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Development of Two New Monoclonal Antibodies Reactive to a Surface Antigen Present on Human Ovarian Epithelial Cancer Cells¹

Department of Pharmacology [F-J. X., Y-H. Y., B-Y. L., C. L., S. R.] and Women's Cancer Center [M. M., S. E., L. C., S. R.], University of Minnesota, Minneapolis, Minnesota 55455

Monoclonal antibodies which bind selectively to cancer cells are currently used for tumor localization and for targeting cytotoxic reagents. The success of these approaches depends on the specificity of the antibody and its reactivity to a majority of the tumor samples. Frequently, monoclonal antibodies are generated by immunizing mice with antigenic preparations from a single tumor cell line. Antibodies generated under these conditions often react to a narrow range of tumors. In the present study, mice were immunized with multiple ovarian cancer cell lines in a sequential manner to amplify the immune response against common antigenic determinants expressed in these cell lines. Spleen cells from the immunized mice were then fused with NS-1 myeloma cells to establish hybridomas. Two cell lines were selected on the basis of their selective reactivity to ovarian cancer cells after extensive screening. Monoclonal antibodies OVX1 and OVX2 bound to all 5 ovarian carcinoma cell lines tested and did not bind to normal fibroblast cells. These antibodies recognized a unique antigenic determinant present in ovarian and breast cancer cells. Cross-blocking studies showed that the binding of OVX1 and OVX2 is not displaceable by 10 other previously described anti-ovarian antibodies including OC125. In immunocytochemical studies, OVX1 reacted to a majority of ovarian cancer tissues (17 of 20) and did not bind to normal ovarian tissues. Preliminary results indicate that OVX1 and OVX2 antibodies are directed to a high molecular weight antigen. These antibodies could be used in the preparation of cytotoxic conjugates.

¹ This work was supported by a grant from the National Cancer Institute, CA 48608 (to S. R.) and a grant from the Cancer Research Foundation of America (to M. M.).

² To whom requests for reprints should be addressed, at 3-249 Millard Hall, Department of Pharmacology, 435 Delaware Street S.E., University of Minnesota, Minneapolis, MN 55455.

Received 1/25/91. Accepted 5/15/91.

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