Monoclonal Antibodies Against PCBP-1 Antigens, and Uses Therefor

Abstract

The present invention provides and includes monoclonal antibodies (MoAbs or mAbs) specific or preferentially selective for PCBP-1 antigens, hybridoma lines that secrete these PCBP-1 antibodies or antibody fragments, and the use of such antibodies and antibody fragments to detect PCBP-1 antigens, particularly those expressed by cancer cells. The present invention also includes antibodies that are specific for or show preferential binding to a soluble form of PCBP-1. The present invention further includes chimeric and humanized antibodies, processes for producing monoclonal, chimeric, and humanized antibodies using recombinant DNA technology, and their therapeutic uses, particularly in the treatment of cancer. The present invention further includes methods and kits for the immunodetection and immunotherapy of cells for samples which express PCBP-1 antigens.

1 . An antibody specific for a PCBP-1 antigen, comprising the heavy chain CDR antigen binding site sequences CDR1, CDR2, and CDR3 selected from the group consisting of SEQ ID NO: 45, SEQ ID NO: 46 and SEQ ID NO: 47, and the light chain CDR antigen binding site sequences CDR1, CDR2, and CDR3 selected from the group consisting of SEQ ID NO: 48, SEQ ID NO: 49 and SEQ ID NO: 50. 
     
     
         2 . An antibody specific for a PCBP-1 antigen, comprising one or more of the heavy chain CDR antigen binding site sequences selected from the group consisting of SEQ ID NO: 45, SEQ ID NO: 46 and SEQ ID NO: 47, and one or more of the light chain CDR antigen binding site sequences selected from the group consisting of SEQ ID NO: 48, SEQ ID NO: 49 and SEQ ID NO: 50. 
     
     
         3 . An antibody according to  claim 1  wherein said PCBP-1 antigen is a soluble protein having a molecular weight of about 40 kilodaltons as measured by gradient polyacrylamide gel electrophoresis. 
     
     
         4 . An antibody according to  claim 1  immobilized on a solid phase. 
     
     
         5 . An antibody according to  claim 1  which is detectably labeled. 
     
     
         6 . An antibody according to  claim 1  conjugated to a cytotoxic radionuclide. 
     
     
         7 . An antibody according to  claim 1  conjugated to a cytotoxic drug. 
     
     
         8 . An antibody according to  claim 1  conjugated to a cytotoxic protein. 
     
     
         9 . An antibody according to  claim 2  immobilized on a solid phase. 
     
     
         10 . An antibody according to  claim 2  which is detectably labeled. 
     
     
         11 . An antibody according to  claim 2  conjugated to a cytotoxic radionuclide. 
     
     
         12 . An antibody according to  claim 2  conjugated to a cytotoxic drug. 
     
     
         13 . An antibody according to  claim 2  conjugated to a cytotoxic protein. 
     
     
         14 . A composition comprising an antibody according to  claim 6  in combination with a pharmaceutically acceptable carrier. 
     
     
         15 . An antibody capable of preferentially binding to a soluble form of PCBP-1 antigen. 
     
     
         16 . An antibody according to  claim 15 , wherein said preferential binding is relative to a membrane form of PCBP-1. 
     
     
         17 . An isolated DNA sequence which encodes the heavy chain of an antibody molecule, wherein said antibody molecule has specificity for PCBP-1 and wherein the variable domain of said heavy chain comprises a CDR having the antigen binding site sequences CDR1, CDR2, and CDR3, selected from the group consisting of SEQ ID NO: 45, SEQ ID NO: 46 and SEQ ID NO: 47. 
     
     
         18 . An isolated DNA sequence which encodes the light chain of an antibody molecule, wherein said antibody molecule has specificity for PCBP-1 and further wherein the variable domain of said light chain comprises a CDR having the antigen binding site sequences CDR1, CDR2, and CDR3, selected from the group consisting of SEQ ID NO: 48, SEQ ID NO: 49 and SEQ ID NO: 50. 
     
     
         19 . A vector containing the DNA sequence of  claim 17 . 
     
     
         20 . A vector containing the DNA sequence of  claim 18 . 
     
     
         21 . A host cell transformed with the vector of  claim 19 . 
     
     
         22 . A host cell transformed with the vector of  claim 20 . 
     
     
         23 . A process for the production of an antibody molecule having specificity for PCBP-1, comprising culturing the host cell of  claim 21  and isolating the antibody molecule. 
     
     
         24 . A process for the production of an antibody molecule having specificity for PCBP-1, comprising culturing the host cell of  claim 22  and isolating the antibody molecule. 
     
     
         25 . An immunoassay for detecting a PCBP-1 antigen which binds to a monoclonal antibody having the one or more of the heavy chain CDR antigen binding site sequences selected from the group consisting of SEQ ID NO: 45, SEQ ID NO: 46 and SEQ ID NO: 47, and one or more of the light chain CDR antigen binding site sequences selected from the group consisting of SEQ ID NO: 48, SEQ ID NO: 49 and SEQ ID NO: 50, comprising: (a) contacting said sample with an effective binding amount of the antibody according to  claim 1 ; and (b) detecting said antigen by detecting the binding of the antibody to the PCBP-1 antigen. 
     
     
         26 . An immunoassay for detecting a PCBP-1 antigen which binds to a monoclonal antibody having the one or more of the heavy chain CDR antigen binding site sequences selected from the group consisting of SEQ ID NO: 45, SEQ ID NO: 46 and SEQ ID NO: 47, and one or more of the light chain CDR antigen binding site sequences selected from the group consisting of SEQ ID NO: 48, SEQ ID NO: 49 and SEQ ID NO: 50, comprising: (a) contacting said sample with an effective binding amount of the antibody according to  claim 2 ; and (b) detecting said antigen by detecting the binding of the antibody to the PCBP-1 antigen. 
     
     
         27 . The immunoassay of  claim 25 , wherein the assay is used to detect cancer cells expressing a PCBP-1 antigen, said cancer cells being selected from the group consisting of breast, ovarian, cervical, prostate, colon, stomach, kidney, liver, head, neck, lung, blood, pancreatic, skin, testicular, thyroid and brain cancers. 
     
     
         28 . The immunoassay of  claim 26 , wherein the assay is used to detect cancer cells expressing a PCBP-1 antigen, said cancer cells being selected from the group consisting of breast, ovarian, cervical, prostate, colon, stomach, kidney, liver, head, neck, lung, blood, pancreatic, skin, testicular, thyroid and brain cancers. 
     
     
         29 . A kit for the immunohistochemical detection of carcinoma comprising: (a) a monoclonal antibody having one or more of the heavy chain CDR antigen binding site sequences selected from the group consisting of SEQ ID NO: 45, SEQ ID NO: 46 and SEQ ID NO: 47, and one or more of the light chain CDR antigen binding site sequences selected from the group consisting of SEQ ID NO: 48, SEQ ID NO: 49 and SEQ ID NO: 50; and (b) a secondary antibody conjugated to a detectable label. 
     
     
         30 . An immunoassay for detecting a PCBP-1 antigen which binds to a monoclonal antibody having one or more of the heavy chain CDR antigen binding site sequences selected from the group consisting of SEQ ID NO: 45, SEQ ID NO: 46 and SEQ ID NO: 47, and one or more of the light chain CDR antigen binding site sequences selected from the group consisting of SEQ ID NO: 48, SEQ ID NO: 49 and SEQ ID NO: 50 in a sample comprising: (a) contacting said sample with the antibody; and (b) detecting said antigen by detecting the binding of said antibody to the protein antigen. 
     
     
         31 . A method for diagnosing cancer in humans comprising: (a) removing a specimen from a patient suspected of having a cancer; (b) contacting the specimen with a monoclonal antibody having one or more of the heavy chain CDR antigen binding site sequences selected from the group consisting of SEQ ID NO: 45, SEQ ID NO: 46 and SEQ ID NO: 47, and one or more of the light chain CDR antigen binding site sequences selected from the group consisting of SEQ ID NO: 48, SEQ ID NO: 49 and SEQ ID NO: 50, thereby forming antigen-antibody complexes in said specimen; (c) labeling the specimen with a label specific for the antigen-antibody complex; and (d) detecting the presence of the antigen-antibody complex by detecting the label. 
     
     
         32 . The method of  claim 31 , wherein the method is performed in vivo. 
     
     
         33 . The method of  claim 31 , wherein the method is performed in vitro. 
     
     
         34 . The method of  claim 31 , wherein the cancer being diagnosed is selected from the group consisting of breast, ovarian, cervical, prostate, colon, stomach, kidney, liver, head, neck, lung, blood, pancreatic, skin, testicular, thyroid and brain cancer. 
     
     
         35 . A method of determining the status of a cell in a sample comprising:
 (a) obtaining said sample from a patient;   (b) contacting said sample with an antibody capable of preferentially detecting a soluble form of PCBP-1 antigen; and   (c) determining the quantity of said antigen.   
     
     
         36 . A method of determining the status of a cell in a sample comprising:
 (a) obtaining said sample from a patient;   (b) contacting said sample with an antibody capable of preferentially detecting a soluble form of PCBP-1 antigen; and   (c) determining the localization of said antigen.   
     
     
         37 . The method of  claim 36 , further comprising contacting said sample with an antibody capable of preferentially detecting a non-soluble form of PCBP-1 antigen. 
     
     
         38 . A method of determining the status of cell in a sample by:
 (a) obtaining said sample;   (b) contacting said sample with an antibody capable of preferentially detecting a soluble form of PCBP-1 antigen;   (c) determining quantity or localization of said antigen.   
     
     
         39 . A method according to  claim 38 , further comprising contacting said sample with an antibody capable of preferentially detecting a non-soluble form of PCBP-1 antigen. 
     
     
         40 . An antibody capable of binding to a soluble form of PCBP-1 with a specific affinity of between 10 −8  M and 10 −11  M. 
     
     
         41 . An antibody capable of binding a soluble form of PCBP-1 in a cell. 
     
     
         42 . An antibody capable of selectively reducing the activity of a soluble PCBP-1 in a cell. 
     
     
         43 . An antibody according to  claim 15 , wherein said preferential binding is relative to a membrane form of PCBP-1. 
     
     
         44 . An antibody according to  claim 43 , wherein said membrane is a nuclear membrane. 
     
     
         45 . An antibody according to  claim 41 , wherein said cell is in vitro. 
     
     
         46 . An antibody according to  claim 42 , wherein said cell is in vitro. 
     
     
         47 . An antibody capable of recognizing an epitope selected from the group consisting of SEQ ID NOs: 1-14, and fragments thereof. 
     
     
         48 . An antibody capable of preferentially binding to a secreted form of PCBP-1 antigen. 
     
     
         49 . An antibody according to  claim 16 , wherein said membrane is a nuclear membrane.