Characterization of monoclonal antibodies raised against the prostatic cancer cell line PC‐82

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Abstract
For production of monoclonal antibodies (McAbs), hybrid cells were prepared by fusion of spleen cells of BALB/c mice immunized with the human prostatic cancer cell line PC‐82 and the P3‐X63Ag8. 653 murine myeloma cell line. Supernatants of approximately 500 hybrid clones were screened for prostate specific antibodies using an ELISA on PC‐82 cells. A selection of antibodies was further tested for their specificity on a large series of different tissues. A broad cross reactivity pattern was obtained. Most cross reactivity was with pancreatic tissue, kidney, and bowel. One antibody turned out to react with prostate stromal cells. Two McAbs (ER‐Pr 1 and ER‐Pr 2) reacted solely with prostatic epithelium. Monoclonal antibody affinity chromatography combined with SDS‐PAGE showed that both antibodies were directed against a 35‐kD protein. Immunoblotting revealed that this protein is identical to prostatic antigen (PA). The epitope detected by ER‐Pr 1 and ER‐Pr 2 was largely preserved after formalin‐fixation of prostatic tissues which renders these antibodies very suitable for routine examination of tissue sections.