Assay

Abstract

This disclosure relates to a diagnostic/prognostic assay for prostate cancer and including kits used in said assay.

1 . A diagnostic or prognostic assay for the detection of prostate cancer cells in a biological sample comprising the steps:
 i) obtaining an isolated cell sample from said biological sample and contacting the isolated cell sample with a binding agent that specifically binds at least one MCM polypeptide;   ii) contacting the isolated cell sample with a further binding agent that specifically binds an androgen receptor polypeptide [AR];   optionally iii) contacting the isolated cell sample with a still further binding agent that specifically binds prostate specific antigen [PSA];   iv) detecting the binding of two or more binding agents; and   v) determining the number of cells in said isolated cell sample that positively bind two or more binding agents, wherein the number of positive cells is an indicator of the presence of prostate cancer cells in the biological sample.   
     
     
         2 . An assay according to  claim 1 , wherein said MCM polypeptide is selected from the group consisting of: MCM2, MCM3, MCM4, MCM5, MCM6 or an MCM7 polypeptide. 
     
     
         3 . An assay according to  claim 2  wherein said MCM polypeptide is MCM2 and/or MCM7. 
     
     
         4 . An assay according to  claim 2  or  3  wherein said MCM polypeptide is MCM2. 
     
     
         5 . An assay according to  claim 2  or  3  wherein said MCM polypeptide is MCM7. 
     
     
         6 . An assay according to  claim 3  wherein said MCM polypeptide is MCM2 and MCM7. 
     
     
         7 . An assay according to any of  claims 1 - 6  wherein said MCM polypeptide is represented by the amino acid sequence in  FIG. 1 ,  2 ,  3 ,  4 ,  5  or  6 . 
     
     
         8 . An assay according to any of  claims 1 - 7  wherein said cell sample is contacted with a binding agent that specifically binds AR and a binding agent that specifically binds PSA. 
     
     
         9 . An assay according to  claim 8  wherein said assay detects at least one MCM polypeptide, AR and PSA. 
     
     
         10 . An assay according to  claim 9  wherein said assay detects MCM2 and/or MCM7, AR and PSA. 
     
     
         11 . An assay according to any of  claims 1 - 10  wherein steps (ii) and (iii) are reversed. 
     
     
         12 . An assay according to any of  claims 1 - 11  wherein said binding agent is a monoclonal antibody, or active binding fragment thereof. 
     
     
         13 . An assay according to any of  claims 1 - 12  wherein said biological sample is a sample of isolated bodily fluid. 
     
     
         14 . An assay according to  claim 13  wherein said bodily fluid is urine, semen or seminal fluid. 
     
     
         15 . An assay according to  claim 13  or  14  wherein said biological sample is a filtered wherein said filtered sample provides said isolated cell sample. 
     
     
         16 . A diagnostic or prognostic assay for the determination of prostate cancer in a subject comprising the steps:
 i) obtaining a first isolated biological sample and contacting the sample with a binding agent that specifically binds PSA; optionally   ii) detecting the presence of PSA in said sample;   iii) obtaining a second biological sample from the subject, preparing an isolated cell sample and contacting said cell sample with first and second binding agents that specifically bind at least one MCM polypeptide and an androgen receptor polypeptide;   iv) detecting the presence of said MCM polypeptide[s] and androgen receptor; and   v) determining the number of cells in said isolated cell sample that positively bind both MCM and androgen receptor, wherein the number of positive cells is an indicator of the presence of prostate cancer cells.   
     
     
         17 . An assay according to  claim 16  wherein said first biological sample comprises serum. 
     
     
         18 . An assay according to  claim 16  wherein said second biological sample is urine or semen. 
     
     
         19 . An assay according to any of  claims 16 - 18  wherein said MCM polypeptide is MCM2 and/or MCM7. 
     
     
         20 . An assay according to  claim 19  wherein said MCM polypeptide is MCM2. 
     
     
         21 . An assay according to  claim 19  wherein said MCM polypeptide is MCM7. 
     
     
         22 . An assay according to  claim 19  wherein said MCM polypeptide is MCM2 and MCM7. 
     
     
         23 . An assay according to any of  claims 1 - 22  wherein said detection of binding agents is by fluorescence emission. 
     
     
         24 . An assay according to any of  claims 1 - 22  wherein said detection of binding agents is by enzymic means. 
     
     
         25 . A kit comprising first, second and third binding agents wherein said agents bind a MCM polypeptide, androgen receptor polypeptide and prostate specific antigen polypeptides respectively, 
     
     
         26 . A kit according to  claim 25  wherein said binding agents are monoclonal antibodies.