Screening of hybridoma supernatants raised against membrane fractions from breast cancer using an immunodot assay.

Abstract

Membranes were prepared from mammary tumors and fractionized using gel filtration and gradient polyacrylamide gel electrophoresis. To obtain monoclonal antibodies against tumor-associated antigens, mice were immunized against membrane fraction 2a. After hybridization, we obtained 746 hybridoma cell lines. We performed the immunodot assay for screening. Every supernatant was tested against 15 antigens. Eight antigens were purified fractions from tumor membrane preparation, and five were crude membrane preparations from benign and malignant breast cell lines. Additionally, the Thomsen-Friedenreich antigen of erythrocytes was tested. We selected 83 hybridomas for further characterization.