Investigation of monoclonal anti-glycophorin antibodies pH and temperature dependance, reactivity with enzyme treated cells and flow cytometry studies

Monoclonal antibodies against antigens of the M/N blood group system have been shown to be dependent on the microenvironment concerning their specificity [1, 3]. Therefore all the workshop antibodies were first tested at different pH and temperatures to investigate their binding specificity. Concerning the binding specificity another important aspect is the reactivity with enzyme treated red blood cells using trypsin, chymotrypsin and neuraminidase [2]. These tests were performed with all antibodies, which were positive in the direct agglutination tests.

An interesting evaluation for epitope specificity is the analysis of reactivity including inhibition studies using flow cytometry [4]. Binding studies were performed with all workshop antibodies with M- and N-RBC whereas inhibition studies were done only with monoclonals showing anti-M specificity.