Kinetics of Divalent Monoclonal Antibody Binding to Tumour Cell Surface Antigens Using Flow Cytometry: Standardization and Mathematical Analysis

Flow cytofluorimetric methods have been used to quantitate the interaction between a divalent monoclonal antibody and a tumour cell surface antigen. After standardization using fluorescein and 125I-labelled antibodies, kinetics of association and dissociation were measured, and antibody bound at equilibrium quantitated. A mathematical model was developed in conjunction with these experimental results which allowed the calculation of rates for monovalent association and monovalent and divalent dissociation, and a description of the contribution of each to the level of bound antibody at different antibody concns.