US2008206140A1PendingUtilityA1

Cspcna Isoform Antibodies and Uses Thereof

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Assignee: UNIV INDIANA RES & TECH CORPPriority: Apr 27, 2005Filed: Apr 27, 2006Published: Aug 28, 2008
Est. expiryApr 27, 2025(expired)· nominal 20-yr term from priority
Inventors:Robert J. HickeyLinda H. MalkasLauren Schnaper
C07K 2317/76A61P 35/00C07K 16/44A61K 2039/505C07K 2317/34C07K 16/3015A61P 43/00G01N 2333/47A61K 45/06A61K 9/127C07K 2317/73A61K 39/39558C07K 16/18G01N 33/57595
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Claims

Abstract

Antibodies specifically bind only to a cancer specific proliferating cell nuclear antigen (csPCNA) isoform and not to the non-malignant proliferating cell nuclear antigen (nmPCNA) isoform. Methods and compositions to detect the presence of csPCNA isoform are disclosed.

Claims

exact text as granted — not AI-modified
1 . An isolated antibody that specifically binds cancer specific proliferating cell nuclear antigen (csPCNA) isoform. 
     
     
         2 . The antibody of  claim 1 , wherein the csPCNA isoform comprises an amino acid sequence of SEQ ID NO: 3. 
     
     
         3 . The antibody of  claim 1 , wherein the antibody binds to an epitope comprising an amino acid sequence within the csPCNA protein that binds to Xeroderma pigmentosum group G (XPG) protein. 
     
     
         4 . The antibody of  claim 1 , wherein the antibody binds to an epitope of csPCNA comprising an amino acid sequence selected from LGIPEQEY (SEQ ID NO: 1), VEQLGIPEQEY (SEQ ID NO: 5), LGIPEQEYSCVVK (SEQ ID NO: 6), LGIPEQEYSCVVKMPSG (SEQ ID NO: 7), EQLGIPEQEY (SEQ ID NO: 8), QLGIPEQEY (SEQ ID NO: 9), LGIPEQEYSCVVKMPS (SEQ ID NO: 10), LGIPEQEYSCVVKMP (SEQ ID NO: 11), LGIPEQEYSCVVKM (SEQ ID NO: 12), LGIPEQEYSCVV (SEQ ID NO: 13), LGIPEQEYSCV (SEQ ID NO: 14), and LGIPEQEYSC (SEQ ID NO: 15). 
     
     
         5 . The antibody of  claim 1  is a monoclonal antibody. 
     
     
         6 . The antibody of  claim 1  is a chimeric antibody. 
     
     
         7 . The antibody of  claim 1  is a recombinant antibody. 
     
     
         8 . The antibody of  claim 1  is a single chain antibody. 
     
     
         9 . The antibody of  claim 1 , wherein the antibody is an antibody fragment selected from Fab, Fab′, or F(ab′) 2 . 
     
     
         10 . The antibody of  claim 1 , wherein the antibody is associated with a detectable agent. 
     
     
         11 . The antibody of  claim 10 , wherein the detectable agent is selected from a fluorescent label, radio label, chromatogenic label, and an enzymatic label. 
     
     
         12 . A composition comprising an isolated and substantially purified antibody that is specifically bound to an epitope of cancer specific proliferating cell nuclear antigen (csPCNA), wherein the epitope comprises an amino acid sequence of LeuGlyIleProGluGlnGluTyr (SEQ ID NO: 1). 
     
     
         13 . Use of a csPCNA specific antibody to detect a cancer specific proliferating cell nuclear antigen (csPCNA) isoform in a biological sample comprising:
 contacting the biological sample with the antibody of  claim 1  that specifically binds cancer specific proliferating cell nuclear antigen (csPCNA) isoform;   providing conditions for the antibody binding; and   detecting the binding of the antibody with the csPCNA isoform.   
     
     
         14 . The use of  claim 13 , wherein the biological sample is a bodily fluid. 
     
     
         15 . The use of  claim 14 , wherein the bodily fluid is selected from blood, plasma, lymph, serum, pleural fluid, spinal fluid, saliva, sputum, urine, gastric juice, pancreatic juice, ascites fluid, synovial fluid, milk, and semen. 
     
     
         16 . The use of  claim 13 , wherein the biological sample is a tissue sample. 
     
     
         17 . The use of  claim 16 , wherein the tissue is selected from breast, prostrate, lung, colon, epithelial, connective, cervical, esophageal, brain, thymus, thyroid, pancreas, testis, ovary, intestine, bladder, stomach, soft tissue sarcomas, osteosarcoma, leukemia, lymphoma, carcinoma, adenocarcinoma, placenta, fibrous, germ cell tissue, and extracts thereof. 
     
     
         18 . The use of  claim 13 , wherein the antibody detection is performed in vivo. 
     
     
         19 . The use of  claim 13 , wherein the antibody detection is performed by providing a labeled secondary antibody. 
     
     
         20 . The use of  claim 13 , wherein the antibody that specifically binds cancer specific proliferating cell nuclear antigen (csPCNA) isoform is labeled. 
     
     
         21 . The use of  claim 13 , wherein the detection of csPCNA isoform is performed using a mass spectrometric analysis. 
     
     
         22 . The use of  claim 13 , wherein the detection of csPCNA isoform is performed using an enzyme linked immunosorbent assay. 
     
     
         23 . The use of  claim 13 , wherein the detection of csPCNA isoform is performed using an immunohistochemical method. 
     
     
         24 . Use of a csPCNA specific antibody to diagnose or prognose malignancy comprising the steps of:
 detecting csPCNA in a biological sample by an antibody that specifically binds cancer specific proliferating cell nuclear antigen (csPCNA) isoform; and   diagnosing malignancy based on the detection of csPCNA in the biological sample.   
     
     
         25 . The use of  claim 24 , wherein the biological sample is a biological tissue or a fluid. 
     
     
         26 . A method for producing antibodies specific to a cancer specific proliferating cell nuclear antigen (csPCNA) isoform comprising:
 administering to an antibody generation source an immunogenic amount of a peptide representing an epitope specific to csPCNA isoform, wherein the peptide is selected from contiguous or non-contiguous amino acid residues on the region of csPCNA that interacts with a Xeroderma pigmentosum group G (XPG) protein;   providing conditions for antibody generation; and   isolating and purifying the antibodies.   
     
     
         27 . The method of  claim 26 , wherein the antibodies are isolated and purified from hybridoma cells. 
     
     
         28 . The method of  claim 26 , wherein the peptide comprises an amino acid sequence selected from LGIPEQEY (SEQ ID NO: 1), VEQLGIPEQEY (SEQ ID NO: 5), LGIPEQEYSCVVK (SEQ ID NO: 6), LGIPEQEYSCVVKMPSG (SEQ ID NO: 7), EQLGIPEQEY (SEQ ID NO: 8), QLGIPEQEY (SEQ ID NO: 9), LGIPEQEYSCVVKMPS (SEQ ID NO: 10), LGIPEQEYSCVVKMP (SEQ ID NO: 11), LGIPEQEYSCVVKM (SEQ ID NO: 12), LGIPEQEYSCVV (SEQ ID NO: 13), LGIPEQEYSCV (SEQ ID NO: 14), and LGIPEQEYSC (SEQ ID NO: 15). 
     
     
         29 . The method of  claim 26 , wherein the peptide comprises the amino acid sequence of CGGGLGIPEQEY (SEQ ID NO: 2). 
     
     
         30 . The method of  claim 26 , wherein the peptide is associated with a carrier protein. 
     
     
         31 . The method of  claim 30 , wherein the carrier protein is keyhole limpet hemocyanin (KLH). 
     
     
         32 . Use of a csPCNA specific antibody to identify the location of a tumor in vivo, the method comprising:
 administering a cancer specific proliferating cell nuclear antigen (csPCNA) isoform specific antibody of  claim 1  that specifically binds csPCNA, wherein the antibody is labeled with a detectable agent; and   determining the location of the tumor by detecting the accumulation of the labeled csPCNA-specific antibody at the tumor site.   
     
     
         33 . Use of a csPCNA specific antibody to augment reduction of tumor progression comprising:
 providing a pharmaceutically acceptable composition comprising a formulation of a therapeutically effective amount of cancer specific proliferating cell nuclear antigen (csPCNA) isoform-specific antibody of  claim 1  and a delivery component;   administering the formulation; and   reducing the progression of tumor by delivering the formulation comprising csPCNA-specific antibody to the tumor site, wherein the csPCNA-specific antibody reacts with csPCNA isoform present in tumor cells.   
     
     
         34 . The use of  claim 33 , wherein the formulation comprises a liposome or a nanoparticle. 
     
     
         35 . The method of  claim 33 , wherein the formulation comprises a tumor killing agent or an immune enhancing agent. 
     
     
         36 . Use of a csPCNA specific antibody to identify an anti-cancer agent comprising:
 contacting a population of cancer cells with an agent;   measuring the levels of a cancer specific proliferating cell nuclear antigen (csPCNA) isoform by assaying the binding of a csPCNA-specific antibody of  claim 1  to the csPCNA isoform; and   determining that the agent is an anticancer agent if the levels of csPCNA isoform in the cancer cells contacted with the agent are less than the levels of csPCNA isoform in cancer cells not contacted with the agent.   
     
     
         37 . The use of  claim 36 , wherein the agent is a small molecule. 
     
     
         38 . The use of  claim 36 , wherein the population of cancer cells is selected from a cancer cell line, xenograft and an orthotopic model system of cancer. 
     
     
         39 . The use of  claim 36 , wherein the determining step comprises measuring the levels of non-malignant PCNA isoform in normal cells contacted with the agent and in normal cells not contacted with the agent. 
     
     
         40 . The use of  claim 36 , wherein the identification of the anti-cancer agent is performed in a high-throughput system. 
     
     
         41 . An immunoassay kit for detecting the csPCNA isoform of PCNA comprising:
 an antibody preparation of  claim 1  that specifically binds only to a cancer specific proliferating cell nuclear antigen (csPCNA) isoform and not to the normal proliferating cell nuclear antigen (nmPCNA) isoform, whereby the antibodies and csPCNA form a complex;   and reagents for detecting the complex.   
     
     
         42 . The immunoassay kit of  claim 41  comprising a peptide of amino acid sequence selected from LGIPEQEY (SEQ ID NO: 1), VEQLGIPEQEY (SEQ ID NO: 5), LGIPEQEYSCVVK (SEQ ID NO: 6), LGIPEQEYSCVVKMPSG (SEQ ID NO: 7), EQLGIPEQEY (SEQ ID NO: 8), QLGIPEQEY (SEQ ID NO: 9), LGIPEQEYSCVVKMPS (SEQ ID NO: 10), LGIPEQEYSCVVKMP (SEQ ID NO: 11), LGIPEQEYSCVVKM (SEQ ID NO: 12), LGIPEQEYSCVV (SEQ ID NO: 13), LGIPEQEYSCV (SEQ ID NO: 14), and LGIPEQEYSC (SEQ ID NO: 15), wherein the peptide is used as a positive control. 
     
     
         43 . The immunoassay kit of  claim 41  comprising a preparation of csPCNA isoform comprising an amino acid sequence of SEQ ID NO: 3, wherein the csPCNA isoform is used as a positive control. 
     
     
         44 . An isolated auto-antibody specific to a cancer specific proliferating cell nuclear antigen (csPCNA) isoform. 
     
     
         45 . The auto-antibody of  claim 44 , wherein the auto-antibody is complexed to an epitope of csPCNA isoform. 
     
     
         46 . A method of determining the presence of malignant cells, the method comprising:
 contacting a biological sample suspected of containing auto-antibodies, to a substrate comprising bound cancer specific proliferating cell nuclear antigen (csPCNA) isoform or fragments thereof, wherein the auto-antibodies are specific to a csPCNA isoform;   providing conditions for csPCNA-auto-antibody complex formation; and   detecting the presence of the auto-antibody-csPCNA complex in the biological sample.   
     
     
         47 . The method of  claim 46 , wherein the presence of the auto-antibody-csPCNA complex is detected using an anti-human secondary antibody. 
     
     
         48 . The method of  claim 46 , wherein the presence of the auto-antibody-csPCNA complex is detected using a labeled biological sample. 
     
     
         49 . The method of  claim 46 , wherein the biological sample is selected from blood, plasma, lymph, serum, pleural fluid, spinal fluid, saliva, sputum, urine, gastric juice, pancreatic juice, ascites fluid, synovial fluid, milk, and semen. 
     
     
         50 . A method of detecting the presence of a circulating cancer specific proliferating cell nuclear antigen (csPCNA) isoform, the method comprising detecting an auto-antibody specific to the csPCNA isoform in a biological sample and thereby determining the presence of circulating csPCNA isoform. 
     
     
         51 . A method of monitoring the remission status of an individual, the method comprising:
 detecting the presence of proliferating cell nuclear antigen (csPCNA) isoform in the individual prior to and after cancer therapy; and   determining the remission status of the individual by comparing the levels of circulating csPCNA isoform prior to and after cancer therapy.   
     
     
         52 . The method of  claim 51 , wherein the csPCNA isoform is detected by determining the presence of auto-antibodies to csPCNA isoform. 
     
     
         53 . The method of  claim 51 , wherein the csPCNA isoform is detected by an antibody specific to the csPCNA isoform. 
     
     
         54 . The method of  claim 51 , wherein the csPCNA isoform is detected by an enzyme linked immunosorbent assay.

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