5E10: a prostate-specific surface-reactive monoclonal antibody

We created mAb that reacted solely with prostate epithelial cells. The strategy involved immunizing mice with a mixture of six different prostatic carcinoma cell lines and selecting by flow cytometry only those antibodies that bind whole cells. The primary screening was performed using a mixture of all six prostate cell lines used in immunization together with six non-prostate cell lines in the same tube. Antibodies that gave a bimodal pattern of surface staining were selected for further evaluation. The most attractive clone, designated 5E10, produced IgG1 mAb that recognized four of the six prostatic cell lines and did not react with non-prostate tumor cell lines, peripheral blood and bone marrow mononuclear cells and endothelial and bone marrow stromal cells. 5E10 mAb reacted with both benign and malignant prostate in eight of eight histological samples and no reactivity was noted with non-prostate normal tissues. The 5E10 antigen is a transmembrane glycoprotein with a molecular weight of 110 kDa and the epitope recognized by 5E10 is extracellular. Ongoing studies are exploring the nature of this antigen in more depth.