METHOD RELATING TO SELECTION OF CDNA CLONE ENCODING PROTEIN OF C-TERMINAL REGION OF HYBRIDOMA AND ANTIGEN PRODUCING P-GLYCOPROTEIN CELL SURFACE ANTIGEN SPECIFIC ANTIBODY

摘要

The invention provides a method for selecting hybridomas which produce antibodies specific to domains of a cell surface antigen which is usually not accessible at the surface of intact cells. The method employs the screening of the hybridoma clones obtained for the production of antibodies specific against the cell surface antigen by use of immunoblotting analysis, namely by screening the clones against antigen immobilized on a solid substrate such as nitrocellulose. The invention also includes those hybridomas and monoclonal antibodies when produced according to the method. The method provides monoclonal antibodies to P-glycoprotein surface antigen correlated with multidrug resistance. The antibodies are used to obtain a cDNA probe which in turn was used to select a cDNA clone encoding for a portion of the p-glycoprotein including the C-terminal end. The C-terminal portion of the P-glycoprotein comprising about 239 amino acids and localized to the cytoplasmic side of the plasma membrane contains the epitopes for binding of the monoclonal antibodies.