive with necrosis.
[0340] The results of that experiment are shown in FIG. 13. cAb1
suppressed glomerulonephritis, intersitital nephritis, and
perivascular infiltrates in the MRL/lpr model of lupus. Histologic
scores of protein cast severity were unaffected. These data suggest
that in human patients, an antibody that binds CSF1R and blocks
ligand binding, such as huAb1, may reduce renal inflammation
associated with systemic lupus erythematosus and restore renal
function.
Example 13: cAb1 Suppresses Skin Lesions in a Mouse Model of
Systemic Lupus Erythematosus
[0341] Female MRL/MpJ-Fas.sup.lpr/J mice were dosed with cAb1
described in Example 12. Mice were scored for the presence and
severity of skin lesions from the age of 10 weeks through the age
of 20 weeks. Skin lesions were scored as follows: [0342] 0=none
[0343] 1=mild (snout and ears) [0344] 2=moderate (<2 cm snout,
ears, and intrascapular) [0345] 3=severe (>2 cm snout, ears, and
intrascapular)
[0346] The results of that experiment are shown in FIG. 14. cAb1
suppressed the incidence and severity of skin lesions in the
MRL/lpr model of lupus. These data suggest that in human patients,
an antibody that binds CSF1R and blocks ligand binding, such as
huAb1, may reduce the systemic manifestations of SLE, including
skin inflammation, rash, and discoid lesions.
Example 14: cAb1 Suppresses Clinical Disease Scores in a Mouse
Model of Multiple Sclerosis
[0347] Experimental autoimmune encephalitis (EAE) was induced in
female C57Bl/6 mice by subcutaneous injection of 300 ng myelin
oligodendrocyte glycoprotein peptide 35-55 (MOG 35-55,
Sigma-Aldrich, Cat #M4939) in complete Freund's adjuvant on study
day 0 and again on study day 7. To enhance the induction and
consistency of EAE, 500 ng pertussis toxin (List Biological
Laboratories, Cat #180) was administered intraperitoneally on days
0 and 2. EAE is a model of multiple sclerosis. Mice were randomized
into treatment groups on study day 0 and dosing was initiated. Mice
were dosed intraperitoneally 3 times per week (3.times./wk) with
vehicle or with 30 mg/kg cAb1 through day 15 of the study. Clinical
Disease Scores were measured on study days 6-15. Clinical Disease
Scores were assigned as follows: [0348] 0.0=No clinical signs
[0349] 0.5=Partially limp tail [0350] 1.0=Paralyzed tail [0351]
2.0=Loss in coordinated movement; hind limb paresis [0352] 2.5=One
hind limb paralyzed [0353] 3.0=Both hind limbs paralyzed [0354]
3.5=Hind limbs paralyzed; weakness in forelimbs [0355]
4.0=Forelimbs and hindlimbs paralyzed [0356] 5.0=Moribund
[0357] The results of that experiment are shown in FIG. 15. cAb1
suppressed Clinical Disease Scores in the Experimental Autoimmune
Encephalitis model of multiple sclerosis. These data suggest that
in human patients, an antibody that binds CSF1R and blocks ligand
binding, such as huAb1, may reduce inflammation in the CNS
associated with multiple sclerosis and may reverse paralysis caused
by MS-associated demyelination and neuronal damage.
Table of Sequences
[0358] Table 5 provides certain sequences discussed herein. All
polypeptide and antibody sequences are shown without leader
sequences, unless otherwise indicated.
TABLE-US-00005 TABLE 5 Sequences and Descriptions SEQ ID NO
Description Sequence 1 hCSF1R IPVIEPSVPE LVVKPGATVT LRCVGNGSVE
WDGPPSPHWT LYSDGSSSIL (full-length, STNNATFQNT GTYRCTEPGD
PLGGSAAIHL YVKDPARPWN VLAQEVVVFE no leader DQDALLPCLL TDPVLEAGVS
LVRVRGRPLM RHTNYSFSPW HGFTIHRAKF sequence) IQSQDYQCSA LMGGRKVMSI
SIRLKVQKVI PGPPALTLVP AELVRIRGEA AQIVCSASSV DVNFDVFLQH NNTKLAIPQQ
SDFHNNRYQK VLTLNLDQVD FQHAGNYSCV ASNVQGKHST SMFFRVVESA YLNLSSEQNL
IQEVTVGEGL NLKVMVEAYP GLQGFNWTYL GPFSDHQPEP KLANATTKDT YRHTFTLSLP
RLKPSEAGRY SFLARNPGGW RALTFELTLR YPPEVSVIWT FINGSGTLLC AASGYPQPNV
TWLQCSGHTD RCDEAQVLQV WDDPYPEVLS QEPFHKVTVQ SLLTVETLEH NQTYECRAHN
SVGSGSWAFI PISAGAHTHP PDEFLFTPVV VACMSIMALL LLLLLLLLYK YKQKPKYQVR
WKIIESYEGN SYTFIDPTQL PYNEKWEFPR NNLQFGKTLG AGAFGKVVEA TAFGLGKEDA
VLKVAVKMLK STAHADEKEA LMSELKIMSH LGQHENIVNL LGACTHGGPV LVITEYCCYG
DLLNFLRRKA EAMLGPSLSP GQDPEGGVDY KNIHLEKKYV RRDSGFSSQG VDTYVEMRPV
STSSNDSFSE QDLDKEDGRP LELRDLLHFS SQVAQGMAFL ASKNCIHRDV AARNVLLTNG
HVAKIGDFGL ARDIMNDSNY IVKGNARLPV KWMAPESIFD CVYTVQSDVW SYGILLWEIF
SLGLNPYPGI LVNSKFYKLV KDGYQMAQPA FAPKNIYSIM QACWALEPTH RPTFQQICSF
LQEQAQEDRR ERDYTNLPSS SRSGGSGSSS SELEEESSSE HLTCCEQGDI AQPLLQPNNY
QFC 2 hCSF1R MGPGVLLLLL VATAWHGQGI PVIEPSVPEL VVKPGATVTL RCVGNGSVEW
(full-length, + DGPPSPHWTL YSDGSSSILS TNNATFQNTG TYRCTEPGDP
LGGSAAIHLY leader VKDPARPWNV LAQEVVVFED QDALLPCLLT DPVLEAGVSL
VRVRGRPLMR sequence) HTNYSFSPWH GFTIHRAKFI QSQDYQCSAL MGGRKVMSIS
IRLKVQKVIP GPPALTLVPA ELVRIRGEAA QIVCSASSVD VNFDVFLQHN NTKLAIPQQS
DFHNNRYQKV LTLNLDQVDF QHAGNYSCVA SNVQGKHSTS MFFRVVESAY LNLSSEQNLI
QEVTVGEGLN LKVMVEAYPG LQGFNWTYLG PFSDHQPEPK LANATTKDTY RHTFTLSLPR
LKPSEAGRYS FLARNPGGWR ALTFELTLRY PPEVSVIWTF INGSGTLLCA ASGYPQPNVT
WLQCSGHTDR CDEAQVLQVW DDPYPEVLSQ EPFHKVTVQS LLTVETLEHN QTYECRAHNS
VGSGSWAFIP ISAGAHTHPP DEFLFTPVVV ACMSIMALLL LLLLLLLYKY KQKPKYQVRW
KIIESYEGNS YTFIDPTQLP YNEKWEFPRN NLQFGKTLGA GAFGKVVEAT AFGLGKEDAV
LKVAVKMLKS TAHADEKEAL MSELKIMSHL GQHENIVNLL GACTHGGPVL VITEYCCYGD
LLNFLRRKAE AMLGPSLSPG QDPEGGVDYK NIHLEKKYVR RDSGFSSQGV DTYVEMRPVS
TSSNDSFSEQ DLDKEDGRPL ELRDLLHFSS QVAQGMAFLA SKNCIHRDVA ARNVLLTNGH
VAKIGDFGLA RDIMNDSNYI VKGNARLPVK WMAPESIFDC VYTVQSDVWS YGILLWEIFS
LGLNPYPGIL VNSKFYKLVK DGYQMAQPAF APKNIYSIMQ ACWALEPTHR PTFQQICSFL
QEQAQEDRRE RDYTNLPSSS RSGGSGSSSS ELEEESSSEH LTCCEQGDIA QPLLQPNNYQ
FC 5 hCSF1R IPVIEPSVPE LVVKPGATVT LRCVGNGSVE WDGPPSPHWT LYSDGSSSIL
ECD.506 STNNATFQNT GTYRCTEPGD PLGGSAAIHL YVKDPARPWN VLAQEVVVFE
DQDALLPCLL TDPVLEAGVS LVRVRGRPLM RHTNYSFSPW HGFTIHRAKF IQSQDYQCSA
LMGGRKVMSI SIRLKVQKVI PGPPALTLVP AELVRIRGEA AQIVCSASSV DVNFDVFLQH
NNTKLAIPQQ SDFHNNRYQK VLTLNLDQVD FQHAGNYSCV ASNVQGKHST SMFFRVVESA
YLNLSSEQNL IQEVTVGEGL NLKVMVEAYP GLQGFNWTYL GPFSDHQPEP KLANATTKDT
YRHTFTLSLP RLKPSEAGRY SFLARNPGGW RALTFELTLR YPPEVSVIWT FINGSGTLLC
AASGYPQPNV TWLQCSGHTD RCDEAQVLQV WDDPYPEVLS QEPFHKVTVQ SLLTVETLEH
NQTYECRAHN SVGSGSWAFI PISAGAH 6 hCSF1R IPVIEPSVPE LVVKPGATVT
LRCVGNGSVE WDGPPSPHWT LYSDGSSSIL ECD.506-Fc STNNATFQNT GTYRCTEPGD
PLGGSAAIHL YVKDPARPWN VLAQEVVVFE DQDALLPCLL TDPVLEAGVS LVRVRGRPLM
RHTNYSFSPW HGFTIHRAKF IQSQDYQCSA LMGGRKVMSI SIRLKVQKVI PGPPALTLVP
AELVRIRGEA AQIVCSASSV DVNFDVFLQH NNTKLAIPQQ SDFHNNRYQK VLTLNLDQVD
FQHAGNYSCV ASNVQGKHST SMFFRVVESA YLNLSSEQNL IQEVTVGEGL NLKVMVEAYP
GLQGFNWTYL GPFSDHQPEP KLANATTKDT YRHTFTLSLP RLKPSEAGRY SFLARNPGGW
RALTFELTLR YPPEVSVIWT FINGSGTLLC AASGYPQPNV TWLQCSGHTD RCDEAQVLQV
WDDPYPEVLS QEPFHKVTVQ SLLTVETLEH NQTYECRAHN SVGSGSWAFI PISAGAHEPK
SSDKTHTCPP CPAPELLGGP SVFLFPPKPK DTLMISRTPE VTCVVVDVSH EDPEVKFNWY
VDGVEVHNAK TKPREEQYNS TYRVVSVLTV LHQDWLNGKE YKCKVSNKAL PAPIEKTISK
AKGQPREPQV YTLPPSRDEL TKNQVSLTCL VKGFYPSDIA VEWESNGQPE NNYKTTPPVL
DSDGSFFLYS KLTVDKSRWQ QGNVFSCSVM HEALHNHYTQ KSLSLSPGK 7 cynoCSF1R
MGPGVLLLLL VVTAWHGQGI PVIEPSGPEL VVKPGETVTL RCVGNGSVEW ECD (with
DGPISPHWTL YSDGPSSVLT TTNATFQNTR TYRCTEPGDP LGGSAAIHLY leader
VKDPARPWNV LAKEVVVFED QDALLPCLLT DPVLEAGVSL VRLRGRPLLR sequence)
HTNYSFSPWH GFTIHRAKFI QGQDYQCSAL MGSRKVMSIS IRLKVQKVIP GPPALTLVPA
ELVRIRGEAA QIVCSASNID VDFDVFLQHN TTKLAIPQRS DFHDNRYQKV LTLSLGQVDF
QHAGNYSCVA SNVQGKHSTS MFFRVVESAY LDLSSEQNLI QEVTVGEGLN LKVMVEAYPG
LQGFNWTYLG PFSDHQPEPK LANATTKDTY RHTFTLSLPR LKPSEAGRYS FLARNPGGWR
ALTFELTLRY PPEVSVIWTS INGSGTLLCA ASGYPQPNVT WLQCAGHTDR CDEAQVLQVW
VDPHPEVLSQ EPFQKVTVQS LLTAETLEHN QTYECRAHNS VGSGSWAFIP ISAGAR 8
cynoCSF1R MGPGVLLLLL VVTAWHGQGI PVIEPSGPEL VVKPGETVTL RCVGNGSVEW
ECD-Fc DGPISPHWTL YSDGPSSVLT TTNATFQNTR TYRCTEPGDP LGGSAAIHLY (with
leader VKDPARPWNV LAKEVVVFED QDALLPCLLT DPVLEAGVSL VRLRGRPLLR
sequence) HTNYSFSPWH GFTIHRAKFI QGQDYQCSAL MGSRKVMSIS IRLKVQKVIP
GPPALTLVPA ELVRIRGEAA QIVCSASNID VDFDVFLQHN TTKLAIPQRS DFHDNRYQKV
LTLSLGQVDF QHAGNYSCVA SNVQGKHSTS MFFRVVESAY LDLSSEQNLI QEVTVGEGLN
LKVMVEAYPG LQGFNWTYLG PFSDHQPEPK LANATTKDTY RHTFTLSLPR LKPSEAGRYS
FLARNPGGWR ALTFELTLRY PPEVSVIWTS INGSGTLLCA ASGYPQPNVT WLQCAGHTDR
CDEAQVLQVW VDPHPEVLSQ EPFQKVTVQS LLTAETLEHN QTYECRAHNS VGSGSWAFIP
ISAGARGSEP KSSDKTHTCP PCPAPELLGG PSVFLFPPKP KDTLMISRTP EVTCVVVDVS
HEDPEVKFNW YVDGVEVHNA KTKPREEQYN STYRVVSVLT VLHQDWLNGK EYKCKVSNKA
LPAPIEKTIS KAKGQPREPQ VYTLPPSRDE LTKNQVSLTC LVKGFYPSDI AVEWESNGQP
ENNYKTTPPV LDSDGSFFLY SKLTVDKSRW QQGNVFSCSV MHEALHNHYT QKSLSLSPGK 3
Light chain METDTLLLWV LLLWVPGSTG leader sequence 4 Heavy chain
MAVLGLLLCL VTFPSCVLS leader sequence 9 Fab 0301 EVQLQQSGPE
LVRPGASVKM SCKASGYTFT DNYMIWVKQS HGKSLEWIGD heavy chain INPYNGGTTF
NQKFKGKATL TVEKSSSTAY MQLNSLTSED SAVYYCARES variable PYFSNLYVMD
YWGQGTSVTV SS region 10 Fab 0301 NIVLTQSPAS LAVSLGQRAT ISCKASQSVD
YDGDNYMNWY QQKPGQPPKL light chain LIYAASNLES GIPARFSGSG SGTDFTLNIH
PVEEEDAATY YCHLSNEDLS variable TFGGGTKLEI K region 11 Fab 0302
EIQLQQSGPE LVKPGASVKM SCKASGYTFS DFNIHWVKQK PGQGLEWIGY heavy chain
INPYTDVTVY NEKFKGKATL TSDRSSSTAY MDLSSLTSED SAVYYCASYF variable
DGTFDYALDY WGQGTSITVS S region 12 Fab 0302 DVVVTQTPAS LAVSLGQRAT
ISCRASESVD NYGLSFMNWF QQKPGQPPKL light chain LIYTASNLES GIPARFSGGG
SRTDFTLTID PVEADDAATY FCQQSKELPW variable TFGGGTRLEI K region 13
Fab 0311 EIQLQQSGPD LMKPGASVKM SCKASGYIFT DYNMHWVKQN QGKSLEWMGE
heavy chain INPNNGVVVY NQKFKGTTTL TVDKSSSTAY MDLHSLTSED SAVYYCTRAL
variable YHSNFGWYFD SWGKGTTLTV SS region 14 Fab 0311 DIVLTQSPAS
LAVSLGQRAT ISCKASQSVD YDGDSHMNWY QQKPGQPPKL light chain LIYTASNLES
GIPARFSGSG SGADFTLTIH PVEEEDAATY YCQQGNEDPW variable TFGGGTRLEI K
region 15 0301 heavy GYTFTDNYMI chain CDR1 16 0301 heavy DINPYNGGTT
FNQKFKG chain CDR2 17 0301 heavy ESPYFSNLYV MDY chain CDR3 18 0301
light KASQSVDYDG DNYMN chain CDR1 19 0301 light AASNLES chain CDR2
20 0301 light HLSNEDLST chain CDR3 21 0302 heavy GYTFSDFNIH chain
CDR1 22 0302 heavy YINPYTDVTV YNEKFKG chain CDR2 23 0302 heavy
YFDGTFDYAL DY chain CDR3 24 0302 light RASESVDNYG LSFMN chain CDR1
25 0302 light TASNLES chain CDR2 26 0302 light QQSKELPWT chain CDR3
27 0311 heavy GYIFTDYNMH chain CDR1 28 0311 heavy EINPNNGVVV
YNQKFKG chain CDR2 29 0311 heavy ALYHSNFGWY FDS chain CDR3 30 0311
light KASQSVDYDG DSHMN chain CDR1 31 0311 light TASNLES chain CDR2
32 0311 light QQGNEDPWT chain CDR3 33 cAb 0301 EVQLQQSGPE
LVRPGASVKM SCKASGYTFT DNYMIWVKQS HGKSLEWIGD heavy chain INPYNGGTTF
NQKFKGKATL TVEKSSSTAY MQLNSLTSED SAVYYCARES PYFSNLYVMD YWGQGTSVTV
SSASTKGPSV FPLAPCSRST SESTAALGCL VKDYFPEPVT VSWNSGALTS GVHTFPAVLQ
SSGLYSLSSV VTVPSSSLGT KTYTCNVDHK PSNTKVDKRV ESKYGPPCPP CPAPEFLGGP
SVFLFPPKPK DTLMISRTPE VTCVVVDVSQ EDPEVQFNWY VDGVEVHNAK TKPREEQFNS
TYRVVSVLTV LHQDWLNGKE YKCKVSNKGL PSSIEKTISK AKGQPREPQV YTLPPSQEEM
TKNQVSLTCL VKGFYPSDIA VEWESNGQPE NNYKTTPPVL DSDGSFFLYS RLTVDKSRWQ
EGNVFSCSVM HEALHNHYTQ KSLSLSLGK 34 cAb 0301 NIVLTQSPAS LAVSLGQRAT
ISCKASQSVD YDGDNYMNWY QQKPGQPPKL light chain LIYAASNLES GIPARFSGSG
SGTDFTLNIH PVEEEDAATY YCHLSNEDLS TFGGGTKLEI KRTVAAPSVF IFPPSDEQLK
SGTASVVCLL NNFYPREAKV QWKVDNALQS GNSQESVTEQ DSKDSTYSLS STLTLSKADY
EKHKVYACEV THQGLSSPVT KSFNRGEC 35 cAb 0302 EIQLQQSGPE LVKPGASVKM
SCKASGYTFS DFNIHWVKQK PGQGLEWIGY heavy chain INPYTDVTVY NEKFKGKATL
TSDRSSSTAY MDLSSLTSED SAVYYCASYF DGTFDYALDY WGQGTSITVS SASTKGPSVF
PLAPCSRSTS ESTAALGCLV KDYFPEPVTV SWNSGALTSG VHTFPAVLQS SGLYSLSSVV
TVPSSSLGTK TYTCNVDHKP SNTKVDKRVE SKYGPPCPPC PAPEFLGGPS VFLFPPKPKD
TLMISRTPEV TCVVVDVSQE DPEVQFNWYV DGVEVHNAKT KPREEQFNST YRVVSVLTVL
HQDWLNGKEY KCKVSNKGLP SSIEKTISKA KGQPREPQVY TLPPSQEEMT KNQVSLTCLV
KGFYPSDIAV EWESNGQPEN NYKTTPPVLD SDGSFFLYSR LTVDKSRWQE GNVFSCSVMH
EALHNHYTQK SLSLSLGK 36 cAb 0302 DVVVTQTPAS LAVSLGQRAT ISCRASESVD
NYGLSFMNWF QQKPGQPPKL light chain LIYTASNLES GIPARFSGGG SRTDFTLTID
PVEADDAATY FCQQSKELPW TFGGGTRLEI KRTVAAPSVF IFPPSDEQLK SGTASVVCLL
NNFYPREAKV QWKVDNALQS GNSQESVTEQ DSKDSTYSLS STLTLSKADY EKHKVYACEV
THQGLSSPVT KSFNRGEC 37 cAb 0311 EIQLQQSGPD LMKPGASVKM SCKASGYIFT
DYNMHWVKQN QGKSLEWMGE heavy chain INPNNGVVVY NQKFKGTTTL TVDKSSSTAY
MDLHSLTSED SAVYYCTRAL YHSNFGWYFD SWGKGTTLTV SSASTKGPSV FPLAPCSRST
SESTAALGCL VKDYFPEPVT VSWNSGALTS GVHTFPAVLQ SSGLYSLSSV VTVPSSSLGT
KTYTCNVDHK PSNTKVDKRV ESKYGPPCPP CPAPEFLGGP SVFLFPPKPK DTLMISRTPE
VTCVVVDVSQ EDPEVQFNWY VDGVEVHNAK TKPREEQFNS TYRVVSVLTV LHQDWLNGKE
YKCKVSNKGL PSSIEKTISK AKGQPREPQV YTLPPSQEEM TKNQVSLTCL VKGFYPSDIA
VEWESNGQPE NNYKTTPPVL DSDGSFFLYS RLTVDKSRWQ EGNVFSCSVM HEALHNHYTQ
KSLSLSLGK 38 cAb 0311 DIVLTQSPAS LAVSLGQRAT ISCKASQSVD YDGDSHMNWY
QQKPGQPPKL light chain LIYTASNLES GIPARFSGSG SGADFTLTIH PVEEEDAATY
YCQQGNEDPW TFGGGTRLEI KRTVAAPSVF IFPPSDEQLK SGTASVVCLL NNFYPREAKV
QWKVDNALQS GNSQESVTEQ DSKDSTYSLS STLTLSKADY EKHKVYACEV THQGLSSPVT
KSFNRGEC 39 h0301-H0 QVQLVQSGAE VKKPGSSVKV SCKASGYTFT DNYMIWVRQA
PGQGLEWMGD heavy chain INPYNGGTTF NQKFKGRVTI TADKSTSTAY MELSSLRSED
TAVYYCARES variable PYFSNLYVMD YWGQGTLVTV SS region
40 h0301-H1 QVQLVQSGAE VKKPGSSVKV SCKASGYTFT DNYMIWVRQA PGQGLEWMGD
heavy chain INPYNGGTTF NQKFKGRVTI TVDKSTSTAY MELSSLRSED TAVYYCARES
variable PYFSNLYVMD YWGQGTLVTV SS region 41 h0301-H2 QVQLVQSGAE
VKKPGSSVKV SCKASGYTFT DNYMIWVRQA PGQGLEWIGD heavy chain INPYNGGTTF
NQKFKGRATL TVDKSTSTAY MELSSLRSED TAVYYCARES variable PYFSNLYVMD
YWGQGTLVTV SS region 42 H0302-H1 QVQLVQSGAE VKKPGSSVKV SCKASGYTFS
DFNIHWVRQA PGQGLEWMGY heavy chain INPYTDVTVY NEKFKGRVTI TSDKSTSTAY
MELSSLRSED TAVYYCASYF variable DGTFDYALDY WGQGTLVTVS S region 43
H0302-H2 QVQLVQSGAE VKKPGSSVKV SCKASGYTFS DFNIHWVRQA PGQGLEWIGY
heavy chain INPYTDVTVY NEKFKGRATL TSDKSTSTAY MELSSLRSED TAVYYCASYF
variable DGTFDYALDY WGQGTLVTVS S region 44 H0311-H1 QVQLVQSGAE
VKKPGSSVKV SCKASGYIFT DYNMHWVRQA PGQGLEWMGE heavy chain INPNNGVVVY
NQKFKGRVTI TVDKSTSTAY MELSSLRSED TAVYYCTRAL variable YHSNFGWYFD
SWGQGTLVTV SS region 45 H0311-H2 QVQLVQSGAE VKKPGSSVKV SCKASGYIFT
DYNMHWVRQA PGQGLEWMGE heavy chain INPNNGVVVY NQKFKGTTTL TVDKSTSTAY
MELSSLRSED TAVYYCTRAL variable YHSNFGWYFD SWGQGTLVTV SS region 46
h0301-L0 EIVLTQSPAT LSLSPGERAT LSCKASQSVD YDGDNYMNWY QQKPGQAPRL
light chain LIYAASNLES GIPARFSGSG SGTDFTLTIS SLEPEDFAVY YCHLSNEDLS
variable TFGGGTKVEI K region 47 h0301-L1 NIVLTQSPAT LSLSPGERAT
LSCKASQSVD YDGDNYMNWY QQKPGQAPRL light chain LIYAASNLES GIPARFSGSG
SGTDFTLTIS SLEPEDFAVY YCHLSNEDLS variable TFGGGTKVEI K region 48
H0302-L0 EIVLTQSPAT LSLSPGERAT LSCRASESVD NYGLSFMNWY QQKPGQAPRL
light chain LIYTASNLES GIPARFSGSG SGTDFTLTIS SLEPEDFAVY YCQQSKELPW
variable TFGQGTKVEI K region 49 H0302-L1 EIVLTQSPAT LSLSPGERAT
LSCRASESVD NYGLSFMNWY QQKPGQAPRL light chain LIYTASNLES GIPARFSGSG
SRTDFTLTIS SLEPEDFAVY YCQQSKELPW variable TFGQGTKVEI K region 50
H0302-L2 EIVVTQSPAT LSLSPGERAT LSCRASESVD NYGLSFMNWF QQKPGQAPRL
light chain LIYTASNLES GIPARFSGSG SRTDFTLTIS SLEPEDFAVY YCQQSKELPW
variable TFGQGTKVEI K region 51 H0311-L0 EIVLTQSPAT LSLSPGERAT
LSCKASQSVD YDGDSHMNWY QQKPGQAPRL light chain LIYTASNLES GIPARFSGSG
SGTDFTLTIS SLEPEDFAVY YCQQGNEDPW variable TFGQGTKVEI K region 52
H0311-L1 DIVLTQSPAT LSLSPGERAT LSCKASQSVD YDGDSHMNWY QQKPGQAPRL
light chain LIYTASNLES GIPARFSGSG SGADFTLTIS SLEPEDFAVY YCQQGNEDPW
variable TFGQGTKVEI K region 53 h0301-H0 QVQLVQSGAE VKKPGSSVKV
SCKASGYTFT DNYMIWVRQA PGQGLEWMGD heavy chain INPYNGGTTF NQKFKGRVTI
TADKSTSTAY MELSSLRSED TAVYYCARES PYFSNLYVMD YWGQGTLVTV SSASTKGPSV
FPLAPCSRST SESTAALGCL VKDYFPEPVT VSWNSGALTS GVHTFPAVLQ SSGLYSLSSV
VTVPSSSLGT KTYTCNVDHK PSNTKVDKRV ESKYGPPCPP CPAPEFLGGP SVFLFPPKPK
DTLMISRTPE VTCVVVDVSQ EDPEVQFNWY VDGVEVHNAK TKPREEQFNS TYRVVSVLTV
LHQDWLNGKE YKCKVSNKGL PSSIEKTISK AKGQPREPQV YTLPPSQEEM TKNQVSLTCL
VKGFYPSDIA VEWESNGQPE NNYKTTPPVL DSDGSFFLYS RLTVDKSRWQ EGNVFSCSVM
HEALHNHYTQ KSLSLSLGK 54 h0301-H1 QVQLVQSGAE VKKPGSSVKV SCKASGYTFT
DNYMIWVRQA PGQGLEWMGD heavy chain INPYNGGTTF NQKFKGRVTI TVDKSTSTAY
MELSSLRSED TAVYYCARES PYFSNLYVMD YWGQGTLVTV SSASTKGPSV FPLAPCSRST
SESTAALGCL VKDYFPEPVT VSWNSGALTS GVHTFPAVLQ SSGLYSLSSV VTVPSSSLGT
KTYTCNVDHK PSNTKVDKRV ESKYGPPCPP CPAPEFLGGP SVFLFPPKPK DTLMISRTPE
VTCVVVDVSQ EDPEVQFNWY VDGVEVHNAK TKPREEQFNS TYRVVSVLTV LHQDWLNGKE
YKCKVSNKGL PSSIEKTISK AKGQPREPQV YTLPPSQEEM TKNQVSLTCL VKGFYPSDIA
VEWESNGQPE NNYKTTPPVL DSDGSFFLYS RLTVDKSRWQ EGNVFSCSVM HEALHNHYTQ
KSLSLSLGK 55 h0301-H2 QVQLVQSGAE VKKPGSSVKV SCKASGYTFT DNYMIWVRQA
PGQGLEWIGD heavy chain INPYNGGTTF NQKFKGRATL TVDKSTSTAY MELSSLRSED
TAVYYCARES PYFSNLYVMD YWGQGTLVTV SSASTKGPSV FPLAPCSRST SESTAALGCL
VKDYFPEPVT VSWNSGALTS GVHTFPAVLQ SSGLYSLSSV VTVPSSSLGT KTYTCNVDHK
PSNTKVDKRV ESKYGPPCPP CPAPEFLGGP SVFLFPPKPK DTLMISRTPE VTCVVVDVSQ
EDPEVQFNWY VDGVEVHNAK TKPREEQFNS TYRVVSVLTV LHQDWLNGKE YKCKVSNKGL
PSSIEKTISK AKGQPREPQV YTLPPSQEEM TKNQVSLTCL VKGFYPSDIA VEWESNGQPE
NNYKTTPPVL DSDGSFFLYS RLTVDKSRWQ EGNVFSCSVM HEALHNHYTQ KSLSLSLGK 56
H0302-H1 QVQLVQSGAE VKKPGSSVKV SCKASGYTFS DFNIHWVRQA PGQGLEWMGY
heavy chain INPYTDVTVY NEKFKGRVTI TSDKSTSTAY MELSSLRSED TAVYYCASYF
DGTFDYALDY WGQGTLVTVS SASTKGPSVF PLAPCSRSTS ESTAALGCLV KDYFPEPVTV
SWNSGALTSG VHTFPAVLQS SGLYSLSSVV TVPSSSLGTK TYTCNVDHKP SNTKVDKRVE
SKYGPPCPPC PAPEFLGGPS VFLFPPKPKD TLMISRTPEV TCVVVDVSQE DPEVQFNWYV
DGVEVHNAKT KPREEQFNST YRVVSVLTVL HQDWLNGKEY KCKVSNKGLP SSIEKTISKA
KGQPREPQVY TLPPSQEEMT KNQVSLTCLV KGFYPSDIAV EWESNGQPEN NYKTTPPVLD
SDGSFFLYSR LTVDKSRWQE GNVFSCSVMH EALHNHYTQK SLSLSLGK 57 H0302-H2
QVQLVQSGAE VKKPGSSVKV SCKASGYTFS DFNIHWVRQA PGQGLEWIGY heavy chain
INPYTDVTVY NEKFKGRATL TSDKSTSTAY MELSSLRSED TAVYYCASYF DGTFDYALDY
WGQGTLVTVS SASTKGPSVF PLAPCSRSTS ESTAALGCLV KDYFPEPVTV SWNSGALTSG
VHTFPAVLQS SGLYSLSSVV TVPSSSLGTK TYTCNVDHKP SNTKVDKRVE SKYGPPCPPC
PAPEFLGGPS VFLFPPKPKD TLMISRTPEV TCVVVDVSQE DPEVQFNWYV DGVEVHNAKT
KPREEQFNST YRVVSVLTVL HQDWLNGKEY KCKVSNKGLP SSIEKTISKA KGQPREPQVY
TLPPSQEEMT KNQVSLTCLV KGFYPSDIAV EWESNGQPEN NYKTTPPVLD SDGSFFLYSR
LTVDKSRWQE GNVFSCSVMH EALHNHYTQK SLSLSLGK 58 H0311-H1 QVQLVQSGAE
VKKPGSSVKV SCKASGYIFT DYNMHWVRQA PGQGLEWMGE heavy chain INPNNGVVVY
NQKFKGRVTI TVDKSTSTAY MELSSLRSED TAVYYCTRAL YHSNFGWYFD SWGQGTLVTV
SSASTKGPSV FPLAPCSRST SESTAALGCL VKDYFPEPVT VSWNSGALTS GVHTFPAVLQ
SSGLYSLSSV VTVPSSSLGT KTYTCNVDHK PSNTKVDKRV ESKYGPPCPP CPAPEFLGGP
SVFLFPPKPK DTLMISRTPE VTCVVVDVSQ EDPEVQFNWY VDGVEVHNAK TKPREEQFNS
TYRVVSVLTV LHQDWLNGKE YKCKVSNKGL PSSIEKTISK AKGQPREPQV YTLPPSQEEM
TKNQVSLTCL VKGFYPSDIA VEWESNGQPE NNYKTTPPVL DSDGSFFLYS RLTVDKSRWQ
EGNVFSCSVM HEALHNHYTQ KSLSLSLGK 59 H0311-H2 QVQLVQSGAE VKKPGSSVKV
SCKASGYIFT DYNMHWVRQA PGQGLEWMGE heavy chain INPNNGVVVY NQKFKGTTTL
TVDKSTSTAY MELSSLRSED TAVYYCTRAL YHSNFGWYFD SWGQGTLVTV SSASTKGPSV
FPLAPCSRST SESTAALGCL VKDYFPEPVT VSWNSGALTS GVHTFPAVLQ SSGLYSLSSV
VTVPSSSLGT KTYTCNVDHK PSNTKVDKRV ESKYGPPCPP CPAPEFLGGP SVFLFPPKPK
DTLMISRTPE VTCVVVDVSQ EDPEVQFNWY VDGVEVHNAK TKPREEQFNS TYRVVSVLTV
LHQDWLNGKE YKCKVSNKGL PSSIEKTISK AKGQPREPQV YTLPPSQEEM TKNQVSLTCL
VKGFYPSDIA VEWESNGQPE NNYKTTPPVL DSDGSFFLYS RLTVDKSRWQ EGNVFSCSVM
HEALHNHYTQ KSLSLSLGK 60 h0301-L0 EIVLTQSPAT LSLSPGERAT LSCKASQSVD
YDGDNYMNWY QQKPGQAPRL light chain LIYAASNLES GIPARFSGSG SGTDFTLTIS
SLEPEDFAVY YCHLSNEDLS TFGGGTKVEI KRTVAAPSVF IFPPSDEQLK SGTASVVCLL
NNFYPREAKV QWKVDNALQS GNSQESVTEQ DSKDSTYSLS STLTLSKADY EKHKVYACEV
THQGLSSPVT KSFNRGEC 61 h0301-L1 NIVLTQSPAT LSLSPGERAT LSCKASQSVD
YDGDNYMNWY QQKPGQAPRL light chain LIYAASNLES GIPARFSGSG SGTDFTLTIS
SLEPEDFAVY YCHLSNEDLS TFGGGTKVEI KRTVAAPSVF IFPPSDEQLK SGTASVVCLL
NNFYPREAKV QWKVDNALQS GNSQESVTEQ DSKDSTYSLS STLTLSKADY EKHKVYACEV
THQGLSSPVT KSFNRGEC 62 H0302-L0 EIVLTQSPAT LSLSPGERAT LSCRASESVD
NYGLSFMNWY QQKPGQAPRL light chain LIYTASNLES GIPARFSGSG SGTDFTLTIS
SLEPEDFAVY YCQQSKELPW TFGQGTKVEI KRTVAAPSVF IFPPSDEQLK SGTASVVCLL
NNFYPREAKV QWKVDNALQS GNSQESVTEQ DSKDSTYSLS STLTLSKADY EKHKVYACEV
THQGLSSPVT KSFNRGEC 63 H0302-L1 EIVLTQSPAT LSLSPGERAT LSCRASESVD
NYGLSFMNWY QQKPGQAPRL light chain LIYTASNLES GIPARFSGSG SRTDFTLTIS
SLEPEDFAVY YCQQSKELPW TFGQGTKVEI KRTVAAPSVF IFPPSDEQLK SGTASVVCLL
NNFYPREAKV QWKVDNALQS GNSQESVTEQ DSKDSTYSLS STLTLSKADY EKHKVYACEV
THQGLSSPVT KSFNRGEC 64 H0302-L2 EIVVTQSPAT LSLSPGERAT LSCRASESVD
NYGLSFMNWF QQKPGQAPRL light chain LIYTASNLES GIPARFSGSG SRTDFTLTIS
SLEPEDFAVY YCQQSKELPW TFGQGTKVEI KRTVAAPSVF IFPPSDEQLK SGTASVVCLL
NNFYPREAKV QWKVDNALQS GNSQESVTEQ DSKDSTYSLS STLTLSKADY EKHKVYACEV
THQGLSSPVT KSFNRGEC 65 H0311-L0 EIVLTQSPAT LSLSPGERAT LSCKASQSVD
YDGDSHMNWY QQKPGQAPRL light chain LIYTASNLES GIPARFSGSG SGTDFTLTIS
SLEPEDFAVY YCQQGNEDPW TFGQGTKVEI KRTVAAPSVF IFPPSDEQLK SGTASVVCLL
NNFYPREAKV QWKVDNALQS GNSQESVTEQ DSKDSTYSLS STLTLSKADY EKHKVYACEV
THQGLSSPVT KSFNRGEC 66 H0311-L1 DIVLTQSPAT LSLSPGERAT LSCKASQSVD
YDGDSHMNWY QQKPGQAPRL light chain LIYTASNLES GIPARFSGSG SGADFTLTIS
SLEPEDFAVY YCQQGNEDPW TFGQGTKVEI KRTVAAPSVF IFPPSDEQLK SGTASVVCLL
NNFYPREAKV QWKVDNALQS GNSQESVTEQ DSKDSTYSLS STLTLSKADY EKHKVYACEV
THQGLSSPVT KSFNRGEC 67 Human EEVSEYCSHM IGSGHLQSLQ RLIDSQMETS
CQITFEFVDQ EQLKDPVCYL CSF1 KKAFLLVQDI MEDTMRFRDN TPNAIAIVQL
QELSLRLKSC FTKDYEEHDK ACVRTFYETP LQLLEKVKNV FNETKNLLDK DWNIFSKNCN
NSFAECSSQG HERQSEGS 68 Human IL- NEPLEMWPLT QNEECTVTGF LRDKLQYRSR
LQYMKHYFPI NYKISVPYEG 34 VFRIANVTRL QRAQVSEREL RYLWVLVSLSATESVQDVLL
EGHPSWKYLQ EVQTLLLNVQ QGLTDVEVSP KVESVLSLLN APGPNLKLVR PKALLDNCFR
VMELLYCSCC KQSSVLNWQD CEVPSPQSCS PEPSLQYAAT QLYPPPPWSP SSPPHSTGSV
RPVRAQGEGL LP 69 Human QVQLVQSGAE VKKPGSSVKV SCKAS acceptor A FR1
70 Human WVRQAPGQGL EWMG acceptor A FR2 71 Human RVTITADKST
STAYMELSSL RSEDTAVYYC AR acceptor A FR3 72 Human WGQGTLVTVS S
acceptor A FR4 73 Human QVQLVQSGAE VKKPGSSVKV SCKAS acceptor B FR1
74 Human WVRQAPGQGL EWMG acceptor B FR2 75 Human RVTITADKST
STAYMELSSL RSEDTAVYYC AR acceptor B FR3 76 Human WGQGTLVTVSS
acceptor B FR4 77 Human QVQLVQSGAE VKKPGSSVKV SCKAS acceptor C FR1
78 Human WVRQAPGQGL EWMG acceptor C FR2 79 Human RVTITADKST
STAYMELSSL RSEDTAVYYC AR acceptor C FR3 80 Human WGQGTLVTVS S
acceptor C FR4 81 Human EIVLTQSPAT LSLSPGERAT LSC acceptor D FR1 82
Human acceptor D WYQQKPGQAP RLLIY FR2 83 Human acceptor D
GIPARFSGSG SGTDFTLTIS SLEPEDFAVY YC FR3 84 Human acceptor D
FGGGTKVEIK FR4
85 Human EIVLTQSPAT LSLSPGERAT LSC acceptor E FR1 86 Human
WYQQKPGQAP RLLIY acceptor E FR2 87 Human GIPARFSGSG SGTDFTLTIS
SLEPEDFAVY YC acceptor E FR3 88 Human FGQGTKVEIK acceptor E FR4 89
Human EIVLTQSPAT LSLSPGERAT LSC acceptor F FR1 90 Human WYQQKPGQAP
RLLIY acceptor F FR2 91 Human GIPARFSGSG SGTDFTLTIS SLEPEDFAVY YC
acceptor F FR3 92 Human FGQGTKVEIK acceptor F FR4 93 mCSF1R
APVIEPSGPE LVVEPGETVT LRCVSNGSVE WDGPISPYWT LDPESPGSTL ECD-Fc
TTRNATFKNT GTYRCTELED PMAGSTTIHL YVKDPAHSWN LLAQEVTVVE GQEAVLPCLI
TDPALKDSVS LMREGGRQVL RKTVYFFSPW RGFIIRKAKV LDSNTYVCKT MVNGRESTST
GIWLKVNRVH PEPPQIKLEP SKLVRIRGEA AQIVCSATNA EVGFNVILKR GDTKLEIPLN
SDFQDNYYKK VRALSLNAVD FQDAGIYSCV ASNDVGTRTA TMNFQVVESA YLNLTSEQSL
LQEVSVGDSL ILTVHADAYP SIQHYNWTYL GPFFEDQRKL EFITQRAIYR YTFKLFLNRV
KASEAGQYFL MAQNKAGWNN LTFELTLRYP PEVSVTWMPV NGSDVLFCDV SGYPQPSVTW
MECRGHTDRC DEAQALQVWN DTHPEVLSQK PFDKVIIQSQ LPIGTLKHNM TYFCKTHNSV
GNSSQYFRAV SLGQSKQEPK SSDKTHTCPP CPAPELLGGP SVFLFPPKPK DTLMISRTPE
VTCVVVDVSH EDPEVKFNWY VDGVEVHNAK TKPREEQYNS TYRVVSVLTV LHQDWLNGKE
YKCKVSNKAL PAPIEKTISK AKGQPREPQV YTLPPSRDEL TKNQVSLTCL VKGFYPSDIA
VEWESNGQPE NNYKTTPPVL DSDGSFFLYS KLTVDKSRWQ QGNVFSCSVM HEALHNHYTQ
KSLSLSPGK 94 Human ASTKGPSVFP LAPCSRSTSE STAALGCLVK DYFPEPVTVS
WNSGALTSGV IgG4 S241P HTFPAVLQSS GLYSLSSVVT VPSSSLGTKT YTCNVDHKPS
NTKVDKRVES KYGPPCPPCP APEFLGGPSV FLFPPKPKDT LMISRTPEVT CVVVDVSQED
PEVQFNWYVD GVEVHNAKTK PREEQFNSTY RVVSVLTVLH QDWLNGKEYK CKVSNKGLPS
SIEKTISKAK GQPREPQVYT LPPSQEEMTK NQVSLTCLVK GFYPSDIAVE WESNGQPENN
YKTTPPVLDS DGSFFLYSRL TVDKSRWQEG NVFSCSVMHE ALHNHYTQKS LSLSLGK 95
Human Ig.kappa. RTVAAPSVFI FPPSDEQLKS GTASVVCLLN NFYPREAKVQ
WKVDNALQSG NSQESVTEQD SKDSTYSLSS TLTLSKADYE KHKVYACEVT HQGLSSPVTK
SFNRGEC
Sequence CWU 1
1
951953PRTHomo sapiensmisc_feature(1)..(953)hCSF1R (full-length, no
leader sequence) 1Ile Pro Val Ile Glu Pro Ser Val Pro Glu Leu Val
Val Lys Pro Gly1 5 10 15Ala Thr Val Thr Leu Arg Cys Val Gly Asn Gly
Ser Val Glu Trp Asp 20 25 30Gly Pro Pro Ser Pro His Trp Thr Leu Tyr
Ser Asp Gly Ser Ser Ser 35 40 45Ile Leu Ser Thr Asn Asn Ala Thr Phe
Gln Asn Thr Gly Thr Tyr Arg 50 55 60Cys Thr Glu Pro Gly Asp Pro Leu
Gly Gly Ser Ala Ala Ile His Leu65 70 75 80Tyr Val Lys Asp Pro Ala
Arg Pro Trp Asn Val Leu Ala Gln Glu Val 85 90 95Val Val Phe Glu Asp
Gln Asp Ala Leu Leu Pro Cys Leu Leu Thr Asp 100 105 110Pro Val Leu
Glu Ala Gly Val Ser Leu Val Arg Val Arg Gly Arg Pro 115 120 125Leu
Met Arg His Thr Asn Tyr Ser Phe Ser Pro Trp His Gly Phe Thr 130 135
140Ile His Arg Ala Lys Phe Ile Gln Ser Gln Asp Tyr Gln Cys Ser
Ala145 150 155 160Leu Met Gly Gly Arg Lys Val Met Ser Ile Ser Ile
Arg Leu Lys Val 165 170 175Gln Lys Val Ile Pro Gly Pro Pro Ala Leu
Thr Leu Val Pro Ala Glu 180 185 190Leu Val Arg Ile Arg Gly Glu Ala
Ala Gln Ile Val Cys Ser Ala Ser 195 200 205Ser Val Asp Val Asn Phe
Asp Val Phe Leu Gln His Asn Asn Thr Lys 210 215 220Leu Ala Ile Pro
Gln Gln Ser Asp Phe His Asn Asn Arg Tyr Gln Lys225 230 235 240Val
Leu Thr Leu Asn Leu Asp Gln Val Asp Phe Gln His Ala Gly Asn 245 250
255Tyr Ser Cys Val Ala Ser Asn Val Gln Gly Lys His Ser Thr Ser Met
260 265 270Phe Phe Arg Val Val Glu Ser Ala Tyr Leu Asn Leu Ser Ser
Glu Gln 275 280 285Asn Leu Ile Gln Glu Val Thr Val Gly Glu Gly Leu
Asn Leu Lys Val 290 295 300Met Val Glu Ala Tyr Pro Gly Leu Gln Gly
Phe Asn Trp Thr Tyr Leu305 310 315 320Gly Pro Phe Ser Asp His Gln
Pro Glu Pro Lys Leu Ala Asn Ala Thr 325 330 335Thr Lys Asp Thr Tyr
Arg His Thr Phe Thr Leu Ser Leu Pro Arg Leu 340 345 350Lys Pro Ser
Glu Ala Gly Arg Tyr Ser Phe Leu Ala Arg Asn Pro Gly 355 360 365Gly
Trp Arg Ala Leu Thr Phe Glu Leu Thr Leu Arg Tyr Pro Pro Glu 370 375
380Val Ser Val Ile Trp Thr Phe Ile Asn Gly Ser Gly Thr Leu Leu
Cys385 390 395 400Ala Ala Ser Gly Tyr Pro Gln Pro Asn Val Thr Trp
Leu Gln Cys Ser 405 410 415Gly His Thr Asp Arg Cys Asp Glu Ala Gln
Val Leu Gln Val Trp Asp 420 425 430Asp Pro Tyr Pro Glu Val Leu Ser
Gln Glu Pro Phe His Lys Val Thr 435 440 445Val Gln Ser Leu Leu Thr
Val Glu Thr Leu Glu His Asn Gln Thr Tyr 450 455 460Glu Cys Arg Ala
His Asn Ser Val Gly Ser Gly Ser Trp Ala Phe Ile465 470 475 480Pro
Ile Ser Ala Gly Ala His Thr His Pro Pro Asp Glu Phe Leu Phe 485 490
495Thr Pro Val Val Val Ala Cys Met Ser Ile Met Ala Leu Leu Leu Leu
500 505 510Leu Leu Leu Leu Leu Leu Tyr Lys Tyr Lys Gln Lys Pro Lys
Tyr Gln 515 520 525Val Arg Trp Lys Ile Ile Glu Ser Tyr Glu Gly Asn
Ser Tyr Thr Phe 530 535 540Ile Asp Pro Thr Gln Leu Pro Tyr Asn Glu
Lys Trp Glu Phe Pro Arg545 550 555 560Asn Asn Leu Gln Phe Gly Lys
Thr Leu Gly Ala Gly Ala Phe Gly Lys 565 570 575Val Val Glu Ala Thr
Ala Phe Gly Leu Gly Lys Glu Asp Ala Val Leu 580 585 590Lys Val Ala
Val Lys Met Leu Lys Ser Thr Ala His Ala Asp Glu Lys 595 600 605Glu
Ala Leu Met Ser Glu Leu Lys Ile Met Ser His Leu Gly Gln His 610 615
620Glu Asn Ile Val Asn Leu Leu Gly Ala Cys Thr His Gly Gly Pro
Val625 630 635 640Leu Val Ile Thr Glu Tyr Cys Cys Tyr Gly Asp Leu
Leu Asn Phe Leu 645 650 655Arg Arg Lys Ala Glu Ala Met Leu Gly Pro
Ser Leu Ser Pro Gly Gln 660 665 670Asp Pro Glu Gly Gly Val Asp Tyr
Lys Asn Ile His Leu Glu Lys Lys 675 680 685Tyr Val Arg Arg Asp Ser
Gly Phe Ser Ser Gln Gly Val Asp Thr Tyr 690 695 700Val Glu Met Arg
Pro Val Ser Thr Ser Ser Asn Asp Ser Phe Ser Glu705 710 715 720Gln
Asp Leu Asp Lys Glu Asp Gly Arg Pro Leu Glu Leu Arg Asp Leu 725 730
735Leu His Phe Ser Ser Gln Val Ala Gln Gly Met Ala Phe Leu Ala Ser
740 745 750Lys Asn Cys Ile His Arg Asp Val Ala Ala Arg Asn Val Leu
Leu Thr 755 760 765Asn Gly His Val Ala Lys Ile Gly Asp Phe Gly Leu
Ala Arg Asp Ile 770 775 780Met Asn Asp Ser Asn Tyr Ile Val Lys Gly
Asn Ala Arg Leu Pro Val785 790 795 800Lys Trp Met Ala Pro Glu Ser
Ile Phe Asp Cys Val Tyr Thr Val Gln 805 810 815Ser Asp Val Trp Ser
Tyr Gly Ile Leu Leu Trp Glu Ile Phe Ser Leu 820 825 830Gly Leu Asn
Pro Tyr Pro Gly Ile Leu Val Asn Ser Lys Phe Tyr Lys 835 840 845Leu
Val Lys Asp Gly Tyr Gln Met Ala Gln Pro Ala Phe Ala Pro Lys 850 855
860Asn Ile Tyr Ser Ile Met Gln Ala Cys Trp Ala Leu Glu Pro Thr
His865 870 875 880Arg Pro Thr Phe Gln Gln Ile Cys Ser Phe Leu Gln
Glu Gln Ala Gln 885 890 895Glu Asp Arg Arg Glu Arg Asp Tyr Thr Asn
Leu Pro Ser Ser Ser Arg 900 905 910Ser Gly Gly Ser Gly Ser Ser Ser
Ser Glu Leu Glu Glu Glu Ser Ser 915 920 925Ser Glu His Leu Thr Cys
Cys Glu Gln Gly Asp Ile Ala Gln Pro Leu 930 935 940Leu Gln Pro Asn
Asn Tyr Gln Phe Cys945 9502972PRTHomo
sapiensmisc_feature(1)..(972)hCSF1R (full-length, + leader
sequence) 2Met Gly Pro Gly Val Leu Leu Leu Leu Leu Val Ala Thr Ala
Trp His1 5 10 15Gly Gln Gly Ile Pro Val Ile Glu Pro Ser Val Pro Glu
Leu Val Val 20 25 30Lys Pro Gly Ala Thr Val Thr Leu Arg Cys Val Gly
Asn Gly Ser Val 35 40 45Glu Trp Asp Gly Pro Pro Ser Pro His Trp Thr
Leu Tyr Ser Asp Gly 50 55 60Ser Ser Ser Ile Leu Ser Thr Asn Asn Ala
Thr Phe Gln Asn Thr Gly65 70 75 80Thr Tyr Arg Cys Thr Glu Pro Gly
Asp Pro Leu Gly Gly Ser Ala Ala 85 90 95Ile His Leu Tyr Val Lys Asp
Pro Ala Arg Pro Trp Asn Val Leu Ala 100 105 110Gln Glu Val Val Val
Phe Glu Asp Gln Asp Ala Leu Leu Pro Cys Leu 115 120 125Leu Thr Asp
Pro Val Leu Glu Ala Gly Val Ser Leu Val Arg Val Arg 130 135 140Gly
Arg Pro Leu Met Arg His Thr Asn Tyr Ser Phe Ser Pro Trp His145 150
155 160Gly Phe Thr Ile His Arg Ala Lys Phe Ile Gln Ser Gln Asp Tyr
Gln 165 170 175Cys Ser Ala Leu Met Gly Gly Arg Lys Val Met Ser Ile
Ser Ile Arg 180 185 190Leu Lys Val Gln Lys Val Ile Pro Gly Pro Pro
Ala Leu Thr Leu Val 195 200 205Pro Ala Glu Leu Val Arg Ile Arg Gly
Glu Ala Ala Gln Ile Val Cys 210 215 220Ser Ala Ser Ser Val Asp Val
Asn Phe Asp Val Phe Leu Gln His Asn225 230 235 240Asn Thr Lys Leu
Ala Ile Pro Gln Gln Ser Asp Phe His Asn Asn Arg 245 250 255Tyr Gln
Lys Val Leu Thr Leu Asn Leu Asp Gln Val Asp Phe Gln His 260 265
270Ala Gly Asn Tyr Ser Cys Val Ala Ser Asn Val Gln Gly Lys His Ser
275 280 285Thr Ser Met Phe Phe Arg Val Val Glu Ser Ala Tyr Leu Asn
Leu Ser 290 295 300Ser Glu Gln Asn Leu Ile Gln Glu Val Thr Val Gly
Glu Gly Leu Asn305 310 315 320Leu Lys Val Met Val Glu Ala Tyr Pro
Gly Leu Gln Gly Phe Asn Trp 325 330 335Thr Tyr Leu Gly Pro Phe Ser
Asp His Gln Pro Glu Pro Lys Leu Ala 340 345 350Asn Ala Thr Thr Lys
Asp Thr Tyr Arg His Thr Phe Thr Leu Ser Leu 355 360 365Pro Arg Leu
Lys Pro Ser Glu Ala Gly Arg Tyr Ser Phe Leu Ala Arg 370 375 380Asn
Pro Gly Gly Trp Arg Ala Leu Thr Phe Glu Leu Thr Leu Arg Tyr385 390
395 400Pro Pro Glu Val Ser Val Ile Trp Thr Phe Ile Asn Gly Ser Gly
Thr 405 410 415Leu Leu Cys Ala Ala Ser Gly Tyr Pro Gln Pro Asn Val
Thr Trp Leu 420 425 430Gln Cys Ser Gly His Thr Asp Arg Cys Asp Glu
Ala Gln Val Leu Gln 435 440 445Val Trp Asp Asp Pro Tyr Pro Glu Val
Leu Ser Gln Glu Pro Phe His 450 455 460Lys Val Thr Val Gln Ser Leu
Leu Thr Val Glu Thr Leu Glu His Asn465 470 475 480Gln Thr Tyr Glu
Cys Arg Ala His Asn Ser Val Gly Ser Gly Ser Trp 485 490 495Ala Phe
Ile Pro Ile Ser Ala Gly Ala His Thr His Pro Pro Asp Glu 500 505
510Phe Leu Phe Thr Pro Val Val Val Ala Cys Met Ser Ile Met Ala Leu
515 520 525Leu Leu Leu Leu Leu Leu Leu Leu Leu Tyr Lys Tyr Lys Gln
Lys Pro 530 535 540Lys Tyr Gln Val Arg Trp Lys Ile Ile Glu Ser Tyr
Glu Gly Asn Ser545 550 555 560Tyr Thr Phe Ile Asp Pro Thr Gln Leu
Pro Tyr Asn Glu Lys Trp Glu 565 570 575Phe Pro Arg Asn Asn Leu Gln
Phe Gly Lys Thr Leu Gly Ala Gly Ala 580 585 590Phe Gly Lys Val Val
Glu Ala Thr Ala Phe Gly Leu Gly Lys Glu Asp 595 600 605Ala Val Leu
Lys Val Ala Val Lys Met Leu Lys Ser Thr Ala His Ala 610 615 620Asp
Glu Lys Glu Ala Leu Met Ser Glu Leu Lys Ile Met Ser His Leu625 630
635 640Gly Gln His Glu Asn Ile Val Asn Leu Leu Gly Ala Cys Thr His
Gly 645 650 655Gly Pro Val Leu Val Ile Thr Glu Tyr Cys Cys Tyr Gly
Asp Leu Leu 660 665 670Asn Phe Leu Arg Arg Lys Ala Glu Ala Met Leu
Gly Pro Ser Leu Ser 675 680 685Pro Gly Gln Asp Pro Glu Gly Gly Val
Asp Tyr Lys Asn Ile His Leu 690 695 700Glu Lys Lys Tyr Val Arg Arg
Asp Ser Gly Phe Ser Ser Gln Gly Val705 710 715 720Asp Thr Tyr Val
Glu Met Arg Pro Val Ser Thr Ser Ser Asn Asp Ser 725 730 735Phe Ser
Glu Gln Asp Leu Asp Lys Glu Asp Gly Arg Pro Leu Glu Leu 740 745
750Arg Asp Leu Leu His Phe Ser Ser Gln Val Ala Gln Gly Met Ala Phe
755 760 765Leu Ala Ser Lys Asn Cys Ile His Arg Asp Val Ala Ala Arg
Asn Val 770 775 780Leu Leu Thr Asn Gly His Val Ala Lys Ile Gly Asp
Phe Gly Leu Ala785 790 795 800Arg Asp Ile Met Asn Asp Ser Asn Tyr
Ile Val Lys Gly Asn Ala Arg 805 810 815Leu Pro Val Lys Trp Met Ala
Pro Glu Ser Ile Phe Asp Cys Val Tyr 820 825 830Thr Val Gln Ser Asp
Val Trp Ser Tyr Gly Ile Leu Leu Trp Glu Ile 835 840 845Phe Ser Leu
Gly Leu Asn Pro Tyr Pro Gly Ile Leu Val Asn Ser Lys 850 855 860Phe
Tyr Lys Leu Val Lys Asp Gly Tyr Gln Met Ala Gln Pro Ala Phe865 870
875 880Ala Pro Lys Asn Ile Tyr Ser Ile Met Gln Ala Cys Trp Ala Leu
Glu 885 890 895Pro Thr His Arg Pro Thr Phe Gln Gln Ile Cys Ser Phe
Leu Gln Glu 900 905 910Gln Ala Gln Glu Asp Arg Arg Glu Arg Asp Tyr
Thr Asn Leu Pro Ser 915 920 925Ser Ser Arg Ser Gly Gly Ser Gly Ser
Ser Ser Ser Glu Leu Glu Glu 930 935 940Glu Ser Ser Ser Glu His Leu
Thr Cys Cys Glu Gln Gly Asp Ile Ala945 950 955 960Gln Pro Leu Leu
Gln Pro Asn Asn Tyr Gln Phe Cys 965 970320PRTMus
musculusmisc_feature(1)..(20)Light chain leader sequence 3Met Glu
Thr Asp Thr Leu Leu Leu Trp Val Leu Leu Leu Trp Val Pro1 5 10 15Gly
Ser Thr Gly 20419PRTMus musculusmisc_feature(1)..(19)Heavy chain
leader sequence 4Met Ala Val Leu Gly Leu Leu Leu Cys Leu Val Thr
Phe Pro Ser Cys1 5 10 15Val Leu Ser5487PRTHomo
sapiensmisc_feature(1)..(487)hCSF1R ECD.506 5Ile Pro Val Ile Glu
Pro Ser Val Pro Glu Leu Val Val Lys Pro Gly1 5 10 15Ala Thr Val Thr
Leu Arg Cys Val Gly Asn Gly Ser Val Glu Trp Asp 20 25 30Gly Pro Pro
Ser Pro His Trp Thr Leu Tyr Ser Asp Gly Ser Ser Ser 35 40 45Ile Leu
Ser Thr Asn Asn Ala Thr Phe Gln Asn Thr Gly Thr Tyr Arg 50 55 60Cys
Thr Glu Pro Gly Asp Pro Leu Gly Gly Ser Ala Ala Ile His Leu65 70 75
80Tyr Val Lys Asp Pro Ala Arg Pro Trp Asn Val Leu Ala Gln Glu Val
85 90 95Val Val Phe Glu Asp Gln Asp Ala Leu Leu Pro Cys Leu Leu Thr
Asp 100 105 110Pro Val Leu Glu Ala Gly Val Ser Leu Val Arg Val Arg
Gly Arg Pro 115 120 125Leu Met Arg His Thr Asn Tyr Ser Phe Ser Pro
Trp His Gly Phe Thr 130 135 140Ile His Arg Ala Lys Phe Ile Gln Ser
Gln Asp Tyr Gln Cys Ser Ala145 150 155 160Leu Met Gly Gly Arg Lys
Val Met Ser Ile Ser Ile Arg Leu Lys Val 165 170 175Gln Lys Val Ile
Pro Gly Pro Pro Ala Leu Thr Leu Val Pro Ala Glu 180 185 190Leu Val
Arg Ile Arg Gly Glu Ala Ala Gln Ile Val Cys Ser Ala Ser 195 200
205Ser Val Asp Val Asn Phe Asp Val Phe Leu Gln His Asn Asn Thr Lys
210 215 220Leu Ala Ile Pro Gln Gln Ser Asp Phe His Asn Asn Arg Tyr
Gln Lys225 230 235 240Val Leu Thr Leu Asn Leu Asp Gln Val Asp Phe
Gln His Ala Gly Asn 245 250 255Tyr Ser Cys Val Ala Ser Asn Val Gln
Gly Lys His Ser Thr Ser Met 260 265 270Phe Phe Arg Val Val Glu Ser
Ala Tyr Leu Asn Leu Ser Ser Glu Gln 275 280 285Asn Leu Ile Gln Glu
Val Thr Val Gly Glu Gly Leu Asn Leu Lys Val 290 295 300Met Val Glu
Ala Tyr Pro Gly Leu Gln Gly Phe Asn Trp Thr Tyr Leu305 310 315
320Gly Pro Phe Ser Asp His Gln Pro Glu Pro Lys Leu Ala Asn Ala Thr
325 330 335Thr Lys Asp Thr Tyr Arg His Thr Phe Thr Leu Ser Leu Pro
Arg Leu 340 345 350Lys Pro Ser Glu Ala Gly Arg Tyr Ser Phe Leu Ala
Arg Asn Pro Gly 355 360 365Gly Trp Arg Ala Leu Thr Phe Glu Leu Thr
Leu Arg Tyr Pro Pro Glu 370 375 380Val Ser Val Ile Trp Thr Phe Ile
Asn Gly Ser Gly Thr Leu Leu Cys385 390 395 400Ala Ala Ser Gly Tyr
Pro Gln Pro Asn Val Thr Trp Leu Gln Cys Ser 405 410 415Gly His Thr
Asp Arg Cys Asp Glu Ala Gln Val Leu Gln Val Trp Asp 420 425 430Asp
Pro Tyr Pro Glu Val Leu Ser Gln Glu Pro Phe His Lys Val Thr 435 440
445Val Gln Ser Leu Leu Thr Val Glu Thr Leu Glu His Asn Gln Thr Tyr
450
455 460Glu Cys Arg Ala His Asn Ser Val Gly Ser Gly Ser Trp Ala Phe
Ile465 470 475 480Pro Ile Ser Ala Gly Ala His 4856719PRTHomo
sapiensmisc_feature(1)..(719)hCSF1R ECD.506-Fc 6Ile Pro Val Ile Glu
Pro Ser Val Pro Glu Leu Val Val Lys Pro Gly1 5 10 15Ala Thr Val Thr
Leu Arg Cys Val Gly Asn Gly Ser Val Glu Trp Asp 20 25 30Gly Pro Pro
Ser Pro His Trp Thr Leu Tyr Ser Asp Gly Ser Ser Ser 35 40 45Ile Leu
Ser Thr Asn Asn Ala Thr Phe Gln Asn Thr Gly Thr Tyr Arg 50 55 60Cys
Thr Glu Pro Gly Asp Pro Leu Gly Gly Ser Ala Ala Ile His Leu65 70 75
80Tyr Val Lys Asp Pro Ala Arg Pro Trp Asn Val Leu Ala Gln Glu Val
85 90 95Val Val Phe Glu Asp Gln Asp Ala Leu Leu Pro Cys Leu Leu Thr
Asp 100 105 110Pro Val Leu Glu Ala Gly Val Ser Leu Val Arg Val Arg
Gly Arg Pro 115 120 125Leu Met Arg His Thr Asn Tyr Ser Phe Ser Pro
Trp His Gly Phe Thr 130 135 140Ile His Arg Ala Lys Phe Ile Gln Ser
Gln Asp Tyr Gln Cys Ser Ala145 150 155 160Leu Met Gly Gly Arg Lys
Val Met Ser Ile Ser Ile Arg Leu Lys Val 165 170 175Gln Lys Val Ile
Pro Gly Pro Pro Ala Leu Thr Leu Val Pro Ala Glu 180 185 190Leu Val
Arg Ile Arg Gly Glu Ala Ala Gln Ile Val Cys Ser Ala Ser 195 200
205Ser Val Asp Val Asn Phe Asp Val Phe Leu Gln His Asn Asn Thr Lys
210 215 220Leu Ala Ile Pro Gln Gln Ser Asp Phe His Asn Asn Arg Tyr
Gln Lys225 230 235 240Val Leu Thr Leu Asn Leu Asp Gln Val Asp Phe
Gln His Ala Gly Asn 245 250 255Tyr Ser Cys Val Ala Ser Asn Val Gln
Gly Lys His Ser Thr Ser Met 260 265 270Phe Phe Arg Val Val Glu Ser
Ala Tyr Leu Asn Leu Ser Ser Glu Gln 275 280 285Asn Leu Ile Gln Glu
Val Thr Val Gly Glu Gly Leu Asn Leu Lys Val 290 295 300Met Val Glu
Ala Tyr Pro Gly Leu Gln Gly Phe Asn Trp Thr Tyr Leu305 310 315
320Gly Pro Phe Ser Asp His Gln Pro Glu Pro Lys Leu Ala Asn Ala Thr
325 330 335Thr Lys Asp Thr Tyr Arg His Thr Phe Thr Leu Ser Leu Pro
Arg Leu 340 345 350Lys Pro Ser Glu Ala Gly Arg Tyr Ser Phe Leu Ala
Arg Asn Pro Gly 355 360 365Gly Trp Arg Ala Leu Thr Phe Glu Leu Thr
Leu Arg Tyr Pro Pro Glu 370 375 380Val Ser Val Ile Trp Thr Phe Ile
Asn Gly Ser Gly Thr Leu Leu Cys385 390 395 400Ala Ala Ser Gly Tyr
Pro Gln Pro Asn Val Thr Trp Leu Gln Cys Ser 405 410 415Gly His Thr
Asp Arg Cys Asp Glu Ala Gln Val Leu Gln Val Trp Asp 420 425 430Asp
Pro Tyr Pro Glu Val Leu Ser Gln Glu Pro Phe His Lys Val Thr 435 440
445Val Gln Ser Leu Leu Thr Val Glu Thr Leu Glu His Asn Gln Thr Tyr
450 455 460Glu Cys Arg Ala His Asn Ser Val Gly Ser Gly Ser Trp Ala
Phe Ile465 470 475 480Pro Ile Ser Ala Gly Ala His Glu Pro Lys Ser
Ser Asp Lys Thr His 485 490 495Thr Cys Pro Pro Cys Pro Ala Pro Glu
Leu Leu Gly Gly Pro Ser Val 500 505 510Phe Leu Phe Pro Pro Lys Pro
Lys Asp Thr Leu Met Ile Ser Arg Thr 515 520 525Pro Glu Val Thr Cys
Val Val Val Asp Val Ser His Glu Asp Pro Glu 530 535 540Val Lys Phe
Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys545 550 555
560Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser
565 570 575Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu
Tyr Lys 580 585 590Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile
Glu Lys Thr Ile 595 600 605Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro
Gln Val Tyr Thr Leu Pro 610 615 620Pro Ser Arg Asp Glu Leu Thr Lys
Asn Gln Val Ser Leu Thr Cys Leu625 630 635 640Val Lys Gly Phe Tyr
Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn 645 650 655Gly Gln Pro
Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser 660 665 670Asp
Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg 675 680
685Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu
690 695 700His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly
Lys705 710 7157506PRTMacaca
cynomolgusmisc_feature(1)..(506)cynoCSF1R ECD (with leader
sequence) 7Met Gly Pro Gly Val Leu Leu Leu Leu Leu Val Val Thr Ala
Trp His1 5 10 15Gly Gln Gly Ile Pro Val Ile Glu Pro Ser Gly Pro Glu
Leu Val Val 20 25 30Lys Pro Gly Glu Thr Val Thr Leu Arg Cys Val Gly
Asn Gly Ser Val 35 40 45Glu Trp Asp Gly Pro Ile Ser Pro His Trp Thr
Leu Tyr Ser Asp Gly 50 55 60Pro Ser Ser Val Leu Thr Thr Thr Asn Ala
Thr Phe Gln Asn Thr Arg65 70 75 80Thr Tyr Arg Cys Thr Glu Pro Gly
Asp Pro Leu Gly Gly Ser Ala Ala 85 90 95Ile His Leu Tyr Val Lys Asp
Pro Ala Arg Pro Trp Asn Val Leu Ala 100 105 110Lys Glu Val Val Val
Phe Glu Asp Gln Asp Ala Leu Leu Pro Cys Leu 115 120 125Leu Thr Asp
Pro Val Leu Glu Ala Gly Val Ser Leu Val Arg Leu Arg 130 135 140Gly
Arg Pro Leu Leu Arg His Thr Asn Tyr Ser Phe Ser Pro Trp His145 150
155 160Gly Phe Thr Ile His Arg Ala Lys Phe Ile Gln Gly Gln Asp Tyr
Gln 165 170 175Cys Ser Ala Leu Met Gly Ser Arg Lys Val Met Ser Ile
Ser Ile Arg 180 185 190Leu Lys Val Gln Lys Val Ile Pro Gly Pro Pro
Ala Leu Thr Leu Val 195 200 205Pro Ala Glu Leu Val Arg Ile Arg Gly
Glu Ala Ala Gln Ile Val Cys 210 215 220Ser Ala Ser Asn Ile Asp Val
Asp Phe Asp Val Phe Leu Gln His Asn225 230 235 240Thr Thr Lys Leu
Ala Ile Pro Gln Arg Ser Asp Phe His Asp Asn Arg 245 250 255Tyr Gln
Lys Val Leu Thr Leu Ser Leu Gly Gln Val Asp Phe Gln His 260 265
270Ala Gly Asn Tyr Ser Cys Val Ala Ser Asn Val Gln Gly Lys His Ser
275 280 285Thr Ser Met Phe Phe Arg Val Val Glu Ser Ala Tyr Leu Asp
Leu Ser 290 295 300Ser Glu Gln Asn Leu Ile Gln Glu Val Thr Val Gly
Glu Gly Leu Asn305 310 315 320Leu Lys Val Met Val Glu Ala Tyr Pro
Gly Leu Gln Gly Phe Asn Trp 325 330 335Thr Tyr Leu Gly Pro Phe Ser
Asp His Gln Pro Glu Pro Lys Leu Ala 340 345 350Asn Ala Thr Thr Lys
Asp Thr Tyr Arg His Thr Phe Thr Leu Ser Leu 355 360 365Pro Arg Leu
Lys Pro Ser Glu Ala Gly Arg Tyr Ser Phe Leu Ala Arg 370 375 380Asn
Pro Gly Gly Trp Arg Ala Leu Thr Phe Glu Leu Thr Leu Arg Tyr385 390
395 400Pro Pro Glu Val Ser Val Ile Trp Thr Ser Ile Asn Gly Ser Gly
Thr 405 410 415Leu Leu Cys Ala Ala Ser Gly Tyr Pro Gln Pro Asn Val
Thr Trp Leu 420 425 430Gln Cys Ala Gly His Thr Asp Arg Cys Asp Glu
Ala Gln Val Leu Gln 435 440 445Val Trp Val Asp Pro His Pro Glu Val
Leu Ser Gln Glu Pro Phe Gln 450 455 460Lys Val Thr Val Gln Ser Leu
Leu Thr Ala Glu Thr Leu Glu His Asn465 470 475 480Gln Thr Tyr Glu
Cys Arg Ala His Asn Ser Val Gly Ser Gly Ser Trp 485 490 495Ala Phe
Ile Pro Ile Ser Ala Gly Ala Arg 500 5058740PRTMacaca
cynomolgusmisc_feature(1)..(740)cynoCSF1R ECD-Fc (with leader
sequence) 8Met Gly Pro Gly Val Leu Leu Leu Leu Leu Val Val Thr Ala
Trp His1 5 10 15Gly Gln Gly Ile Pro Val Ile Glu Pro Ser Gly Pro Glu
Leu Val Val 20 25 30Lys Pro Gly Glu Thr Val Thr Leu Arg Cys Val Gly
Asn Gly Ser Val 35 40 45Glu Trp Asp Gly Pro Ile Ser Pro His Trp Thr
Leu Tyr Ser Asp Gly 50 55 60Pro Ser Ser Val Leu Thr Thr Thr Asn Ala
Thr Phe Gln Asn Thr Arg65 70 75 80Thr Tyr Arg Cys Thr Glu Pro Gly
Asp Pro Leu Gly Gly Ser Ala Ala 85 90 95Ile His Leu Tyr Val Lys Asp
Pro Ala Arg Pro Trp Asn Val Leu Ala 100 105 110Lys Glu Val Val Val
Phe Glu Asp Gln Asp Ala Leu Leu Pro Cys Leu 115 120 125Leu Thr Asp
Pro Val Leu Glu Ala Gly Val Ser Leu Val Arg Leu Arg 130 135 140Gly
Arg Pro Leu Leu Arg His Thr Asn Tyr Ser Phe Ser Pro Trp His145 150
155 160Gly Phe Thr Ile His Arg Ala Lys Phe Ile Gln Gly Gln Asp Tyr
Gln 165 170 175Cys Ser Ala Leu Met Gly Ser Arg Lys Val Met Ser Ile
Ser Ile Arg 180 185 190Leu Lys Val Gln Lys Val Ile Pro Gly Pro Pro
Ala Leu Thr Leu Val 195 200 205Pro Ala Glu Leu Val Arg Ile Arg Gly
Glu Ala Ala Gln Ile Val Cys 210 215 220Ser Ala Ser Asn Ile Asp Val
Asp Phe Asp Val Phe Leu Gln His Asn225 230 235 240Thr Thr Lys Leu
Ala Ile Pro Gln Arg Ser Asp Phe His Asp Asn Arg 245 250 255Tyr Gln
Lys Val Leu Thr Leu Ser Leu Gly Gln Val Asp Phe Gln His 260 265
270Ala Gly Asn Tyr Ser Cys Val Ala Ser Asn Val Gln Gly Lys His Ser
275 280 285Thr Ser Met Phe Phe Arg Val Val Glu Ser Ala Tyr Leu Asp
Leu Ser 290 295 300Ser Glu Gln Asn Leu Ile Gln Glu Val Thr Val Gly
Glu Gly Leu Asn305 310 315 320Leu Lys Val Met Val Glu Ala Tyr Pro
Gly Leu Gln Gly Phe Asn Trp 325 330 335Thr Tyr Leu Gly Pro Phe Ser
Asp His Gln Pro Glu Pro Lys Leu Ala 340 345 350Asn Ala Thr Thr Lys
Asp Thr Tyr Arg His Thr Phe Thr Leu Ser Leu 355 360 365Pro Arg Leu
Lys Pro Ser Glu Ala Gly Arg Tyr Ser Phe Leu Ala Arg 370 375 380Asn
Pro Gly Gly Trp Arg Ala Leu Thr Phe Glu Leu Thr Leu Arg Tyr385 390
395 400Pro Pro Glu Val Ser Val Ile Trp Thr Ser Ile Asn Gly Ser Gly
Thr 405 410 415Leu Leu Cys Ala Ala Ser Gly Tyr Pro Gln Pro Asn Val
Thr Trp Leu 420 425 430Gln Cys Ala Gly His Thr Asp Arg Cys Asp Glu
Ala Gln Val Leu Gln 435 440 445Val Trp Val Asp Pro His Pro Glu Val
Leu Ser Gln Glu Pro Phe Gln 450 455 460Lys Val Thr Val Gln Ser Leu
Leu Thr Ala Glu Thr Leu Glu His Asn465 470 475 480Gln Thr Tyr Glu
Cys Arg Ala His Asn Ser Val Gly Ser Gly Ser Trp 485 490 495Ala Phe
Ile Pro Ile Ser Ala Gly Ala Arg Gly Ser Glu Pro Lys Ser 500 505
510Ser Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu
515 520 525Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp
Thr Leu 530 535 540Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val
Val Asp Val Ser545 550 555 560His Glu Asp Pro Glu Val Lys Phe Asn
Trp Tyr Val Asp Gly Val Glu 565 570 575Val His Asn Ala Lys Thr Lys
Pro Arg Glu Glu Gln Tyr Asn Ser Thr 580 585 590Tyr Arg Val Val Ser
Val Leu Thr Val Leu His Gln Asp Trp Leu Asn 595 600 605Gly Lys Glu
Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro 610 615 620Ile
Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln625 630
635 640Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln
Val 645 650 655Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp
Ile Ala Val 660 665 670Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn
Tyr Lys Thr Thr Pro 675 680 685Pro Val Leu Asp Ser Asp Gly Ser Phe
Phe Leu Tyr Ser Lys Leu Thr 690 695 700Val Asp Lys Ser Arg Trp Gln
Gln Gly Asn Val Phe Ser Cys Ser Val705 710 715 720Met His Glu Ala
Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu 725 730 735Ser Pro
Gly Lys 7409122PRTMus musculusmisc_feature(1)..(122)Fab 0301 heavy
chain variable region 9Glu Val Gln Leu Gln Gln Ser Gly Pro Glu Leu
Val Arg Pro Gly Ala1 5 10 15Ser Val Lys Met Ser Cys Lys Ala Ser Gly
Tyr Thr Phe Thr Asp Asn 20 25 30Tyr Met Ile Trp Val Lys Gln Ser His
Gly Lys Ser Leu Glu Trp Ile 35 40 45Gly Asp Ile Asn Pro Tyr Asn Gly
Gly Thr Thr Phe Asn Gln Lys Phe 50 55 60Lys Gly Lys Ala Thr Leu Thr
Val Glu Lys Ser Ser Ser Thr Ala Tyr65 70 75 80Met Gln Leu Asn Ser
Leu Thr Ser Glu Asp Ser Ala Val Tyr Tyr Cys 85 90 95Ala Arg Glu Ser
Pro Tyr Phe Ser Asn Leu Tyr Val Met Asp Tyr Trp 100 105 110Gly Gln
Gly Thr Ser Val Thr Val Ser Ser 115 12010111PRTMus
musculusmisc_feature(1)..(111)Fab 0301 light chain variable region
10Asn Ile Val Leu Thr Gln Ser Pro Ala Ser Leu Ala Val Ser Leu Gly1
5 10 15Gln Arg Ala Thr Ile Ser Cys Lys Ala Ser Gln Ser Val Asp Tyr
Asp 20 25 30Gly Asp Asn Tyr Met Asn Trp Tyr Gln Gln Lys Pro Gly Gln
Pro Pro 35 40 45Lys Leu Leu Ile Tyr Ala Ala Ser Asn Leu Glu Ser Gly
Ile Pro Ala 50 55 60Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr
Leu Asn Ile His65 70 75 80Pro Val Glu Glu Glu Asp Ala Ala Thr Tyr
Tyr Cys His Leu Ser Asn 85 90 95Glu Asp Leu Ser Thr Phe Gly Gly Gly
Thr Lys Leu Glu Ile Lys 100 105 11011121PRTMus
musculusmisc_feature(1)..(121)Fab 0302 heavy chain variable region
11Glu Ile Gln Leu Gln Gln Ser Gly Pro Glu Leu Val Lys Pro Gly Ala1
5 10 15Ser Val Lys Met Ser Cys Lys Ala Ser Gly Tyr Thr Phe Ser Asp
Phe 20 25 30Asn Ile His Trp Val Lys Gln Lys Pro Gly Gln Gly Leu Glu
Trp Ile 35 40 45Gly Tyr Ile Asn Pro Tyr Thr Asp Val Thr Val Tyr Asn
Glu Lys Phe 50 55 60Lys Gly Lys Ala Thr Leu Thr Ser Asp Arg Ser Ser
Ser Thr Ala Tyr65 70 75 80Met Asp Leu Ser Ser Leu Thr Ser Glu Asp
Ser Ala Val Tyr Tyr Cys 85 90 95Ala Ser Tyr Phe Asp Gly Thr Phe Asp
Tyr Ala Leu Asp Tyr Trp Gly 100 105 110Gln Gly Thr Ser Ile Thr Val
Ser Ser 115 12012111PRTMus musculusmisc_feature(1)..(111)Fab 0302
light chain variable region 12Asp Val Val Val Thr Gln Thr Pro Ala
Ser Leu Ala Val Ser Leu Gly1 5 10 15Gln Arg Ala Thr Ile Ser Cys Arg
Ala Ser Glu Ser Val Asp Asn Tyr 20 25 30Gly Leu Ser Phe Met Asn Trp
Phe Gln Gln Lys Pro Gly Gln
Pro Pro 35 40 45Lys Leu Leu Ile Tyr Thr Ala Ser Asn Leu Glu Ser Gly
Ile Pro Ala 50 55 60Arg Phe Ser Gly Gly Gly Ser Arg Thr Asp Phe Thr
Leu Thr Ile Asp65 70 75 80Pro Val Glu Ala Asp Asp Ala Ala Thr Tyr
Phe Cys Gln Gln Ser Lys 85 90 95Glu Leu Pro Trp Thr Phe Gly Gly Gly
Thr Arg Leu Glu Ile Lys 100 105 11013122PRTMus
musculusmisc_feature(1)..(122)Fab 0311 heavy chain variable region
13Glu Ile Gln Leu Gln Gln Ser Gly Pro Asp Leu Met Lys Pro Gly Ala1
5 10 15Ser Val Lys Met Ser Cys Lys Ala Ser Gly Tyr Ile Phe Thr Asp
Tyr 20 25 30Asn Met His Trp Val Lys Gln Asn Gln Gly Lys Ser Leu Glu
Trp Met 35 40 45Gly Glu Ile Asn Pro Asn Asn Gly Val Val Val Tyr Asn
Gln Lys Phe 50 55 60Lys Gly Thr Thr Thr Leu Thr Val Asp Lys Ser Ser
Ser Thr Ala Tyr65 70 75 80Met Asp Leu His Ser Leu Thr Ser Glu Asp
Ser Ala Val Tyr Tyr Cys 85 90 95Thr Arg Ala Leu Tyr His Ser Asn Phe
Gly Trp Tyr Phe Asp Ser Trp 100 105 110Gly Lys Gly Thr Thr Leu Thr
Val Ser Ser 115 12014111PRTMus musculusmisc_feature(1)..(111)Fab
0311 light chain variable region 14Asp Ile Val Leu Thr Gln Ser Pro
Ala Ser Leu Ala Val Ser Leu Gly1 5 10 15Gln Arg Ala Thr Ile Ser Cys
Lys Ala Ser Gln Ser Val Asp Tyr Asp 20 25 30Gly Asp Ser His Met Asn
Trp Tyr Gln Gln Lys Pro Gly Gln Pro Pro 35 40 45Lys Leu Leu Ile Tyr
Thr Ala Ser Asn Leu Glu Ser Gly Ile Pro Ala 50 55 60Arg Phe Ser Gly
Ser Gly Ser Gly Ala Asp Phe Thr Leu Thr Ile His65 70 75 80Pro Val
Glu Glu Glu Asp Ala Ala Thr Tyr Tyr Cys Gln Gln Gly Asn 85 90 95Glu
Asp Pro Trp Thr Phe Gly Gly Gly Thr Arg Leu Glu Ile Lys 100 105
1101510PRTMus musculusmisc_feature(1)..(10)0301 heavy chain CDR1
15Gly Tyr Thr Phe Thr Asp Asn Tyr Met Ile1 5 101617PRTMus
musculusmisc_feature(1)..(17)0301 heavy chain CDR2 16Asp Ile Asn
Pro Tyr Asn Gly Gly Thr Thr Phe Asn Gln Lys Phe Lys1 5 10
15Gly1713PRTMus musculusmisc_feature(1)..(13)0301 heavy chain CDR3
17Glu Ser Pro Tyr Phe Ser Asn Leu Tyr Val Met Asp Tyr1 5
101815PRTMus musculusmisc_feature(1)..(15)0301 light chain CDR1
18Lys Ala Ser Gln Ser Val Asp Tyr Asp Gly Asp Asn Tyr Met Asn1 5 10
15197PRTMus musculusmisc_feature(1)..(7)0301 light chain CDR2 19Ala
Ala Ser Asn Leu Glu Ser1 5209PRTMus
musculusmisc_feature(1)..(9)0301 light chain CDR3 20His Leu Ser Asn
Glu Asp Leu Ser Thr1 52110PRTMus musculusmisc_feature(1)..(10)0302
heavy chain CDR1 21Gly Tyr Thr Phe Ser Asp Phe Asn Ile His1 5
102217PRTMus musculusmisc_feature(1)..(17)0302 heavy chain CDR2
22Tyr Ile Asn Pro Tyr Thr Asp Val Thr Val Tyr Asn Glu Lys Phe Lys1
5 10 15Gly2312PRTMus musculusmisc_feature(1)..(12)0302 heavy chain
CDR3 23Tyr Phe Asp Gly Thr Phe Asp Tyr Ala Leu Asp Tyr1 5
102415PRTMus musculusmisc_feature(1)..(15)0302 light chain CDR1
24Arg Ala Ser Glu Ser Val Asp Asn Tyr Gly Leu Ser Phe Met Asn1 5 10
15257PRTMus musculusmisc_feature(1)..(7)0302 light chain CDR2 25Thr
Ala Ser Asn Leu Glu Ser1 5269PRTMus
musculusmisc_feature(1)..(9)0302 light chain CDR3 26Gln Gln Ser Lys
Glu Leu Pro Trp Thr1 52710PRTMus musculusmisc_feature(1)..(10)0311
heavy chain CDR1 27Gly Tyr Ile Phe Thr Asp Tyr Asn Met His1 5
102817PRTMus musculusmisc_feature(1)..(17)0311 heavy chain CDR2
28Glu Ile Asn Pro Asn Asn Gly Val Val Val Tyr Asn Gln Lys Phe Lys1
5 10 15Gly2913PRTMus musculusmisc_feature(1)..(13)0311 heavy chain
CDR3 29Ala Leu Tyr His Ser Asn Phe Gly Trp Tyr Phe Asp Ser1 5
103015PRTMus musculusmisc_feature(1)..(15)0311 light chain CDR1
30Lys Ala Ser Gln Ser Val Asp Tyr Asp Gly Asp Ser His Met Asn1 5 10
15317PRTMus musculusmisc_feature(1)..(7)0311 light chain CDR2 31Thr
Ala Ser Asn Leu Glu Ser1 5329PRTMus
musculusmisc_feature(1)..(9)0311 light chain CDR3 32Gln Gln Gly Asn
Glu Asp Pro Trp Thr1 533449PRTArtificial SequenceSynthetic cAb 0301
heavy chain 33Glu Val Gln Leu Gln Gln Ser Gly Pro Glu Leu Val Arg
Pro Gly Ala1 5 10 15Ser Val Lys Met Ser Cys Lys Ala Ser Gly Tyr Thr
Phe Thr Asp Asn 20 25 30Tyr Met Ile Trp Val Lys Gln Ser His Gly Lys
Ser Leu Glu Trp Ile 35 40 45Gly Asp Ile Asn Pro Tyr Asn Gly Gly Thr
Thr Phe Asn Gln Lys Phe 50 55 60Lys Gly Lys Ala Thr Leu Thr Val Glu
Lys Ser Ser Ser Thr Ala Tyr65 70 75 80Met Gln Leu Asn Ser Leu Thr
Ser Glu Asp Ser Ala Val Tyr Tyr Cys 85 90 95Ala Arg Glu Ser Pro Tyr
Phe Ser Asn Leu Tyr Val Met Asp Tyr Trp 100 105 110Gly Gln Gly Thr
Ser Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro 115 120 125Ser Val
Phe Pro Leu Ala Pro Cys Ser Arg Ser Thr Ser Glu Ser Thr 130 135
140Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val
Thr145 150 155 160Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val
His Thr Phe Pro 165 170 175Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser
Leu Ser Ser Val Val Thr 180 185 190Val Pro Ser Ser Ser Leu Gly Thr
Lys Thr Tyr Thr Cys Asn Val Asp 195 200 205His Lys Pro Ser Asn Thr
Lys Val Asp Lys Arg Val Glu Ser Lys Tyr 210 215 220Gly Pro Pro Cys
Pro Pro Cys Pro Ala Pro Glu Phe Leu Gly Gly Pro225 230 235 240Ser
Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser 245 250
255Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser Gln Glu Asp
260 265 270Pro Glu Val Gln Phe Asn Trp Tyr Val Asp Gly Val Glu Val
His Asn 275 280 285Ala Lys Thr Lys Pro Arg Glu Glu Gln Phe Asn Ser
Thr Tyr Arg Val 290 295 300Val Ser Val Leu Thr Val Leu His Gln Asp
Trp Leu Asn Gly Lys Glu305 310 315 320Tyr Lys Cys Lys Val Ser Asn
Lys Gly Leu Pro Ser Ser Ile Glu Lys 325 330 335Thr Ile Ser Lys Ala
Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr 340 345 350Leu Pro Pro
Ser Gln Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr 355 360 365Cys
Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu 370 375
380Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val
Leu385 390 395 400Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Arg Leu
Thr Val Asp Lys 405 410 415Ser Arg Trp Gln Glu Gly Asn Val Phe Ser
Cys Ser Val Met His Glu 420 425 430Ala Leu His Asn His Tyr Thr Gln
Lys Ser Leu Ser Leu Ser Leu Gly 435 440 445Lys34218PRTArtificial
SequenceSynthetic cAb 0301 light chain 34Asn Ile Val Leu Thr Gln
Ser Pro Ala Ser Leu Ala Val Ser Leu Gly1 5 10 15Gln Arg Ala Thr Ile
Ser Cys Lys Ala Ser Gln Ser Val Asp Tyr Asp 20 25 30Gly Asp Asn Tyr
Met Asn Trp Tyr Gln Gln Lys Pro Gly Gln Pro Pro 35 40 45Lys Leu Leu
Ile Tyr Ala Ala Ser Asn Leu Glu Ser Gly Ile Pro Ala 50 55 60Arg Phe
Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Asn Ile His65 70 75
80Pro Val Glu Glu Glu Asp Ala Ala Thr Tyr Tyr Cys His Leu Ser Asn
85 90 95Glu Asp Leu Ser Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys
Arg 100 105 110Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser
Asp Glu Gln 115 120 125Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu
Leu Asn Asn Phe Tyr 130 135 140Pro Arg Glu Ala Lys Val Gln Trp Lys
Val Asp Asn Ala Leu Gln Ser145 150 155 160Gly Asn Ser Gln Glu Ser
Val Thr Glu Gln Asp Ser Lys Asp Ser Thr 165 170 175Tyr Ser Leu Ser
Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys 180 185 190His Lys
Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro 195 200
205Val Thr Lys Ser Phe Asn Arg Gly Glu Cys 210
21535448PRTArtificial SequenceSynthetic cAb 0302 heavy chain 35Glu
Ile Gln Leu Gln Gln Ser Gly Pro Glu Leu Val Lys Pro Gly Ala1 5 10
15Ser Val Lys Met Ser Cys Lys Ala Ser Gly Tyr Thr Phe Ser Asp Phe
20 25 30Asn Ile His Trp Val Lys Gln Lys Pro Gly Gln Gly Leu Glu Trp
Ile 35 40 45Gly Tyr Ile Asn Pro Tyr Thr Asp Val Thr Val Tyr Asn Glu
Lys Phe 50 55 60Lys Gly Lys Ala Thr Leu Thr Ser Asp Arg Ser Ser Ser
Thr Ala Tyr65 70 75 80Met Asp Leu Ser Ser Leu Thr Ser Glu Asp Ser
Ala Val Tyr Tyr Cys 85 90 95Ala Ser Tyr Phe Asp Gly Thr Phe Asp Tyr
Ala Leu Asp Tyr Trp Gly 100 105 110Gln Gly Thr Ser Ile Thr Val Ser
Ser Ala Ser Thr Lys Gly Pro Ser 115 120 125Val Phe Pro Leu Ala Pro
Cys Ser Arg Ser Thr Ser Glu Ser Thr Ala 130 135 140Ala Leu Gly Cys
Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val145 150 155 160Ser
Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala 165 170
175Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val
180 185 190Pro Ser Ser Ser Leu Gly Thr Lys Thr Tyr Thr Cys Asn Val
Asp His 195 200 205Lys Pro Ser Asn Thr Lys Val Asp Lys Arg Val Glu
Ser Lys Tyr Gly 210 215 220Pro Pro Cys Pro Pro Cys Pro Ala Pro Glu
Phe Leu Gly Gly Pro Ser225 230 235 240Val Phe Leu Phe Pro Pro Lys
Pro Lys Asp Thr Leu Met Ile Ser Arg 245 250 255Thr Pro Glu Val Thr
Cys Val Val Val Asp Val Ser Gln Glu Asp Pro 260 265 270Glu Val Gln
Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala 275 280 285Lys
Thr Lys Pro Arg Glu Glu Gln Phe Asn Ser Thr Tyr Arg Val Val 290 295
300Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu
Tyr305 310 315 320Lys Cys Lys Val Ser Asn Lys Gly Leu Pro Ser Ser
Ile Glu Lys Thr 325 330 335Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu
Pro Gln Val Tyr Thr Leu 340 345 350Pro Pro Ser Gln Glu Glu Met Thr
Lys Asn Gln Val Ser Leu Thr Cys 355 360 365Leu Val Lys Gly Phe Tyr
Pro Ser Asp Ile Ala Val Glu Trp Glu Ser 370 375 380Asn Gly Gln Pro
Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp385 390 395 400Ser
Asp Gly Ser Phe Phe Leu Tyr Ser Arg Leu Thr Val Asp Lys Ser 405 410
415Arg Trp Gln Glu Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala
420 425 430Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Leu
Gly Lys 435 440 44536218PRTArtificial SequenceSynthetic cAb 0302
light chain 36Asp Val Val Val Thr Gln Thr Pro Ala Ser Leu Ala Val
Ser Leu Gly1 5 10 15Gln Arg Ala Thr Ile Ser Cys Arg Ala Ser Glu Ser
Val Asp Asn Tyr 20 25 30Gly Leu Ser Phe Met Asn Trp Phe Gln Gln Lys
Pro Gly Gln Pro Pro 35 40 45Lys Leu Leu Ile Tyr Thr Ala Ser Asn Leu
Glu Ser Gly Ile Pro Ala 50 55 60Arg Phe Ser Gly Gly Gly Ser Arg Thr
Asp Phe Thr Leu Thr Ile Asp65 70 75 80Pro Val Glu Ala Asp Asp Ala
Ala Thr Tyr Phe Cys Gln Gln Ser Lys 85 90 95Glu Leu Pro Trp Thr Phe
Gly Gly Gly Thr Arg Leu Glu Ile Lys Arg 100 105 110Thr Val Ala Ala
Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln 115 120 125Leu Lys
Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr 130 135
140Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln
Ser145 150 155 160Gly Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser
Lys Asp Ser Thr 165 170 175Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser
Lys Ala Asp Tyr Glu Lys 180 185 190His Lys Val Tyr Ala Cys Glu Val
Thr His Gln Gly Leu Ser Ser Pro 195 200 205Val Thr Lys Ser Phe Asn
Arg Gly Glu Cys 210 21537449PRTArtificial SequenceSynthetic cAb
0311 heavy chain 37Glu Ile Gln Leu Gln Gln Ser Gly Pro Asp Leu Met
Lys Pro Gly Ala1 5 10 15Ser Val Lys Met Ser Cys Lys Ala Ser Gly Tyr
Ile Phe Thr Asp Tyr 20 25 30Asn Met His Trp Val Lys Gln Asn Gln Gly
Lys Ser Leu Glu Trp Met 35 40 45Gly Glu Ile Asn Pro Asn Asn Gly Val
Val Val Tyr Asn Gln Lys Phe 50 55 60Lys Gly Thr Thr Thr Leu Thr Val
Asp Lys Ser Ser Ser Thr Ala Tyr65 70 75 80Met Asp Leu His Ser Leu
Thr Ser Glu Asp Ser Ala Val Tyr Tyr Cys 85 90 95Thr Arg Ala Leu Tyr
His Ser Asn Phe Gly Trp Tyr Phe Asp Ser Trp 100 105 110Gly Lys Gly
Thr Thr Leu Thr Val Ser Ser Ala Ser Thr Lys Gly Pro 115 120 125Ser
Val Phe Pro Leu Ala Pro Cys Ser Arg Ser Thr Ser Glu Ser Thr 130 135
140Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val
Thr145 150 155 160Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val
His Thr Phe Pro 165 170 175Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser
Leu Ser Ser Val Val Thr 180 185 190Val Pro Ser Ser Ser Leu Gly Thr
Lys Thr Tyr Thr Cys Asn Val Asp 195 200 205His Lys Pro Ser Asn Thr
Lys Val Asp Lys Arg Val Glu Ser Lys Tyr 210 215 220Gly Pro Pro Cys
Pro Pro Cys Pro Ala Pro Glu Phe Leu Gly Gly Pro225 230 235 240Ser
Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser 245 250
255Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser Gln Glu Asp
260 265 270Pro Glu Val Gln Phe Asn Trp Tyr Val Asp Gly Val Glu Val
His Asn 275 280 285Ala Lys Thr Lys Pro Arg Glu Glu Gln Phe Asn Ser
Thr Tyr Arg Val 290 295 300Val Ser Val Leu Thr Val Leu His Gln Asp
Trp Leu Asn Gly Lys Glu305 310 315 320Tyr Lys Cys Lys Val Ser Asn
Lys Gly Leu Pro Ser Ser Ile Glu Lys 325 330 335Thr Ile Ser Lys Ala
Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr 340 345 350Leu Pro Pro
Ser Gln Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr 355 360 365Cys
Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu 370 375
380Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val
Leu385 390
395 400Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Arg Leu Thr Val Asp
Lys 405 410 415Ser Arg Trp Gln Glu Gly Asn Val Phe Ser Cys Ser Val
Met His Glu 420 425 430Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu
Ser Leu Ser Leu Gly 435 440 445Lys38218PRTArtificial
SequenceSynthetic cAb 0311 light chain 38Asp Ile Val Leu Thr Gln
Ser Pro Ala Ser Leu Ala Val Ser Leu Gly1 5 10 15Gln Arg Ala Thr Ile
Ser Cys Lys Ala Ser Gln Ser Val Asp Tyr Asp 20 25 30Gly Asp Ser His
Met Asn Trp Tyr Gln Gln Lys Pro Gly Gln Pro Pro 35 40 45Lys Leu Leu
Ile Tyr Thr Ala Ser Asn Leu Glu Ser Gly Ile Pro Ala 50 55 60Arg Phe
Ser Gly Ser Gly Ser Gly Ala Asp Phe Thr Leu Thr Ile His65 70 75
80Pro Val Glu Glu Glu Asp Ala Ala Thr Tyr Tyr Cys Gln Gln Gly Asn
85 90 95Glu Asp Pro Trp Thr Phe Gly Gly Gly Thr Arg Leu Glu Ile Lys
Arg 100 105 110Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser
Asp Glu Gln 115 120 125Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu
Leu Asn Asn Phe Tyr 130 135 140Pro Arg Glu Ala Lys Val Gln Trp Lys
Val Asp Asn Ala Leu Gln Ser145 150 155 160Gly Asn Ser Gln Glu Ser
Val Thr Glu Gln Asp Ser Lys Asp Ser Thr 165 170 175Tyr Ser Leu Ser
Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys 180 185 190His Lys
Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro 195 200
205Val Thr Lys Ser Phe Asn Arg Gly Glu Cys 210
21539122PRTArtificial SequenceSynthetic h0301-H0 heavy chain
variable region 39Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys
Lys Pro Gly Ser1 5 10 15Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr
Thr Phe Thr Asp Asn 20 25 30Tyr Met Ile Trp Val Arg Gln Ala Pro Gly
Gln Gly Leu Glu Trp Met 35 40 45Gly Asp Ile Asn Pro Tyr Asn Gly Gly
Thr Thr Phe Asn Gln Lys Phe 50 55 60Lys Gly Arg Val Thr Ile Thr Ala
Asp Lys Ser Thr Ser Thr Ala Tyr65 70 75 80Met Glu Leu Ser Ser Leu
Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95Ala Arg Glu Ser Pro
Tyr Phe Ser Asn Leu Tyr Val Met Asp Tyr Trp 100 105 110Gly Gln Gly
Thr Leu Val Thr Val Ser Ser 115 12040122PRTArtificial
SequenceSynthetic h0301-H1 heavy chain variable region 40Gln Val
Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ser1 5 10 15Ser
Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asp Asn 20 25
30Tyr Met Ile Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met
35 40 45Gly Asp Ile Asn Pro Tyr Asn Gly Gly Thr Thr Phe Asn Gln Lys
Phe 50 55 60Lys Gly Arg Val Thr Ile Thr Val Asp Lys Ser Thr Ser Thr
Ala Tyr65 70 75 80Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala
Val Tyr Tyr Cys 85 90 95Ala Arg Glu Ser Pro Tyr Phe Ser Asn Leu Tyr
Val Met Asp Tyr Trp 100 105 110Gly Gln Gly Thr Leu Val Thr Val Ser
Ser 115 12041122PRTArtificial SequenceSynthetic h0301-H2 heavy
chain variable region 41Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val
Lys Lys Pro Gly Ser1 5 10 15Ser Val Lys Val Ser Cys Lys Ala Ser Gly
Tyr Thr Phe Thr Asp Asn 20 25 30Tyr Met Ile Trp Val Arg Gln Ala Pro
Gly Gln Gly Leu Glu Trp Ile 35 40 45Gly Asp Ile Asn Pro Tyr Asn Gly
Gly Thr Thr Phe Asn Gln Lys Phe 50 55 60Lys Gly Arg Ala Thr Leu Thr
Val Asp Lys Ser Thr Ser Thr Ala Tyr65 70 75 80Met Glu Leu Ser Ser
Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95Ala Arg Glu Ser
Pro Tyr Phe Ser Asn Leu Tyr Val Met Asp Tyr Trp 100 105 110Gly Gln
Gly Thr Leu Val Thr Val Ser Ser 115 12042121PRTArtificial
SequenceSynthetic H0302-H1 heavy chain variable region 42Gln Val
Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ser1 5 10 15Ser
Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Ser Asp Phe 20 25
30Asn Ile His Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met
35 40 45Gly Tyr Ile Asn Pro Tyr Thr Asp Val Thr Val Tyr Asn Glu Lys
Phe 50 55 60Lys Gly Arg Val Thr Ile Thr Ser Asp Lys Ser Thr Ser Thr
Ala Tyr65 70 75 80Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala
Val Tyr Tyr Cys 85 90 95Ala Ser Tyr Phe Asp Gly Thr Phe Asp Tyr Ala
Leu Asp Tyr Trp Gly 100 105 110Gln Gly Thr Leu Val Thr Val Ser Ser
115 12043121PRTArtificial SequenceSynthetic H0302-H2 heavy chain
variable region 43Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys
Lys Pro Gly Ser1 5 10 15Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr
Thr Phe Ser Asp Phe 20 25 30Asn Ile His Trp Val Arg Gln Ala Pro Gly
Gln Gly Leu Glu Trp Ile 35 40 45Gly Tyr Ile Asn Pro Tyr Thr Asp Val
Thr Val Tyr Asn Glu Lys Phe 50 55 60Lys Gly Arg Ala Thr Leu Thr Ser
Asp Lys Ser Thr Ser Thr Ala Tyr65 70 75 80Met Glu Leu Ser Ser Leu
Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95Ala Ser Tyr Phe Asp
Gly Thr Phe Asp Tyr Ala Leu Asp Tyr Trp Gly 100 105 110Gln Gly Thr
Leu Val Thr Val Ser Ser 115 12044122PRTArtificial SequenceSynthetic
H0311-H1 heavy chain variable region 44Gln Val Gln Leu Val Gln Ser
Gly Ala Glu Val Lys Lys Pro Gly Ser1 5 10 15Ser Val Lys Val Ser Cys
Lys Ala Ser Gly Tyr Ile Phe Thr Asp Tyr 20 25 30Asn Met His Trp Val
Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45Gly Glu Ile Asn
Pro Asn Asn Gly Val Val Val Tyr Asn Gln Lys Phe 50 55 60Lys Gly Arg
Val Thr Ile Thr Val Asp Lys Ser Thr Ser Thr Ala Tyr65 70 75 80Met
Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90
95Thr Arg Ala Leu Tyr His Ser Asn Phe Gly Trp Tyr Phe Asp Ser Trp
100 105 110Gly Gln Gly Thr Leu Val Thr Val Ser Ser 115
12045122PRTArtificial SequenceSynthetic H0311-H2 heavy chain
variable region 45Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys
Lys Pro Gly Ser1 5 10 15Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr
Ile Phe Thr Asp Tyr 20 25 30Asn Met His Trp Val Arg Gln Ala Pro Gly
Gln Gly Leu Glu Trp Met 35 40 45Gly Glu Ile Asn Pro Asn Asn Gly Val
Val Val Tyr Asn Gln Lys Phe 50 55 60Lys Gly Thr Thr Thr Leu Thr Val
Asp Lys Ser Thr Ser Thr Ala Tyr65 70 75 80Met Glu Leu Ser Ser Leu
Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95Thr Arg Ala Leu Tyr
His Ser Asn Phe Gly Trp Tyr Phe Asp Ser Trp 100 105 110Gly Gln Gly
Thr Leu Val Thr Val Ser Ser 115 12046111PRTArtificial
SequenceSynthetic h0301-L0 light chain variable region 46Glu Ile
Val Leu Thr Gln Ser Pro Ala Thr Leu Ser Leu Ser Pro Gly1 5 10 15Glu
Arg Ala Thr Leu Ser Cys Lys Ala Ser Gln Ser Val Asp Tyr Asp 20 25
30Gly Asp Asn Tyr Met Asn Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro
35 40 45Arg Leu Leu Ile Tyr Ala Ala Ser Asn Leu Glu Ser Gly Ile Pro
Ala 50 55 60Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr
Ile Ser65 70 75 80Ser Leu Glu Pro Glu Asp Phe Ala Val Tyr Tyr Cys
His Leu Ser Asn 85 90 95Glu Asp Leu Ser Thr Phe Gly Gly Gly Thr Lys
Val Glu Ile Lys 100 105 11047111PRTArtificial SequenceSynthetic
h0301-L1 light chain variable region 47Asn Ile Val Leu Thr Gln Ser
Pro Ala Thr Leu Ser Leu Ser Pro Gly1 5 10 15Glu Arg Ala Thr Leu Ser
Cys Lys Ala Ser Gln Ser Val Asp Tyr Asp 20 25 30Gly Asp Asn Tyr Met
Asn Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro 35 40 45Arg Leu Leu Ile
Tyr Ala Ala Ser Asn Leu Glu Ser Gly Ile Pro Ala 50 55 60Arg Phe Ser
Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser65 70 75 80Ser
Leu Glu Pro Glu Asp Phe Ala Val Tyr Tyr Cys His Leu Ser Asn 85 90
95Glu Asp Leu Ser Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys 100
105 11048111PRTArtificial SequenceSynthetic H0302-L0 light chain
variable region 48Glu Ile Val Leu Thr Gln Ser Pro Ala Thr Leu Ser
Leu Ser Pro Gly1 5 10 15Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Glu
Ser Val Asp Asn Tyr 20 25 30Gly Leu Ser Phe Met Asn Trp Tyr Gln Gln
Lys Pro Gly Gln Ala Pro 35 40 45Arg Leu Leu Ile Tyr Thr Ala Ser Asn
Leu Glu Ser Gly Ile Pro Ala 50 55 60Arg Phe Ser Gly Ser Gly Ser Gly
Thr Asp Phe Thr Leu Thr Ile Ser65 70 75 80Ser Leu Glu Pro Glu Asp
Phe Ala Val Tyr Tyr Cys Gln Gln Ser Lys 85 90 95Glu Leu Pro Trp Thr
Phe Gly Gln Gly Thr Lys Val Glu Ile Lys 100 105
11049111PRTArtificial SequenceSynthetic H0302-L1 light chain
variable region 49Glu Ile Val Leu Thr Gln Ser Pro Ala Thr Leu Ser
Leu Ser Pro Gly1 5 10 15Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Glu
Ser Val Asp Asn Tyr 20 25 30Gly Leu Ser Phe Met Asn Trp Tyr Gln Gln
Lys Pro Gly Gln Ala Pro 35 40 45Arg Leu Leu Ile Tyr Thr Ala Ser Asn
Leu Glu Ser Gly Ile Pro Ala 50 55 60Arg Phe Ser Gly Ser Gly Ser Arg
Thr Asp Phe Thr Leu Thr Ile Ser65 70 75 80Ser Leu Glu Pro Glu Asp
Phe Ala Val Tyr Tyr Cys Gln Gln Ser Lys 85 90 95Glu Leu Pro Trp Thr
Phe Gly Gln Gly Thr Lys Val Glu Ile Lys 100 105
11050111PRTArtificial SequenceSynthetic H0302-L2 light chain
variable region 50Glu Ile Val Val Thr Gln Ser Pro Ala Thr Leu Ser
Leu Ser Pro Gly1 5 10 15Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Glu
Ser Val Asp Asn Tyr 20 25 30Gly Leu Ser Phe Met Asn Trp Phe Gln Gln
Lys Pro Gly Gln Ala Pro 35 40 45Arg Leu Leu Ile Tyr Thr Ala Ser Asn
Leu Glu Ser Gly Ile Pro Ala 50 55 60Arg Phe Ser Gly Ser Gly Ser Arg
Thr Asp Phe Thr Leu Thr Ile Ser65 70 75 80Ser Leu Glu Pro Glu Asp
Phe Ala Val Tyr Tyr Cys Gln Gln Ser Lys 85 90 95Glu Leu Pro Trp Thr
Phe Gly Gln Gly Thr Lys Val Glu Ile Lys 100 105
11051111PRTArtificial SequenceSynthetic H0311-L0 light chain
variable region 51Glu Ile Val Leu Thr Gln Ser Pro Ala Thr Leu Ser
Leu Ser Pro Gly1 5 10 15Glu Arg Ala Thr Leu Ser Cys Lys Ala Ser Gln
Ser Val Asp Tyr Asp 20 25 30Gly Asp Ser His Met Asn Trp Tyr Gln Gln
Lys Pro Gly Gln Ala Pro 35 40 45Arg Leu Leu Ile Tyr Thr Ala Ser Asn
Leu Glu Ser Gly Ile Pro Ala 50 55 60Arg Phe Ser Gly Ser Gly Ser Gly
Thr Asp Phe Thr Leu Thr Ile Ser65 70 75 80Ser Leu Glu Pro Glu Asp
Phe Ala Val Tyr Tyr Cys Gln Gln Gly Asn 85 90 95Glu Asp Pro Trp Thr
Phe Gly Gln Gly Thr Lys Val Glu Ile Lys 100 105
11052111PRTArtificial SequenceSynthetic H0311-L1 light chain
variable region 52Asp Ile Val Leu Thr Gln Ser Pro Ala Thr Leu Ser
Leu Ser Pro Gly1 5 10 15Glu Arg Ala Thr Leu Ser Cys Lys Ala Ser Gln
Ser Val Asp Tyr Asp 20 25 30Gly Asp Ser His Met Asn Trp Tyr Gln Gln
Lys Pro Gly Gln Ala Pro 35 40 45Arg Leu Leu Ile Tyr Thr Ala Ser Asn
Leu Glu Ser Gly Ile Pro Ala 50 55 60Arg Phe Ser Gly Ser Gly Ser Gly
Ala Asp Phe Thr Leu Thr Ile Ser65 70 75 80Ser Leu Glu Pro Glu Asp
Phe Ala Val Tyr Tyr Cys Gln Gln Gly Asn 85 90 95Glu Asp Pro Trp Thr
Phe Gly Gln Gly Thr Lys Val Glu Ile Lys 100 105
11053449PRTArtificial SequenceSynthetic h0301-H0 heavy chain 53Gln
Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ser1 5 10
15Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asp Asn
20 25 30Tyr Met Ile Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp
Met 35 40 45Gly Asp Ile Asn Pro Tyr Asn Gly Gly Thr Thr Phe Asn Gln
Lys Phe 50 55 60Lys Gly Arg Val Thr Ile Thr Ala Asp Lys Ser Thr Ser
Thr Ala Tyr65 70 75 80Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr
Ala Val Tyr Tyr Cys 85 90 95Ala Arg Glu Ser Pro Tyr Phe Ser Asn Leu
Tyr Val Met Asp Tyr Trp 100 105 110Gly Gln Gly Thr Leu Val Thr Val
Ser Ser Ala Ser Thr Lys Gly Pro 115 120 125Ser Val Phe Pro Leu Ala
Pro Cys Ser Arg Ser Thr Ser Glu Ser Thr 130 135 140Ala Ala Leu Gly
Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr145 150 155 160Val
Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro 165 170
175Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr
180 185 190Val Pro Ser Ser Ser Leu Gly Thr Lys Thr Tyr Thr Cys Asn
Val Asp 195 200 205His Lys Pro Ser Asn Thr Lys Val Asp Lys Arg Val
Glu Ser Lys Tyr 210 215 220Gly Pro Pro Cys Pro Pro Cys Pro Ala Pro
Glu Phe Leu Gly Gly Pro225 230 235 240Ser Val Phe Leu Phe Pro Pro
Lys Pro Lys Asp Thr Leu Met Ile Ser 245 250 255Arg Thr Pro Glu Val
Thr Cys Val Val Val Asp Val Ser Gln Glu Asp 260 265 270Pro Glu Val
Gln Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn 275 280 285Ala
Lys Thr Lys Pro Arg Glu Glu Gln Phe Asn Ser Thr Tyr Arg Val 290 295
300Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys
Glu305 310 315 320Tyr Lys Cys Lys Val Ser Asn Lys Gly Leu Pro Ser
Ser Ile Glu Lys 325 330 335Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg
Glu Pro Gln Val Tyr Thr 340 345 350Leu Pro Pro Ser Gln Glu Glu Met
Thr Lys Asn Gln Val Ser Leu Thr 355 360 365Cys Leu Val Lys Gly Phe
Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu 370 375 380Ser Asn Gly Gln
Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu385 390 395 400Asp
Ser Asp Gly Ser Phe Phe Leu Tyr Ser Arg Leu Thr Val Asp Lys 405
410
415Ser Arg Trp Gln Glu Gly Asn Val Phe Ser Cys Ser Val Met His Glu
420 425 430Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser
Leu Gly 435 440 445Lys54449PRTArtificial SequenceSynthetic h0301-H1
heavy chain 54Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys
Pro Gly Ser1 5 10 15Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr
Phe Thr Asp Asn 20 25 30Tyr Met Ile Trp Val Arg Gln Ala Pro Gly Gln
Gly Leu Glu Trp Met 35 40 45Gly Asp Ile Asn Pro Tyr Asn Gly Gly Thr
Thr Phe Asn Gln Lys Phe 50 55 60Lys Gly Arg Val Thr Ile Thr Val Asp
Lys Ser Thr Ser Thr Ala Tyr65 70 75 80Met Glu Leu Ser Ser Leu Arg
Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95Ala Arg Glu Ser Pro Tyr
Phe Ser Asn Leu Tyr Val Met Asp Tyr Trp 100 105 110Gly Gln Gly Thr
Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro 115 120 125Ser Val
Phe Pro Leu Ala Pro Cys Ser Arg Ser Thr Ser Glu Ser Thr 130 135
140Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val
Thr145 150 155 160Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val
His Thr Phe Pro 165 170 175Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser
Leu Ser Ser Val Val Thr 180 185 190Val Pro Ser Ser Ser Leu Gly Thr
Lys Thr Tyr Thr Cys Asn Val Asp 195 200 205His Lys Pro Ser Asn Thr
Lys Val Asp Lys Arg Val Glu Ser Lys Tyr 210 215 220Gly Pro Pro Cys
Pro Pro Cys Pro Ala Pro Glu Phe Leu Gly Gly Pro225 230 235 240Ser
Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser 245 250
255Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser Gln Glu Asp
260 265 270Pro Glu Val Gln Phe Asn Trp Tyr Val Asp Gly Val Glu Val
His Asn 275 280 285Ala Lys Thr Lys Pro Arg Glu Glu Gln Phe Asn Ser
Thr Tyr Arg Val 290 295 300Val Ser Val Leu Thr Val Leu His Gln Asp
Trp Leu Asn Gly Lys Glu305 310 315 320Tyr Lys Cys Lys Val Ser Asn
Lys Gly Leu Pro Ser Ser Ile Glu Lys 325 330 335Thr Ile Ser Lys Ala
Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr 340 345 350Leu Pro Pro
Ser Gln Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr 355 360 365Cys
Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu 370 375
380Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val
Leu385 390 395 400Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Arg Leu
Thr Val Asp Lys 405 410 415Ser Arg Trp Gln Glu Gly Asn Val Phe Ser
Cys Ser Val Met His Glu 420 425 430Ala Leu His Asn His Tyr Thr Gln
Lys Ser Leu Ser Leu Ser Leu Gly 435 440 445Lys55449PRTArtificial
SequenceSynthetic h0301-H2 heavy chain 55Gln Val Gln Leu Val Gln
Ser Gly Ala Glu Val Lys Lys Pro Gly Ser1 5 10 15Ser Val Lys Val Ser
Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asp Asn 20 25 30Tyr Met Ile Trp
Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Ile 35 40 45Gly Asp Ile
Asn Pro Tyr Asn Gly Gly Thr Thr Phe Asn Gln Lys Phe 50 55 60Lys Gly
Arg Ala Thr Leu Thr Val Asp Lys Ser Thr Ser Thr Ala Tyr65 70 75
80Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95Ala Arg Glu Ser Pro Tyr Phe Ser Asn Leu Tyr Val Met Asp Tyr
Trp 100 105 110Gly Gln Gly Thr Leu Val Thr Val Ser Ser Ala Ser Thr
Lys Gly Pro 115 120 125Ser Val Phe Pro Leu Ala Pro Cys Ser Arg Ser
Thr Ser Glu Ser Thr 130 135 140Ala Ala Leu Gly Cys Leu Val Lys Asp
Tyr Phe Pro Glu Pro Val Thr145 150 155 160Val Ser Trp Asn Ser Gly
Ala Leu Thr Ser Gly Val His Thr Phe Pro 165 170 175Ala Val Leu Gln
Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr 180 185 190Val Pro
Ser Ser Ser Leu Gly Thr Lys Thr Tyr Thr Cys Asn Val Asp 195 200
205His Lys Pro Ser Asn Thr Lys Val Asp Lys Arg Val Glu Ser Lys Tyr
210 215 220Gly Pro Pro Cys Pro Pro Cys Pro Ala Pro Glu Phe Leu Gly
Gly Pro225 230 235 240Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp
Thr Leu Met Ile Ser 245 250 255Arg Thr Pro Glu Val Thr Cys Val Val
Val Asp Val Ser Gln Glu Asp 260 265 270Pro Glu Val Gln Phe Asn Trp
Tyr Val Asp Gly Val Glu Val His Asn 275 280 285Ala Lys Thr Lys Pro
Arg Glu Glu Gln Phe Asn Ser Thr Tyr Arg Val 290 295 300Val Ser Val
Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu305 310 315
320Tyr Lys Cys Lys Val Ser Asn Lys Gly Leu Pro Ser Ser Ile Glu Lys
325 330 335Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val
Tyr Thr 340 345 350Leu Pro Pro Ser Gln Glu Glu Met Thr Lys Asn Gln
Val Ser Leu Thr 355 360 365Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp
Ile Ala Val Glu Trp Glu 370 375 380Ser Asn Gly Gln Pro Glu Asn Asn
Tyr Lys Thr Thr Pro Pro Val Leu385 390 395 400Asp Ser Asp Gly Ser
Phe Phe Leu Tyr Ser Arg Leu Thr Val Asp Lys 405 410 415Ser Arg Trp
Gln Glu Gly Asn Val Phe Ser Cys Ser Val Met His Glu 420 425 430Ala
Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Leu Gly 435 440
445Lys56448PRTArtificial SequenceSynthetic H0302-H1 heavy chain
56Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ser1
5 10 15Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Ser Asp
Phe 20 25 30Asn Ile His Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu
Trp Met 35 40 45Gly Tyr Ile Asn Pro Tyr Thr Asp Val Thr Val Tyr Asn
Glu Lys Phe 50 55 60Lys Gly Arg Val Thr Ile Thr Ser Asp Lys Ser Thr
Ser Thr Ala Tyr65 70 75 80Met Glu Leu Ser Ser Leu Arg Ser Glu Asp
Thr Ala Val Tyr Tyr Cys 85 90 95Ala Ser Tyr Phe Asp Gly Thr Phe Asp
Tyr Ala Leu Asp Tyr Trp Gly 100 105 110Gln Gly Thr Leu Val Thr Val
Ser Ser Ala Ser Thr Lys Gly Pro Ser 115 120 125Val Phe Pro Leu Ala
Pro Cys Ser Arg Ser Thr Ser Glu Ser Thr Ala 130 135 140Ala Leu Gly
Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val145 150 155
160Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala
165 170 175Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val
Thr Val 180 185 190Pro Ser Ser Ser Leu Gly Thr Lys Thr Tyr Thr Cys
Asn Val Asp His 195 200 205Lys Pro Ser Asn Thr Lys Val Asp Lys Arg
Val Glu Ser Lys Tyr Gly 210 215 220Pro Pro Cys Pro Pro Cys Pro Ala
Pro Glu Phe Leu Gly Gly Pro Ser225 230 235 240Val Phe Leu Phe Pro
Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg 245 250 255Thr Pro Glu
Val Thr Cys Val Val Val Asp Val Ser Gln Glu Asp Pro 260 265 270Glu
Val Gln Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala 275 280
285Lys Thr Lys Pro Arg Glu Glu Gln Phe Asn Ser Thr Tyr Arg Val Val
290 295 300Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys
Glu Tyr305 310 315 320Lys Cys Lys Val Ser Asn Lys Gly Leu Pro Ser
Ser Ile Glu Lys Thr 325 330 335Ile Ser Lys Ala Lys Gly Gln Pro Arg
Glu Pro Gln Val Tyr Thr Leu 340 345 350Pro Pro Ser Gln Glu Glu Met
Thr Lys Asn Gln Val Ser Leu Thr Cys 355 360 365Leu Val Lys Gly Phe
Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser 370 375 380Asn Gly Gln
Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp385 390 395
400Ser Asp Gly Ser Phe Phe Leu Tyr Ser Arg Leu Thr Val Asp Lys Ser
405 410 415Arg Trp Gln Glu Gly Asn Val Phe Ser Cys Ser Val Met His
Glu Ala 420 425 430Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu
Ser Leu Gly Lys 435 440 44557448PRTArtificial SequenceSynthetic
H0302-H2 heavy chain 57Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val
Lys Lys Pro Gly Ser1 5 10 15Ser Val Lys Val Ser Cys Lys Ala Ser Gly
Tyr Thr Phe Ser Asp Phe 20 25 30Asn Ile His Trp Val Arg Gln Ala Pro
Gly Gln Gly Leu Glu Trp Ile 35 40 45Gly Tyr Ile Asn Pro Tyr Thr Asp
Val Thr Val Tyr Asn Glu Lys Phe 50 55 60Lys Gly Arg Ala Thr Leu Thr
Ser Asp Lys Ser Thr Ser Thr Ala Tyr65 70 75 80Met Glu Leu Ser Ser
Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95Ala Ser Tyr Phe
Asp Gly Thr Phe Asp Tyr Ala Leu Asp Tyr Trp Gly 100 105 110Gln Gly
Thr Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser 115 120
125Val Phe Pro Leu Ala Pro Cys Ser Arg Ser Thr Ser Glu Ser Thr Ala
130 135 140Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val
Thr Val145 150 155 160Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val
His Thr Phe Pro Ala 165 170 175Val Leu Gln Ser Ser Gly Leu Tyr Ser
Leu Ser Ser Val Val Thr Val 180 185 190Pro Ser Ser Ser Leu Gly Thr
Lys Thr Tyr Thr Cys Asn Val Asp His 195 200 205Lys Pro Ser Asn Thr
Lys Val Asp Lys Arg Val Glu Ser Lys Tyr Gly 210 215 220Pro Pro Cys
Pro Pro Cys Pro Ala Pro Glu Phe Leu Gly Gly Pro Ser225 230 235
240Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg
245 250 255Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser Gln Glu
Asp Pro 260 265 270Glu Val Gln Phe Asn Trp Tyr Val Asp Gly Val Glu
Val His Asn Ala 275 280 285Lys Thr Lys Pro Arg Glu Glu Gln Phe Asn
Ser Thr Tyr Arg Val Val 290 295 300Ser Val Leu Thr Val Leu His Gln
Asp Trp Leu Asn Gly Lys Glu Tyr305 310 315 320Lys Cys Lys Val Ser
Asn Lys Gly Leu Pro Ser Ser Ile Glu Lys Thr 325 330 335Ile Ser Lys
Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu 340 345 350Pro
Pro Ser Gln Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr Cys 355 360
365Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser
370 375 380Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val
Leu Asp385 390 395 400Ser Asp Gly Ser Phe Phe Leu Tyr Ser Arg Leu
Thr Val Asp Lys Ser 405 410 415Arg Trp Gln Glu Gly Asn Val Phe Ser
Cys Ser Val Met His Glu Ala 420 425 430Leu His Asn His Tyr Thr Gln
Lys Ser Leu Ser Leu Ser Leu Gly Lys 435 440 44558449PRTArtificial
SequenceSynthetic H0311-H1 heavy chain 58Gln Val Gln Leu Val Gln
Ser Gly Ala Glu Val Lys Lys Pro Gly Ser1 5 10 15Ser Val Lys Val Ser
Cys Lys Ala Ser Gly Tyr Ile Phe Thr Asp Tyr 20 25 30Asn Met His Trp
Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45Gly Glu Ile
Asn Pro Asn Asn Gly Val Val Val Tyr Asn Gln Lys Phe 50 55 60Lys Gly
Arg Val Thr Ile Thr Val Asp Lys Ser Thr Ser Thr Ala Tyr65 70 75
80Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95Thr Arg Ala Leu Tyr His Ser Asn Phe Gly Trp Tyr Phe Asp Ser
Trp 100 105 110Gly Gln Gly Thr Leu Val Thr Val Ser Ser Ala Ser Thr
Lys Gly Pro 115 120 125Ser Val Phe Pro Leu Ala Pro Cys Ser Arg Ser
Thr Ser Glu Ser Thr 130 135 140Ala Ala Leu Gly Cys Leu Val Lys Asp
Tyr Phe Pro Glu Pro Val Thr145 150 155 160Val Ser Trp Asn Ser Gly
Ala Leu Thr Ser Gly Val His Thr Phe Pro 165 170 175Ala Val Leu Gln
Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr 180 185 190Val Pro
Ser Ser Ser Leu Gly Thr Lys Thr Tyr Thr Cys Asn Val Asp 195 200
205His Lys Pro Ser Asn Thr Lys Val Asp Lys Arg Val Glu Ser Lys Tyr
210 215 220Gly Pro Pro Cys Pro Pro Cys Pro Ala Pro Glu Phe Leu Gly
Gly Pro225 230 235 240Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp
Thr Leu Met Ile Ser 245 250 255Arg Thr Pro Glu Val Thr Cys Val Val
Val Asp Val Ser Gln Glu Asp 260 265 270Pro Glu Val Gln Phe Asn Trp
Tyr Val Asp Gly Val Glu Val His Asn 275 280 285Ala Lys Thr Lys Pro
Arg Glu Glu Gln Phe Asn Ser Thr Tyr Arg Val 290 295 300Val Ser Val
Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu305 310 315
320Tyr Lys Cys Lys Val Ser Asn Lys Gly Leu Pro Ser Ser Ile Glu Lys
325 330 335Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val
Tyr Thr 340 345 350Leu Pro Pro Ser Gln Glu Glu Met Thr Lys Asn Gln
Val Ser Leu Thr 355 360 365Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp
Ile Ala Val Glu Trp Glu 370 375 380Ser Asn Gly Gln Pro Glu Asn Asn
Tyr Lys Thr Thr Pro Pro Val Leu385 390 395 400Asp Ser Asp Gly Ser
Phe Phe Leu Tyr Ser Arg Leu Thr Val Asp Lys 405 410 415Ser Arg Trp
Gln Glu Gly Asn Val Phe Ser Cys Ser Val Met His Glu 420 425 430Ala
Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Leu Gly 435 440
445Lys59449PRTArtificial SequenceSynthetic H0311-H2 heavy chain
59Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ser1
5 10 15Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Ile Phe Thr Asp
Tyr 20 25 30Asn Met His Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu
Trp Met 35 40 45Gly Glu Ile Asn Pro Asn Asn Gly Val Val Val Tyr Asn
Gln Lys Phe 50 55 60Lys Gly Thr Thr Thr Leu Thr Val Asp Lys Ser Thr
Ser Thr Ala Tyr65 70 75 80Met Glu Leu Ser Ser Leu Arg Ser Glu Asp
Thr Ala Val Tyr Tyr Cys 85 90 95Thr Arg Ala Leu Tyr His Ser Asn Phe
Gly Trp Tyr Phe Asp Ser Trp 100 105 110Gly Gln Gly Thr Leu Val Thr
Val Ser Ser Ala Ser Thr Lys Gly Pro 115 120 125Ser Val Phe Pro Leu
Ala Pro Cys Ser Arg Ser Thr Ser Glu Ser Thr 130 135 140Ala Ala Leu
Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr145
150 155 160Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr
Phe Pro 165 170 175Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser
Ser Val Val Thr 180 185 190Val Pro Ser Ser Ser Leu Gly Thr Lys Thr
Tyr Thr Cys Asn Val Asp 195 200 205His Lys Pro Ser Asn Thr Lys Val
Asp Lys Arg Val Glu Ser Lys Tyr 210 215 220Gly Pro Pro Cys Pro Pro
Cys Pro Ala Pro Glu Phe Leu Gly Gly Pro225 230 235 240Ser Val Phe
Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser 245 250 255Arg
Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser Gln Glu Asp 260 265
270Pro Glu Val Gln Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn
275 280 285Ala Lys Thr Lys Pro Arg Glu Glu Gln Phe Asn Ser Thr Tyr
Arg Val 290 295 300Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu
Asn Gly Lys Glu305 310 315 320Tyr Lys Cys Lys Val Ser Asn Lys Gly
Leu Pro Ser Ser Ile Glu Lys 325 330 335Thr Ile Ser Lys Ala Lys Gly
Gln Pro Arg Glu Pro Gln Val Tyr Thr 340 345 350Leu Pro Pro Ser Gln
Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr 355 360 365Cys Leu Val
Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu 370 375 380Ser
Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu385 390
395 400Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Arg Leu Thr Val Asp
Lys 405 410 415Ser Arg Trp Gln Glu Gly Asn Val Phe Ser Cys Ser Val
Met His Glu 420 425 430Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu
Ser Leu Ser Leu Gly 435 440 445Lys60218PRTArtificial
SequenceSynthetic h0301-L0 light chain 60Glu Ile Val Leu Thr Gln
Ser Pro Ala Thr Leu Ser Leu Ser Pro Gly1 5 10 15Glu Arg Ala Thr Leu
Ser Cys Lys Ala Ser Gln Ser Val Asp Tyr Asp 20 25 30Gly Asp Asn Tyr
Met Asn Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro 35 40 45Arg Leu Leu
Ile Tyr Ala Ala Ser Asn Leu Glu Ser Gly Ile Pro Ala 50 55 60Arg Phe
Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser65 70 75
80Ser Leu Glu Pro Glu Asp Phe Ala Val Tyr Tyr Cys His Leu Ser Asn
85 90 95Glu Asp Leu Ser Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys
Arg 100 105 110Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser
Asp Glu Gln 115 120 125Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu
Leu Asn Asn Phe Tyr 130 135 140Pro Arg Glu Ala Lys Val Gln Trp Lys
Val Asp Asn Ala Leu Gln Ser145 150 155 160Gly Asn Ser Gln Glu Ser
Val Thr Glu Gln Asp Ser Lys Asp Ser Thr 165 170 175Tyr Ser Leu Ser
Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys 180 185 190His Lys
Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro 195 200
205Val Thr Lys Ser Phe Asn Arg Gly Glu Cys 210
21561218PRTArtificial SequenceSynthetic h0301-L1 light chain 61Asn
Ile Val Leu Thr Gln Ser Pro Ala Thr Leu Ser Leu Ser Pro Gly1 5 10
15Glu Arg Ala Thr Leu Ser Cys Lys Ala Ser Gln Ser Val Asp Tyr Asp
20 25 30Gly Asp Asn Tyr Met Asn Trp Tyr Gln Gln Lys Pro Gly Gln Ala
Pro 35 40 45Arg Leu Leu Ile Tyr Ala Ala Ser Asn Leu Glu Ser Gly Ile
Pro Ala 50 55 60Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu
Thr Ile Ser65 70 75 80Ser Leu Glu Pro Glu Asp Phe Ala Val Tyr Tyr
Cys His Leu Ser Asn 85 90 95Glu Asp Leu Ser Thr Phe Gly Gly Gly Thr
Lys Val Glu Ile Lys Arg 100 105 110Thr Val Ala Ala Pro Ser Val Phe
Ile Phe Pro Pro Ser Asp Glu Gln 115 120 125Leu Lys Ser Gly Thr Ala
Ser Val Val Cys Leu Leu Asn Asn Phe Tyr 130 135 140Pro Arg Glu Ala
Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser145 150 155 160Gly
Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr 165 170
175Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys
180 185 190His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser
Ser Pro 195 200 205Val Thr Lys Ser Phe Asn Arg Gly Glu Cys 210
21562218PRTArtificial SequenceSynthetic H0302-L0 light chain 62Glu
Ile Val Leu Thr Gln Ser Pro Ala Thr Leu Ser Leu Ser Pro Gly1 5 10
15Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Glu Ser Val Asp Asn Tyr
20 25 30Gly Leu Ser Phe Met Asn Trp Tyr Gln Gln Lys Pro Gly Gln Ala
Pro 35 40 45Arg Leu Leu Ile Tyr Thr Ala Ser Asn Leu Glu Ser Gly Ile
Pro Ala 50 55 60Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu
Thr Ile Ser65 70 75 80Ser Leu Glu Pro Glu Asp Phe Ala Val Tyr Tyr
Cys Gln Gln Ser Lys 85 90 95Glu Leu Pro Trp Thr Phe Gly Gln Gly Thr
Lys Val Glu Ile Lys Arg 100 105 110Thr Val Ala Ala Pro Ser Val Phe
Ile Phe Pro Pro Ser Asp Glu Gln 115 120 125Leu Lys Ser Gly Thr Ala
Ser Val Val Cys Leu Leu Asn Asn Phe Tyr 130 135 140Pro Arg Glu Ala
Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser145 150 155 160Gly
Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr 165 170
175Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys
180 185 190His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser
Ser Pro 195 200 205Val Thr Lys Ser Phe Asn Arg Gly Glu Cys 210
21563218PRTArtificial SequenceSynthetic H0302-L1 light chain 63Glu
Ile Val Leu Thr Gln Ser Pro Ala Thr Leu Ser Leu Ser Pro Gly1 5 10
15Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Glu Ser Val Asp Asn Tyr
20 25 30Gly Leu Ser Phe Met Asn Trp Tyr Gln Gln Lys Pro Gly Gln Ala
Pro 35 40 45Arg Leu Leu Ile Tyr Thr Ala Ser Asn Leu Glu Ser Gly Ile
Pro Ala 50 55 60Arg Phe Ser Gly Ser Gly Ser Arg Thr Asp Phe Thr Leu
Thr Ile Ser65 70 75 80Ser Leu Glu Pro Glu Asp Phe Ala Val Tyr Tyr
Cys Gln Gln Ser Lys 85 90 95Glu Leu Pro Trp Thr Phe Gly Gln Gly Thr
Lys Val Glu Ile Lys Arg 100 105 110Thr Val Ala Ala Pro Ser Val Phe
Ile Phe Pro Pro Ser Asp Glu Gln 115 120 125Leu Lys Ser Gly Thr Ala
Ser Val Val Cys Leu Leu Asn Asn Phe Tyr 130 135 140Pro Arg Glu Ala
Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser145 150 155 160Gly
Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr 165 170
175Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys
180 185 190His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser
Ser Pro 195 200 205Val Thr Lys Ser Phe Asn Arg Gly Glu Cys 210
21564218PRTArtificial SequenceSynthetic H0302-L2 light chain 64Glu
Ile Val Val Thr Gln Ser Pro Ala Thr Leu Ser Leu Ser Pro Gly1 5 10
15Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Glu Ser Val Asp Asn Tyr
20 25 30Gly Leu Ser Phe Met Asn Trp Phe Gln Gln Lys Pro Gly Gln Ala
Pro 35 40 45Arg Leu Leu Ile Tyr Thr Ala Ser Asn Leu Glu Ser Gly Ile
Pro Ala 50 55 60Arg Phe Ser Gly Ser Gly Ser Arg Thr Asp Phe Thr Leu
Thr Ile Ser65 70 75 80Ser Leu Glu Pro Glu Asp Phe Ala Val Tyr Tyr
Cys Gln Gln Ser Lys 85 90 95Glu Leu Pro Trp Thr Phe Gly Gln Gly Thr
Lys Val Glu Ile Lys Arg 100 105 110Thr Val Ala Ala Pro Ser Val Phe
Ile Phe Pro Pro Ser Asp Glu Gln 115 120 125Leu Lys Ser Gly Thr Ala
Ser Val Val Cys Leu Leu Asn Asn Phe Tyr 130 135 140Pro Arg Glu Ala
Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser145 150 155 160Gly
Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr 165 170
175Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys
180 185 190His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser
Ser Pro 195 200 205Val Thr Lys Ser Phe Asn Arg Gly Glu Cys 210
21565218PRTArtificial SequenceSynthetic H0311-L0 light chain 65Glu
Ile Val Leu Thr Gln Ser Pro Ala Thr Leu Ser Leu Ser Pro Gly1 5 10
15Glu Arg Ala Thr Leu Ser Cys Lys Ala Ser Gln Ser Val Asp Tyr Asp
20 25 30Gly Asp Ser His Met Asn Trp Tyr Gln Gln Lys Pro Gly Gln Ala
Pro 35 40 45Arg Leu Leu Ile Tyr Thr Ala Ser Asn Leu Glu Ser Gly Ile
Pro Ala 50 55 60Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu
Thr Ile Ser65 70 75 80Ser Leu Glu Pro Glu Asp Phe Ala Val Tyr Tyr
Cys Gln Gln Gly Asn 85 90 95Glu Asp Pro Trp Thr Phe Gly Gln Gly Thr
Lys Val Glu Ile Lys Arg 100 105 110Thr Val Ala Ala Pro Ser Val Phe
Ile Phe Pro Pro Ser Asp Glu Gln 115 120 125Leu Lys Ser Gly Thr Ala
Ser Val Val Cys Leu Leu Asn Asn Phe Tyr 130 135 140Pro Arg Glu Ala
Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser145 150 155 160Gly
Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr 165 170
175Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys
180 185 190His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser
Ser Pro 195 200 205Val Thr Lys Ser Phe Asn Arg Gly Glu Cys 210
21566218PRTArtificial SequenceSynthetic H0311-L1 light chain 66Asp
Ile Val Leu Thr Gln Ser Pro Ala Thr Leu Ser Leu Ser Pro Gly1 5 10
15Glu Arg Ala Thr Leu Ser Cys Lys Ala Ser Gln Ser Val Asp Tyr Asp
20 25 30Gly Asp Ser His Met Asn Trp Tyr Gln Gln Lys Pro Gly Gln Ala
Pro 35 40 45Arg Leu Leu Ile Tyr Thr Ala Ser Asn Leu Glu Ser Gly Ile
Pro Ala 50 55 60Arg Phe Ser Gly Ser Gly Ser Gly Ala Asp Phe Thr Leu
Thr Ile Ser65 70 75 80Ser Leu Glu Pro Glu Asp Phe Ala Val Tyr Tyr
Cys Gln Gln Gly Asn 85 90 95Glu Asp Pro Trp Thr Phe Gly Gln Gly Thr
Lys Val Glu Ile Lys Arg 100 105 110Thr Val Ala Ala Pro Ser Val Phe
Ile Phe Pro Pro Ser Asp Glu Gln 115 120 125Leu Lys Ser Gly Thr Ala
Ser Val Val Cys Leu Leu Asn Asn Phe Tyr 130 135 140Pro Arg Glu Ala
Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser145 150 155 160Gly
Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr 165 170
175Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys
180 185 190His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser
Ser Pro 195 200 205Val Thr Lys Ser Phe Asn Arg Gly Glu Cys 210
21567158PRTHomo sapiensmisc_feature(1)..(158)Human CSF1 67Glu Glu
Val Ser Glu Tyr Cys Ser His Met Ile Gly Ser Gly His Leu1 5 10 15Gln
Ser Leu Gln Arg Leu Ile Asp Ser Gln Met Glu Thr Ser Cys Gln 20 25
30Ile Thr Phe Glu Phe Val Asp Gln Glu Gln Leu Lys Asp Pro Val Cys
35 40 45Tyr Leu Lys Lys Ala Phe Leu Leu Val Gln Asp Ile Met Glu Asp
Thr 50 55 60Met Arg Phe Arg Asp Asn Thr Pro Asn Ala Ile Ala Ile Val
Gln Leu65 70 75 80Gln Glu Leu Ser Leu Arg Leu Lys Ser Cys Phe Thr
Lys Asp Tyr Glu 85 90 95Glu His Asp Lys Ala Cys Val Arg Thr Phe Tyr
Glu Thr Pro Leu Gln 100 105 110Leu Leu Glu Lys Val Lys Asn Val Phe
Asn Glu Thr Lys Asn Leu Leu 115 120 125Asp Lys Asp Trp Asn Ile Phe
Ser Lys Asn Cys Asn Asn Ser Phe Ala 130 135 140Glu Cys Ser Ser Gln
Gly His Glu Arg Gln Ser Glu Gly Ser145 150 15568222PRTHomo
sapiensmisc_feature(1)..(222)Human IL34 68Asn Glu Pro Leu Glu Met
Trp Pro Leu Thr Gln Asn Glu Glu Cys Thr1 5 10 15Val Thr Gly Phe Leu
Arg Asp Lys Leu Gln Tyr Arg Ser Arg Leu Gln 20 25 30Tyr Met Lys His
Tyr Phe Pro Ile Asn Tyr Lys Ile Ser Val Pro Tyr 35 40 45Glu Gly Val
Phe Arg Ile Ala Asn Val Thr Arg Leu Gln Arg Ala Gln 50 55 60Val Ser
Glu Arg Glu Leu Arg Tyr Leu Trp Val Leu Val Ser Leu Ser65 70 75
80Ala Thr Glu Ser Val Gln Asp Val Leu Leu Glu Gly His Pro Ser Trp
85 90 95Lys Tyr Leu Gln Glu Val Gln Thr Leu Leu Leu Asn Val Gln Gln
Gly 100 105 110Leu Thr Asp Val Glu Val Ser Pro Lys Val Glu Ser Val
Leu Ser Leu 115 120 125Leu Asn Ala Pro Gly Pro Asn Leu Lys Leu Val
Arg Pro Lys Ala Leu 130 135 140Leu Asp Asn Cys Phe Arg Val Met Glu
Leu Leu Tyr Cys Ser Cys Cys145 150 155 160Lys Gln Ser Ser Val Leu
Asn Trp Gln Asp Cys Glu Val Pro Ser Pro 165 170 175Gln Ser Cys Ser
Pro Glu Pro Ser Leu Gln Tyr Ala Ala Thr Gln Leu 180 185 190Tyr Pro
Pro Pro Pro Trp Ser Pro Ser Ser Pro Pro His Ser Thr Gly 195 200
205Ser Val Arg Pro Val Arg Ala Gln Gly Glu Gly Leu Leu Pro 210 215
2206925PRTHomo sapiensmisc_feature(1)..(25)Human acceptor A FR1
69Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ser1
5 10 15Ser Val Lys Val Ser Cys Lys Ala Ser 20 257014PRTHomo
sapiensmisc_feature(1)..(14)Human acceptor A FR2 70Trp Val Arg Gln
Ala Pro Gly Gln Gly Leu Glu Trp Met Gly1 5 107132PRTHomo
sapiensmisc_feature(1)..(32)Human acceptor A FR3 71Arg Val Thr Ile
Thr Ala Asp Lys Ser Thr Ser Thr Ala Tyr Met Glu1 5 10 15Leu Ser Ser
Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys Ala Arg 20 25
307211PRTHomo sapiensmisc_feature(1)..(11)Human acceptor A FR4
72Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser1 5 107325PRTHomo
sapiensmisc_feature(1)..(25)Human acceptor B FR1 73Gln Val Gln Leu
Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ser1 5 10 15Ser Val Lys
Val Ser Cys Lys Ala Ser 20 257414PRTHomo
sapiensmisc_feature(1)..(14)Human acceptor B FR2 74Trp Val Arg Gln
Ala Pro Gly Gln Gly Leu Glu Trp Met Gly1 5 107532PRTHomo
sapiensmisc_feature(1)..(32)Human acceptor B FR3 75Arg Val Thr Ile
Thr Ala Asp Lys Ser Thr Ser Thr Ala Tyr Met Glu1 5 10 15Leu Ser Ser
Leu Arg Ser Glu Asp
Thr Ala Val Tyr Tyr Cys Ala Arg 20 25 307611PRTHomo
sapiensmisc_feature(1)..(11)Human acceptor B FR4 76Trp Gly Gln Gly
Thr Leu Val Thr Val Ser Ser1 5 107725PRTHomo
sapiensmisc_feature(1)..(25)Human acceptor C FR1 77Gln Val Gln Leu
Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ser1 5 10 15Ser Val Lys
Val Ser Cys Lys Ala Ser 20 257814PRTHomo
sapiensmisc_feature(1)..(14)Human acceptor C FR2 78Trp Val Arg Gln
Ala Pro Gly Gln Gly Leu Glu Trp Met Gly1 5 107932PRTHomo
sapiensmisc_feature(1)..(32)Human acceptor C FR3 79Arg Val Thr Ile
Thr Ala Asp Lys Ser Thr Ser Thr Ala Tyr Met Glu1 5 10 15Leu Ser Ser
Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys Ala Arg 20 25
308011PRTHomo sapiensmisc_feature(1)..(11)Human acceptor C FR4
80Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser1 5 108123PRTHomo
sapiensmisc_feature(1)..(23)Human acceptor D FR1 81Glu Ile Val Leu
Thr Gln Ser Pro Ala Thr Leu Ser Leu Ser Pro Gly1 5 10 15Glu Arg Ala
Thr Leu Ser Cys 208215PRTHomo sapiensmisc_feature(1)..(15)Human
acceptor D FR2 82Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu
Leu Ile Tyr1 5 10 158332PRTHomo sapiensmisc_feature(1)..(32)Human
acceptor D FR3 83Gly Ile Pro Ala Arg Phe Ser Gly Ser Gly Ser Gly
Thr Asp Phe Thr1 5 10 15Leu Thr Ile Ser Ser Leu Glu Pro Glu Asp Phe
Ala Val Tyr Tyr Cys 20 25 308410PRTHomo
sapiensmisc_feature(1)..(10)Human acceptor D FR4 84Phe Gly Gly Gly
Thr Lys Val Glu Ile Lys1 5 108523PRTHomo
sapiensmisc_feature(1)..(23)Human acceptor E FR1 85Glu Ile Val Leu
Thr Gln Ser Pro Ala Thr Leu Ser Leu Ser Pro Gly1 5 10 15Glu Arg Ala
Thr Leu Ser Cys 208615PRTHomo sapiensmisc_feature(1)..(15)Human
acceptor E FR2 86Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu
Leu Ile Tyr1 5 10 158732PRTHomo sapiensmisc_feature(1)..(32)Human
acceptor E FR3 87Gly Ile Pro Ala Arg Phe Ser Gly Ser Gly Ser Gly
Thr Asp Phe Thr1 5 10 15Leu Thr Ile Ser Ser Leu Glu Pro Glu Asp Phe
Ala Val Tyr Tyr Cys 20 25 308810PRTHomo
sapiensmisc_feature(1)..(10)Human acceptor E FR4 88Phe Gly Gln Gly
Thr Lys Val Glu Ile Lys1 5 108923PRTHomo
sapiensmisc_feature(1)..(23)Human acceptor F FR1 89Glu Ile Val Leu
Thr Gln Ser Pro Ala Thr Leu Ser Leu Ser Pro Gly1 5 10 15Glu Arg Ala
Thr Leu Ser Cys 209015PRTHomo sapiensmisc_feature(1)..(15)Human
acceptor F FR2 90Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu
Leu Ile Tyr1 5 10 159132PRTHomo sapiensmisc_feature(1)..(32)Human
acceptor F FR3 91Gly Ile Pro Ala Arg Phe Ser Gly Ser Gly Ser Gly
Thr Asp Phe Thr1 5 10 15Leu Thr Ile Ser Ser Leu Glu Pro Glu Asp Phe
Ala Val Tyr Tyr Cys 20 25 309210PRTHomo
sapiensmisc_feature(1)..(10)Human acceptor F FR4 92Phe Gly Gln Gly
Thr Lys Val Glu Ile Lys1 5 1093719PRTMus
musculusmisc_feature(1)..(719)mCSF1R ECD-Fc 93Ala Pro Val Ile Glu
Pro Ser Gly Pro Glu Leu Val Val Glu Pro Gly1 5 10 15Glu Thr Val Thr
Leu Arg Cys Val Ser Asn Gly Ser Val Glu Trp Asp 20 25 30Gly Pro Ile
Ser Pro Tyr Trp Thr Leu Asp Pro Glu Ser Pro Gly Ser 35 40 45Thr Leu
Thr Thr Arg Asn Ala Thr Phe Lys Asn Thr Gly Thr Tyr Arg 50 55 60Cys
Thr Glu Leu Glu Asp Pro Met Ala Gly Ser Thr Thr Ile His Leu65 70 75
80Tyr Val Lys Asp Pro Ala His Ser Trp Asn Leu Leu Ala Gln Glu Val
85 90 95Thr Val Val Glu Gly Gln Glu Ala Val Leu Pro Cys Leu Ile Thr
Asp 100 105 110Pro Ala Leu Lys Asp Ser Val Ser Leu Met Arg Glu Gly
Gly Arg Gln 115 120 125Val Leu Arg Lys Thr Val Tyr Phe Phe Ser Pro
Trp Arg Gly Phe Ile 130 135 140Ile Arg Lys Ala Lys Val Leu Asp Ser
Asn Thr Tyr Val Cys Lys Thr145 150 155 160Met Val Asn Gly Arg Glu
Ser Thr Ser Thr Gly Ile Trp Leu Lys Val 165 170 175Asn Arg Val His
Pro Glu Pro Pro Gln Ile Lys Leu Glu Pro Ser Lys 180 185 190Leu Val
Arg Ile Arg Gly Glu Ala Ala Gln Ile Val Cys Ser Ala Thr 195 200
205Asn Ala Glu Val Gly Phe Asn Val Ile Leu Lys Arg Gly Asp Thr Lys
210 215 220Leu Glu Ile Pro Leu Asn Ser Asp Phe Gln Asp Asn Tyr Tyr
Lys Lys225 230 235 240Val Arg Ala Leu Ser Leu Asn Ala Val Asp Phe
Gln Asp Ala Gly Ile 245 250 255Tyr Ser Cys Val Ala Ser Asn Asp Val
Gly Thr Arg Thr Ala Thr Met 260 265 270Asn Phe Gln Val Val Glu Ser
Ala Tyr Leu Asn Leu Thr Ser Glu Gln 275 280 285Ser Leu Leu Gln Glu
Val Ser Val Gly Asp Ser Leu Ile Leu Thr Val 290 295 300His Ala Asp
Ala Tyr Pro Ser Ile Gln His Tyr Asn Trp Thr Tyr Leu305 310 315
320Gly Pro Phe Phe Glu Asp Gln Arg Lys Leu Glu Phe Ile Thr Gln Arg
325 330 335Ala Ile Tyr Arg Tyr Thr Phe Lys Leu Phe Leu Asn Arg Val
Lys Ala 340 345 350Ser Glu Ala Gly Gln Tyr Phe Leu Met Ala Gln Asn
Lys Ala Gly Trp 355 360 365Asn Asn Leu Thr Phe Glu Leu Thr Leu Arg
Tyr Pro Pro Glu Val Ser 370 375 380Val Thr Trp Met Pro Val Asn Gly
Ser Asp Val Leu Phe Cys Asp Val385 390 395 400Ser Gly Tyr Pro Gln
Pro Ser Val Thr Trp Met Glu Cys Arg Gly His 405 410 415Thr Asp Arg
Cys Asp Glu Ala Gln Ala Leu Gln Val Trp Asn Asp Thr 420 425 430His
Pro Glu Val Leu Ser Gln Lys Pro Phe Asp Lys Val Ile Ile Gln 435 440
445Ser Gln Leu Pro Ile Gly Thr Leu Lys His Asn Met Thr Tyr Phe Cys
450 455 460Lys Thr His Asn Ser Val Gly Asn Ser Ser Gln Tyr Phe Arg
Ala Val465 470 475 480Ser Leu Gly Gln Ser Lys Gln Glu Pro Lys Ser
Ser Asp Lys Thr His 485 490 495Thr Cys Pro Pro Cys Pro Ala Pro Glu
Leu Leu Gly Gly Pro Ser Val 500 505 510Phe Leu Phe Pro Pro Lys Pro
Lys Asp Thr Leu Met Ile Ser Arg Thr 515 520 525Pro Glu Val Thr Cys
Val Val Val Asp Val Ser His Glu Asp Pro Glu 530 535 540Val Lys Phe
Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys545 550 555
560Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser
565 570 575Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu
Tyr Lys 580 585 590Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile
Glu Lys Thr Ile 595 600 605Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro
Gln Val Tyr Thr Leu Pro 610 615 620Pro Ser Arg Asp Glu Leu Thr Lys
Asn Gln Val Ser Leu Thr Cys Leu625 630 635 640Val Lys Gly Phe Tyr
Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn 645 650 655Gly Gln Pro
Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser 660 665 670Asp
Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg 675 680
685Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu
690 695 700His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly
Lys705 710 71594327PRTArtificial sequenceSynthetic Human IgG4 S241P
94Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Cys Ser Arg1
5 10 15Ser Thr Ser Glu Ser Thr Ala Ala Leu Gly Cys Leu Val Lys Asp
Tyr 20 25 30Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu
Thr Ser 35 40 45Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly
Leu Tyr Ser 50 55 60Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu
Gly Thr Lys Thr65 70 75 80Tyr Thr Cys Asn Val Asp His Lys Pro Ser
Asn Thr Lys Val Asp Lys 85 90 95Arg Val Glu Ser Lys Tyr Gly Pro Pro
Cys Pro Pro Cys Pro Ala Pro 100 105 110Glu Phe Leu Gly Gly Pro Ser
Val Phe Leu Phe Pro Pro Lys Pro Lys 115 120 125Asp Thr Leu Met Ile
Ser Arg Thr Pro Glu Val Thr Cys Val Val Val 130 135 140Asp Val Ser
Gln Glu Asp Pro Glu Val Gln Phe Asn Trp Tyr Val Asp145 150 155
160Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Phe
165 170 175Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His
Gln Asp 180 185 190Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser
Asn Lys Gly Leu 195 200 205Pro Ser Ser Ile Glu Lys Thr Ile Ser Lys
Ala Lys Gly Gln Pro Arg 210 215 220Glu Pro Gln Val Tyr Thr Leu Pro
Pro Ser Gln Glu Glu Met Thr Lys225 230 235 240Asn Gln Val Ser Leu
Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp 245 250 255Ile Ala Val
Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys 260 265 270Thr
Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser 275 280
285Arg Leu Thr Val Asp Lys Ser Arg Trp Gln Glu Gly Asn Val Phe Ser
290 295 300Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln
Lys Ser305 310 315 320Leu Ser Leu Ser Leu Gly Lys 32595107PRTHomo
sapiensmisc_feature(1)..(107)Human Igk 95Arg Thr Val Ala Ala Pro
Ser Val Phe Ile Phe Pro Pro Ser Asp Glu1 5 10 15Gln Leu Lys Ser Gly
Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe 20 25 30Tyr Pro Arg Glu
Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln 35 40 45Ser Gly Asn
Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser 50 55 60Thr Tyr
Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu65 70 75
80Lys His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser
85 90 95Pro Val Thr Lys Ser Phe Asn Arg Gly Glu Cys 100 105
* * * * *