Antikine antibodies that bind to multiple cc chemokines

Abstract

An antikine antibody binds to two, three, four, five or more CC chemokines, such as RANTES/CCL5, MIP-1α/CCL3, MIP-1β/CCL4, or MCP-1/CCL2. Methods for affinity maturation and humanization of antikine antibodies as well as the production of hybridoma cell lines producing antikine antibodies by sequential immunization are also disclosed.

1 . A method of treating a disease, disorder or condition mediated by one or more CC chemokines comprising administering to a subject in need thereof an isolated antikine antibody or an antigen binding fragment thereof that binds to CCL3/MIP-1α, CCL4/MIP-1β, and CCL4/RANTES, wherein the antibody comprises the three heavy chain CDRs (HCDR1, HCDR2, and HCDR3) contained within the heavy chain variable region sequence selected from SEQ ID NOS:2, 12, 22, 32, 42, 52, or 62. 
     
     
         2 . The method of  claim 1 , wherein the isolated antibody or antigen binding fragment further comprises the three light chain CDRs (LCDR1, LCDR2, and LCDR3) contained within the light chain variable region of selected from SEQ ID NOS:7, 17, 27, 37, 47, 57, or 67. 
     
     
         3 . The method of  claim 1 , wherein the isolated antibody or antigen binding fragment thereof is a chimeric monoclonal antibody or an antigen binding fragment thereof. 
     
     
         4 . The method of  claim 1 , wherein the isolated antibody or antigen binding fragment thereof is a humanized monoclonal antibody or an antigen binding fragment thereof. 
     
     
         5 . The method of  claim 1 , wherein the isolated antibody or antigen binding fragment thereof is a full-length monoclonal antibody. 
     
     
         6 . The method of  claim 1 , wherein the isolated antigen binding fragment thereof is a Fab′ or F(ab′)2. 
     
     
         7 . The method of  claim 1 , wherein the isolated antibody or antigen binding fragment thereof comprises a heavy chain variable region sequence comprising SEQ ID NO:2 and a light chain variable region sequence comprising SEQ ID NO:7. 
     
     
         8 . The method of  claim 1 , wherein the isolated antibody or antigen binding fragment thereof comprises a heavy chain variable region sequence comprising SEQ ID NO:12 and a light chain variable region sequence comprising SEQ ID NO:17. 
     
     
         9 . The method of  claim 1 , wherein the isolated antibody or antigen binding fragment thereof comprises a heavy chain variable region sequence comprising SEQ ID NO:22 and a light chain variable region sequence comprising SEQ ID NO:27. 
     
     
         10 . The method of  claim 1 , wherein the antibody or antigen binding fragment thereof comprises a heavy chain variable region sequence comprising SEQ ID NO:32 and a light chain variable region sequence comprising SEQ ID NO:37. 
     
     
         11 . The method of  claim 1 , wherein the antibody or antigen binding fragment thereof comprises a heavy chain variable region sequence comprising SEQ ID NO:42 and a light chain variable region sequence comprising SEQ ID NO:47. 
     
     
         12 . The method of  claim 1 , wherein the antibody or antigen binding fragment thereof comprises a heavy chain variable region sequence comprising SEQ ID NO:52 and a light chain variable region sequence comprising SEQ ID NO:57. 
     
     
         13 . The method of  claim 1 , wherein the isolated antibody or antigen binding fragment thereof is produced by a hybridoma selected from the group consisting of hybridoma cell line 3C12F deposited at the ATCC as PTA-11261, 7D1G deposited at the ATCC as PTA-11257, 7D12A deposited at the ATCC as PTA-11259, 18V4F deposited at the ATCC as PTA-11260 or 18P7E deposited at the ATCC as PTA-11258 or which competitively blocks binding of an antibody produced by hybridoma cell line 3C12F deposited at the ATCC as PTA-11261, 7D1G deposited at the ATCC as PTA-11257, 7D12A deposited at the ATCC as PTA-11259, 18V4F deposited at the ATCC as PTA-11260 and 18P7E deposited at the ATCC as PTA-11258. 
     
     
         14 . The method of  claim 1 , wherein the isolated antibody or antigen binding fragment thereof has a binding affinity of at least 20 nM for at least three different CC chemokines, wherein at least one CC chemokine is CCL3/MIP-1α, CCL4/MIP-1β, or CCL5/RANTES. 
     
     
         15 . The method of  claim 1 , wherein the isolated antibody or antigen binding fragment thereof binds to at least four different CC chemokines. 
     
     
         16 . The method of  claim 1 , wherein the isolated antibody or antigen binding fragment thereof does not bind to MCP-1, MCP-2 or MCP-3. 
     
     
         17 . The method of  claim 1 , wherein the isolated antibody or antigen binding fragment thereof binds to at least three different CC chemokines selected from the group consisting of CCL2/MCP-1, CCL14/HCC-1, CCL15/HCC-2, CCL18/PARC, and CCL23/MPIF-1. 
     
     
         18 . The method of  claim 1 , wherein the isolated antibody or antigen binding fragment thereof binds to at least one determinant of CCL2/MCP-1, CCL3/MIP-1α, CCL4/MIP-1β, CCL5/RANTES, CCL14/HCC-1, CCL15/HCC-2, CCL18/PARC, and CCL23/MPIF-1 which determinant is located between the CC residues of said CC-chemokine and the last C residue of said chemokine. 
     
     
         19 . The method of  claim 1 , wherein the isolated antibody or antigen binding fragment thereof binds to at least one determinant of CCL2/MCP-1, CCL3/MIP-1α, CCL4/MIP-1β, CCL5/RANTES, CCL14/HCC-1, CCL15/HCC-2, CCL18/PARC, and CCL23/MPIF-1 which is located in the N-loop, 30's loop, or 40's loop of said CC chemokine. 
     
     
         20 . The method of  claim 1 , wherein the isolated antibody or antigen binding fragment thereof binds to at least one determinant within CC receptor binding residues of a CC chemokine. 
     
     
         21 . The method of  claim 1 , wherein the isolated antibody or antigen binding fragment thereof neutralizes the chemotactic activity of a CC chemokine to which it binds. 
     
     
         22 . The method of  claim 1 , wherein the isolated antibody or antigen binding fragment thereof binds to at least five CC chemokines selected from the group consisting of CCL3/MIP-1α, CCL4/MIP-1β, CCL5/RANTES, CCL15/HCC-2, and CCL23/MPIF-1, and which does not substantially bind to CCL2/MCP-1. 
     
     
         23 . The method of  claim 22 , wherein the isolated antibody or antigen binding fragment thereof binds to a determinant in the N loop, 30's loop, or 40's loop of at least one of said CC chemokines. 
     
     
         24 . The method of  claim 22 , wherein the isolated antibody or antigen binding fragment thereof binds to at least one antigenic determinant of CCL3/MIP-1α located within residues 11-15 (CCFSY), residues 17-24 (SRQIPQNF), residues 34-35 (QC), or residues 57-67 (EWVQKYVSDLE) of SEQ ID NO: 71;
 binds to at least one antigenic determinant of CCL4/MIP-1β located within residues 11-15 (CCFSY), residues 17-24 (ARKLPHNF), residues 34-35 (LC), or residues 57-67 (SWVQEYVYDLE) of SEQ ID NO: 72; 
 binds to at least one antigenic determinant of CCL5/RANTES located within residues 10-14 (CCFAY), residues 16-23 (ARPLPRAH), residues 33-34 (KC), or residues 56-66 (KWVREYINSLE) of SEQ ID NO: 73. 
 
     
     
         25 . The method of  claim 1 , wherein a disease, disorder or condition mediated by one or more CC chemokines is an inflammatory disease or a disease mediated by or associated with inflammation. 
     
     
         26 . The method of  claim 1 , wherein a disease, disorder or condition mediated by one or more CC chemokines is an autoimmune disease. 
     
     
         27 . The method of  claim 1 , wherein the antibody or antigen binding fragment thereof has been conjugated to an effector moiety, targeting moiety, heterologous protein, toxin, enzyme, cytokine, chemical tag, radiological tag, a substance that increases its biological half-life or a substance that increases its biological availability. 
     
     
         28 . The method of  claim 1 , wherein the antibody or antigen binding fragment thereof further comprises a carrier or excipient. 
     
     
         29 . A kit for use in a therapeutic or diagnostic procedure that comprises one more isolated antikine antibody or an antigen binding fragment thereof that binds to CCL3/MIP-1α, CCL4/MIP-1β, and CCL4/RANTES, wherein the antibody comprises the three heavy chain CDRs (HCDR1, HCDR2, and HCDR3) contained within the heavy chain variable region sequence selected from SEQ ID NOS:2, 12, 22, 32, 42, 52, or 62, at least one positive control antibody, at least one negative control antibody, at least one CC chemokine bound by said one or more antikine antibodies, and at least one control chemokine not bound by the antikine antibody.