Comparative analysis of monoclonal antibody-drug conjugate binding by flow cytometry

摘要: Flow cytometric methods for the evaluation of cell surface binding properties monoclonal antibody (MoAb)-drug/toxin conjugates defining tumor-associated antigens are presented. In these techniques, suspension cultures solid human tumor lines incubated with either varying dilutions MoAb or MoAb-drug followed by FITC-conjugated anti-mouse immunoglobulin antibodies in an indirect assay MoAbs specific target line a competition assay. The amount fluorescent probe bound is measured flow cytometry and mean fluorescence intensity determined. relative capacity quantified linear regression versus concentration primary conjugate. application techniques to several drug toxin KS1/4, which defines adenocarcinoma-associated antigen, demonstrates that assays can be effectively utilized monitor effects covalent chemical modification on MoAb's antigen reactivity.