HUMANIZED ANTIBODIES AGAINST MONOCYTE CHEMOTACTIC PROTEINS
The invention provides humanized antibodies that bind to a plurality of b-chemokines, particularly monocyte chemotactic proteins MCP-1, MCP-2 and MCP-3. The invention also provides therapeutic reagents and methods of treating disorders associated with detrimental MCP activity.
1 - 69 . (canceled)
70 . A method of treating a subject suffering from a disorder selected from glomerulonephritis, scleroderma, cirrhosis, multiple sclerosis, lupus nephritis, atherosclerosis, inflammatory bowel disease or rheumatoid arthritis, comprising administering to the subject a humanized immunoglobulin or antigen-binding fragment thereof comprising a) heavy chain complementarity determining regions as set forth in SEQ ID NO:29, SEQ ID NO:30, and SEQ ID NO:31, and b) light chain complementarity determining regions as set forth in SEQ ID NO:32, SEQ ID NO:33, and SEQ ID NO:34, wherein the remainder of the heavy and light chains are from a human immunoglobulin, wherein the immunoglobulin or antigen-binding fragment thereof binds to MCP-1, MCP-2, or MCP-3, and wherein the humanized immunoglobulin or antigen-binding fragment thereof is administered in an amount sufficient to reduce the activity of at least one of MCP-1, MCP-2, or MCP-3.
71 . The method of claim 70 , wherein the heavy chain further comprises at least one variable region framework residue selected from L27, I29, and T73 (Kabat numbering convention) from the monoclonal antibody 11K2 heavy chain set forth as SEQ ID NO:27.
72 . The method of claim 70 , wherein the heavy chain further comprises variable region framework residues L27, I29, and T73 (Kabat numbering convention) from the monoclonal antibody 11K2 heavy chain set forth as SEQ ID NO:27.
73 . The method of claim 70 , wherein the heavy chain further comprises at least one variable region framework residue selected from N28, K30, I48, and A67 (Kabat numbering convention) from the monoclonal antibody 11K2 heavy chain as set forth in SEQ ID NO:27.
74 . The method of claim 70 , wherein the heavy chain further comprises variable region framework residues N28, K30, I48, and A67 (Kabat numbering convention) from the monoclonal antibody 11K2 heavy chain as set forth in SEQ ID NO:27.
75 . The method of claim 70 , wherein the light chain further comprises at least one variable region framework residue selected from S49 and Y71 (Kabat numbering convention) from the monoclonal antibody 11K2 light chain set forth as SEQ ID NO: 28.
76 . The method of claim 70 , wherein the light chain further comprises variable region framework residues S49 and Y71 (Kabat numbering convention) from the monoclonal antibody 11K2 light chain as set forth in SEQ ID NO:28.
77 . The method of claim 76 , wherein the light chain further comprises variable region framework residue K69 (Kabat numbering convention) from the monoclonal antibody 11K2 light chain as set forth in SEQ ID NO:28.
78 . The method of claim 70 , wherein the heavy chain further comprises at least one variable region framework residue from the monoclonal antibody 11K2 heavy chain set forth as SEQ ID NO: 27, wherein the residue is selected from the group consisting of L27, N28, I29, K30, I48, A67, and T73 (Kabat numbering).
79 . The method of claim 78 , wherein the heavy chain comprises variable region framework residues L27, N28, I29, K30, I48, A67, and T73 (Kabat numbering convention) from the monoclonal antibody 11K2 heavy chain as set forth in SEQ ID NO:27.
80 . The method of claim 76 , wherein the heavy chain comprises at least one variable region framework residues L27, I29, and T73 (Kabat numbering convention) from the monoclonal antibody 11K2 heavy chain as set forth in SEQ ID NO:27.
81 . The method of claim 79 , wherein the light chain comprises at least one variable region framework residue selected from S49, K69, and Y71 (Kabat numbering convention) from the monoclonal antibody 11K2 light chain set forth as SEQ ID NO: 28.
82 . The method of claim 70 , wherein the heavy chain comprises at least one variable region framework residue selected from L27, N28, I29, K30, I48, A67 and T73 (Kabat numbering convention) from the monoclonal antibody 11K2 heavy chain set forth as SEQ ID NO: 27, and wherein the light chain comprises at least one variable region framework residue selected from S49, K69, and Y71 (Kabat numbering convention) from the monoclonal antibody 11K2 light chain set forth as SEQ ID NO: 28, wherein the remainder of the heavy and light chains are from a human immunoglobulin.
83 . The method of claim 82 , wherein the heavy chain comprises variable region framework residues L27, I29, and T73, and the light chain comprises variable region framework residues S49 and Y71.
84 . The method of claim 83 , wherein the light chain further comprises variable region framework residue K69.
85 . The method of claim 82 , wherein the heavy chain comprises variable region framework residues L27, N28, I29, K30, I48, A67 and T73, and the light chain comprises variable region framework residues S49 and Y71.
86 . The method of claim 70 , wherein the humanized immunoglobulin or antigen-binding fragment thereof is modified by reducing or eliminating at least one potential glycosylation site.
87 . The method of claim 70 , wherein the immunoglobulin or antigen-binding fragment thereof specifically binds to MCP-1 with a binding affinity of at least 10 −9 M.
88 . The method of claim 70 , wherein the immunoglobulin or antigen-binding fragment thereof specifically binds to MCP-1 with a binding affinity of at least 10 −10 M.
89 . The method of claim 70 , wherein the immunoglobulin or antigen-binding fragment thereof specifically binds to MCP-1 with a binding affinity of at least 10 −10 M.
90 . The method of claim 70 , wherein the immunoglobulin or antigen-binding fragment thereof specifically binds to MCP-1 with a binding affinity of 1 pM or less or 0.4 pM to about 0.7 pM.
91 . The method of claim 87 , wherein the immunoglobulin or antigen-binding fragment thereof further binds to MCP-2 with a binding affinity of at least 10 −7 M.
92 . The method of claim 87 , wherein the immunoglobulin or antigen-binding fragment of thereof further binds to MCP-2 with a binding affinity of at least 10 −8 M.
93 . The method of claim 87 , wherein the immunoglobulin or antigen-binding fragment thereof further binds to MCP-2 with a binding affinity of at least 10 −9 M.
94 . The method of claim 70 , wherein the immunoglobulin or antigen-binding fragment thereof binds to MCP-2.
95 . The method of claim 70 , wherein the immunoglobulin or antigen-binding fragment thereof specifically binds to MCP-2 with a binding affinity of at least 10 −7 M.
96 . The method of claim 70 , wherein the immunoglobulin or antigen-binding fragment thereof specifically binds to MCP-2 with a binding affinity of at least 10 −8 M.
97 . The method of claim 70 , wherein the immunoglobulin or antigen-binding fragment thereof specifically binds to MCP-2 with a binding affinity of at least 10 −9 M.
98 . The method of claim 70 , wherein the immunoglobulin or antigen-binding fragment thereof specifically binds to MCP-1 and MCP-2.
99 . The method of claim 70 , wherein the immunoglobulin or antigen-binding fragment thereof binds to an epitope within MCP-1, MCP-2, and MCP-3.
100 . The method of claim 70 , wherein the immunoglobulin comprises the same heavy and light chain polypeptide sequences as an antibody produced by clone 3F2 (ATCC patent deposit designation PTA-5308).
101 . The method of claim 70 , wherein the immunoglobulin heavy chain isotype is gamma 1.
102 . The method of claim 70 , wherein the immunoglobulin is modified by reducing or eliminating at least one potential glycosylation site.
103 . The method of claim 70 , wherein the immunoglobulin or antigen-binding fragment thereof is modified by conjugation to a carrier selected from polyethylene glycol and albumen.
104 . The method of claim 70 , wherein the constant region of the immunoglobulin or antigen-binding fragment thereof is modified to reduce at least one constant region-mediated biological effector function relative to an unmodified antibody.
105 . The method of claim 70 , wherein the antigen-binding fragment is selected from an Fab, an Fab′, an F(ab) 2 , and an Fv.
106 . The method of claim 105 , wherein the antigen-binding fragment is an Fab.
107 . The method of claim 70 , wherein the immunoglobulin or antigen-binding fragment thereof inhibits MCP-induced chemotaxis in the subject.
108 . The method of claim 107 , wherein the immunoglobulin or antigen-binding fragment thereof inhibits MCP-1-induced chemotaxis, MCP-2-induced chemotaxis, or both MCP-1-induced and MCP-2-induced chemotaxis in the subject.