Cytotoxicity mediation of cells evidencing surface expression of TROP-2

Abstract

The present invention relates to a method for producing cancerous disease modifying antibodies using a novel paradigm of screening. By segregating the anti-cancer antibodies using cancer cell cytotoxicity as an end point, the process makes possible the production of anti-cancer antibodies for therapeutic and diagnostic purposes. The antibodies can be used in aid of staging and diagnosis of a cancer, and can be used to treat primary tumors and tumor metastases. The anti-cancer antibodies can be conjugated to toxins, enzymes, radioactive compounds, cytokines, interferons, target or reporter moieties and hematogenous cells.

1 . A humanized antibody of the isolated monoclonal antibody produced by the hybridoma deposited with the IDAC as accession number 141205-05 or an antigen binding fragment produced from said humanized antibody. 
     
     
         2 . A chimeric antibody of the isolated monoclonal antibody produced by the hybridoma deposited with the IDAC as accession number 141205-05 or an antigen binding fragment produced from said chimeric antibody. 
     
     
         3 . A method for initiating antibody induced cytotoxicity of cancerous cells in a tissue sample selected from a human pancreatic, ovarian, prostate or colon tumor comprising:
 providing a tissue sample from said human pancreatic, ovarian, prostate or colon tumor   providing the isolated monoclonal antibody produced by the hybridoma deposited with the IDAC as accession number 141205-05, the humanized antibody of the isolated monoclonal antibody produced by the hybridoma deposited with the IDAC as accession number 141205-05, the chimeric antibody of the isolated monoclonal antibody produced by the hybridoma deposited with the IDAC as accession number 141205-05, or an antigen binding fragment of any of the foregoing, characterized by an ability to competitively inhibit binding of said isolated monoclonal antibody to its target antigen; and   contacting said isolated monoclonal antibody, said humanized antibody, said chimeric antibody or said antigen binding fragment thereof with said tissue sample;   wherein binding of said isolated monoclonal antibody, said humanized antibody, said chimeric antibody or said antigen binding fragment thereof with said tissue sample induces cytotoxicity.   
     
     
         4 . An antigen binding fragment of the humanized antibody of  claim 1 . 
     
     
         5 . An antigen binding fragment of the chimeric antibody of  claim 2 . 
     
     
         6 . A method of reduction of a human pancreatic, ovarian, prostate or colon tumor susceptible to antibody induced cytotoxicity in a mammal, wherein said human pancreatic, ovarian, prostate or colon tumor expresses at least one epitope of an antigen which specifically binds to the isolated monoclonal antibody produced by the hybridoma deposited with the IDAC as accession number 141205-05 or an antigen binding fragment thereof, which antigen binding fragment is characterized by an ability to competitively inhibit binding of said isolated monoclonal antibody to its target antigen, comprising administering to said mammal said monoclonal antibody or said antigen binding fragment thereof in an amount effective to result in a reduction of said mammal's pancreatic, ovarian, prostate or colon tumor burden. 
     
     
         7 . The method of  claim 6  wherein said isolated monoclonal antibody is conjugated to a cytotoxic moiety. 
     
     
         8 . The method of  claim 7  wherein said cytotoxic moiety is a radioactive isotope. 
     
     
         9 . The method of  claim 6  wherein said isolated monoclonal antibody or antigen binding fragment thereof activates complement. 
     
     
         10 . The method of  claim 6  wherein said isolated monoclonal antibody or antigen binding fragment thereof mediates antibody dependent cellular cytotoxicity. 
     
     
         11 . The method of  claim 6  wherein said isolated monoclonal antibody is a humanized antibody of the isolated monoclonal antibody produced by the hybridoma deposited with the IDAC as accession number 141205-05. 
     
     
         12 . The method of  claim 6  wherein said isolated monoclonal antibody is a chimeric antibody of the isolated monoclonal antibody produced by the hybridoma deposited with the IDAC as accession number 141205-05. 
     
     
         13 . A monoclonal antibody which specifically binds to the same epitope or epitopes as the isolated monoclonal antibody produced by the hybridoma deposited with the IDAC as accession number 141205-05. 
     
     
         14 . A method of reduction of a human pancreatic, ovarian, prostate or colon tumor in a mammal, wherein said human tumor expresses at least one epitope of an antigen which specifically binds to the isolated monoclonal antibody produced by the hybridoma deposited with the IDAC as accession number 141205-05 or an antigen binding fragment thereof, which antigen binding fragment is characterized by an ability to competitively inhibit binding of said isolated monoclonal antibody to its target antigen, comprising administering to said mammal said monoclonal antibody or antigen binding fragment thereof in an amount effective to result in a reduction of said mammal's pancreatic, ovarian, prostate or colon tumor burden. 
     
     
         15 . The method of  claim 14  wherein said isolated monoclonal antibody is conjugated to a cytotoxic moiety. 
     
     
         16 . The method of  claim 15  wherein said cytotoxic moiety is a radioactive isotope. 
     
     
         17 . The method of  claim 14  wherein said isolated monoclonal antibody or antigen binding fragment thereof activates complement. 
     
     
         18 . The method of  claim 14  wherein said isolated monoclonal antibody or antigen binding fragment thereof mediates antibody dependent cellular cytotoxicity. 
     
     
         19 . The method of  claim 14  wherein said isolated monoclonal antibody is a humanized antibody of the isolated monoclonal antibody produced by the hybridoma deposited with the IDAC as accession number 141205-05. 
     
     
         20 . The method of  claim 14  wherein said isolated monoclonal antibody is a chimeric antibody of the isolated monoclonal antibody produced by the hybridoma deposited with the IDAC as accession number 141205-05. 
     
     
         21 . A method of reduction of a human pancreatic, ovarian, prostate or colon tumor in a mammal, wherein said human pancreatic, ovarian, prostate or colon tumor expresses at least one epitope of an antigen which specifically binds to the isolated monoclonal antibody produced by the hybridoma deposited with the IDAC as accession number 141205-05 or an antigen binding fragment thereof, which antigen binding fragment is characterized by an ability to competitively inhibit binding of said isolated monoclonal antibody to its target antigen, comprising administering to said mammal said monoclonal antibody or antigen binding fragment thereof in conjunction with at least one chemotherapeutic agent in an amount effective to result in a reduction of said mammal's pancreatic, ovarian, prostate or colon tumor burden. 
     
     
         22 . The method of  claim 21  wherein said isolated monoclonal antibody is conjugated to a cytotoxic moiety. 
     
     
         23 . The method of  claim 22  wherein said cytotoxic moiety is a radioactive isotope. 
     
     
         24 . The method of claim. 21 wherein said isolated monoclonal antibody or antigen binding fragment thereof activates complement. 
     
     
         25 . The method of  claim 21  wherein said isolated monoclonal antibody or antigen binding fragment thereof mediates antibody dependent cellular cytotoxicity. 
     
     
         26 . The method of  claim 21  wherein said isolated monoclonal antibody is a humanized antibody of the isolated monoclonal antibody produced by the hybridoma deposited with the IDAC as accession number 141205-05. 
     
     
         27 . The method of  claim 21  wherein said isolated monoclonal antibody is a chimeric antibody of the isolated monoclonal antibody produced by the hybridoma deposited with the IDAC as accession number 141205-05. 
     
     
         28 . A binding assay to determine a presence of cancerous cells in a tissue sample selected from a human tumor, which is specifically bound by the isolated monoclonal antibody produced by hybridoma cell line AR47A6.4.2 having IDAC Accession No. 141205-05, the humanized antibody of the isolated monoclonal antibody produced by the hybridoma deposited with the IDAC as accession number 141205-05 or the chimeric antibody of the isolated monoclonal antibody produced by the hybridoma deposited with the IDAC as accession number 141205-05, comprising:
 providing a tissue sample from said human tumor;   providing at least one of said isolated monoclonal antibody, said humanized antibody, said chimeric antibody or antigen binding fragment thereof that recognizes the same epitope or epitopes as those recognized by the isolated monoclonal antibody produced by a hybridoma cell line AR47A6.4.2 having IDAC Accession No. 141205-05;   contacting at least one said provided antibodies or antigen binding fragment thereof with said tissue sample; and   determining binding of said at least one provided antibody or antigen binding fragment thereof with said tissue sample;   whereby the presence of said cancerous cells in said tissue sample is indicated.   
     
     
         29 . Use of monoclonal antibodies for reduction of human pancreatic, ovarian, prostate or colon tumor burden, wherein said human pancreatic, ovarian, prostate or colon tumor expresses at least one epitope of an antigen which specifically binds to the isolated monoclonal antibody produced by the hybridoma deposited with the IDAC as accession number 141205-05 or an antigen binding fragment thereof, which antigen binding fragment is characterized by an ability to competitively inhibit binding of said isolated monoclonal antibody to its target antigen, comprising administering to said mammal said monoclonal antibody or antigen binding fragment thereof in an amount effective to result in a reduction of said mammal's human pancreatic, ovarian, prostate or colon tumor burden. 
     
     
         30 . The method of  claim 29  wherein said isolated monoclonal antibody is conjugated to a cytotoxic moiety. 
     
     
         31 . The method of  claim 30  wherein said cytotoxic moiety is a radioactive isotope. 
     
     
         32 . The method of  claim 29  wherein said isolated monoclonal antibody or antigen binding fragment thereof activates complement. 
     
     
         33 . The method of  claim 29  wherein said isolated monoclonal antibody or antigen binding fragment thereof mediates antibody dependent cellular cytotoxicity. 
     
     
         34 . The method of  claim 29  wherein said isolated monoclonal antibody is a humanized antibody of the isolated monoclonal antibody produced by the hybridoma deposited with the IDAC as accession number 141205-05. 
     
     
         35 . The method of  claim 29  wherein said isolated monoclonal antibody is a chimeric antibody of the isolated monoclonal antibody produced by the hybridoma deposited with the IDAC as accession number 141205-05. 
     
     
         36 . Use of monoclonal antibodies for reduction of human pancreatic, ovarian, prostate or colon tumor burden, wherein said human pancreatic, ovarian, prostate or colon tumor expresses at least one epitope of an antigen which specifically binds to the isolated monoclonal antibody produced by the hybridoma deposited with the IDAC as accession number 141205-05 or an antigen binding fragment thereof, which antigen binding fragment is characterized by an ability to competitively inhibit binding of said isolated monoclonal antibody to its target antigen, comprising administering to said mammal said monoclonal antibody or antigen binding fragment thereof; in conjunction with at least one chemotherapeutic agent in an amount effective to result in a reduction of said mammal's human pancreatic, ovarian, prostate or colon tumor burden. 
     
     
         37 . The method of  claim 36  wherein said isolated monoclonal antibody is conjugated to a cytotoxic moiety. 
     
     
         38 . The method of  claim 37  wherein said cytotoxic moiety is a radioactive isotope. 
     
     
         39 . The method of  claim 36  wherein said isolated monoclonal antibody or antigen binding fragment thereof activates complement. 
     
     
         40 . The method of  claim 36  wherein said isolated monoclonal antibody or antigen binding fragment thereof mediates antibody dependent cellular cytotoxicity. 
     
     
         41 . The method of  claim 36  wherein said isolated monoclonal antibody is a humanized antibody of the isolated monoclonal antibody produced by the hybridoma deposited with the IDAC as accession number 141205-05. 
     
     
         42 . The method of  claim 36  wherein said isolated monoclonal antibody is a chimeric antibody of the isolated monoclonal antibody produced by the hybridoma deposited with the IDAC as accession number 141205-05. 
     
     
         43 . A composition effective for treating a human pancreatic, ovarian, prostate or colon tumor comprising in combination:
 an antibody or antigen binding fragment of any one of  claims 1 ,  2 ,  4 ,  5 ,  13 ,  45  or  46 ;   a conjugate of said antibody or an antigen binding fragment thereof with a member selected from the group consisting of cytotoxic moieties, enzymes, radioactive compounds, cytokines, interferons, target or reporter moieties and hematogenous cells; and   a requisite amount of a pharmaceutically acceptable carrier;   wherein said composition is effective for treating said human pancreatic, ovarian, prostate or colon tumor.   
     
     
         44 . An assay kit for detecting the presence of a human cancerous tumor, wherein said human cancerous tumor expresses at least one epitope of an antigen which specifically binds to the isolated monoclonal antibody produced by the hybridoma deposited with the IDAC as accession number 141205-05 or an antigen binding fragment thereof, which antigen binding fragment is characterized by an ability to competitively inhibit binding of said isolated monoclonal antibody to its target antigen, the kit comprising the isolated monoclonal antibody produced by the hybridoma deposited with the IDAC as accession number 141205-05 or an antigen binding fragment thereof, and means for detecting whether the monoclonal antibody, or an antigen binding fragment thereof, is bound to a polypeptide whose presence, at a particular cut-off level, is diagnostic of said presence of said human cancerous tumor. 
     
     
         45 . An isolated monoclonal antibody or antigen binding fragment thereof, which specifically binds to human TROP-2, in which the isolated monoclonal antibody or antigen binding fragment thereof reacts with the same epitope or epitopes of human TROP-2 as the isolated monoclonal antibody produced by a hybridoma cell line AR47A6.4.2 having IDAC Accession No. 141205-05; said isolated monoclonal antibody or antigen binding fragment thereof being characterized by an ability to competitively inhibit binding of said isolated monoclonal antibody to its target human TROP-2 antigen. 
     
     
         46 . An isolated monoclonal antibody or antigen binding fragment thereof that recognizes the same epitope or epitopes as those recognized by the isolated monoclonal antibody produced by the hybridoma cell line AR47A6.4.2 having IDAC Accession No 141205-05; said monoclonal antibody or antigen binding fragment thereof being characterized by an ability to competitively inhibit binding of said isolated monoclonal antibody to its target epitope or epitopes. 
     
     
         47 . A process for reduction of a human pancreatic, ovarian, prostate or colon tumor which expresses at least one epitope of human TROP-2 antigen which is specifically bound by the isolated monoclonal antibody produced by hybridoma cell line AR47A6.4.2 having IDAC Accession No. 141205-05, comprising:
 administering to an individual suffering from said human tumor, at least one isolated monoclonal antibody or antigen binding fragment thereof that binds the same epitope or epitopes as those bound by the isolated monoclonal antibody produced by the hybridoma cell line AR47A6.4.2 having IDAC Accession No. 141205-05;   wherein binding of said epitope or epitopes results in a reduction of pancreatic, ovarian, prostate or colon tumor burden.   
     
     
         48 . A process for reduction of a human pancreatic, ovarian, prostate or colon tumor which expresses at least one epitope of human TROP-2 antigen which is specifically bound by the isolated monoclonal antibody produced by hybridoma cell line AR47A6.4.2 having IDAC Accession No. 141205-05, comprising:
 administering to an individual suffering from said human tumor, at least one isolated monoclonal antibody or antigen binding fragment thereof, that binds the same epitope or epitopes as those bound by the isolated monoclonal antibody produced by the hybridoma cell line AR47A6.4.2 having IDAC Accession No. 141205-05; in conjunction with at least one chemotherapeutic agent;   wherein said administration results in a reduction of tumor burden.   
     
     
         49 . A binding assay to determine the presence of cells which express TROP-2 which is specifically recognized by the isolated monoclonal antibody produced by the hybridoma cell line AR47A6.4.2 having IDAC Accession No. 141205-05, or an antigen binding fragment produced from said isolated monoclonal antibody comprising:
 providing a cell sample;   providing the isolated monoclonal antibody produced by the hybridoma cell line AR47A6.4.2 having IDAC Accession No. 141205-05 or said antigen binding fragment produced from the isolated monoclonal antibody;   contacting said isolated monoclonal antibody or said antigen binding fragment with said cell sample; and   determining binding of said isolated monoclonal antibody or antigen binding fragment thereof with said cell sample;   whereby the presence of cells which express an antigen of TROP-2 which is specifically bound by said isolated monoclonal antibody or said antigen binding fragment is determined.   
     
     
         50 . A method of extending survival and delaying disease progression by treating a human pancreatic, ovarian, prostate or colon tumor in a mammal, wherein said tumor expresses an antigen which specifically binds to the isolated monoclonal antibody produced by the hybridoma cell line AR47A6.4:2 having IDAC Accession No. 141205-05, or an antigen binding fragment produced from said isolated monoclonal antibody comprising administering to said mammal said monoclonal antibody in an amount effective to reduce said mammal's tumor burden, whereby disease progression is delayed and survival is extended. 
     
     
         51 . A method of extending survival and delaying disease progression by treating a human pancreatic, ovarian, prostate or colon tumor in a mammal, wherein said tumor expresses TROP-2 which specifically binds to the isolated monoclonal antibody produced by the hybridoma cell line AR47A6.4.2 having IDAC Accession No. 141205-05, or a TROP-2 binding fragment produced from said isolated monoclonal antibody comprising administering to said mammal said monoclonal antibody in an amount effective to reduce said mammal's tumor burden, whereby disease progression is delayed and survival is extended. 
     
     
         52 . A composition effective for treating a human pancreatic, ovarian, prostate or colon tumor comprising in combination:
 an antibody or antigen binding fragment of any one of  claims 1 ,  2 ,  4 ,  5 ,  13 ,  45  or  46 ; and   a requisite amount of a pharmaceutically acceptable carrier;   wherein said composition is effective for treating said human pancreatic, ovarian, prostate or colon tumor.   
     
     
         53 . A composition effective for treating a human pancreatic, ovarian, prostate or colon tumor comprising in combination:
 a conjugate of an antibody or antigen binding fragment of any one of  claims 1 ,  2 ,  4 ,  5 ,  13 ,  45  or  46  with a member selected from the group consisting of cytotoxic moieties, enzymes, radioactive compounds, cytokines, interferons, target or reporter moieties and hematogenous cells; and   a requisite amount of a pharmaceutically acceptable carrier;   wherein said composition is effective for treating said human pancreatic, ovarian, prostate or colon tumor.