Detection of breast tumor associated mucin epitope on CAMA cell line using monoclonal antibody G3F1 generated against HMFG membrane.

Abstract

The expression of breast tumor associated mucin epitope on CAMA cell line was detected employing the mouse MAB G3F1 generated against HMFG membrane. Immunocytochemical studies revealed that MAB G3F1 strongly reacted with 85% of breast cancer tissue sections with specific staining of apical cell membrane of malignant epithelial cells. The mucin antigen recognised by MAB G3F1 was detected by selectively extracting high molecular weight glycoprotein antigen from HMFG membrane using lectin affinity chromatography and gel filtration chromatography. Immunoprecipitation studies revealed that MAB G3F1 recognized a high molecular weight glycoprotein with an approximate molecular weight of 300kd. The expression of MAB G3F1 reactive antigen on CAMA cells was detected by immunocytochemistry and by immumoprecipitating 300kd antigen from 125I labelled Tx100 solubilized extract of CAMA cells. The results from these investigations suggest that CAMA cells express MAB G3F1 reactive antigen with tumor associated epitope, similar to tumor associated mucin epitope of HMFG membrane.