ANALYSIS OF THE INTERACTION BETWEEN A SYNTHETIC PEPTIDE OF INFLUENZA-VIRUS HEMAGGLUTININ AND MONOCLONAL-ANTIBODIES USING AN OPTICAL BIOSENSOR

The interaction between two monoclonal antibodies and their correspond ing Fab' fragments with a synthetic peptide, corresponding to the C-te rminal 23 residues of the HA(1) chain of influenza virus hemagglutinin against which they were generated, has been examined using an optical biosensor employing the detection principal of surface plasmon resona nce (Pharmacia BIAcore(TM)). The data obtained has been analysed in de tail by linear transformation of the primary data and nonlinear regres sion analysis, as well as by analysis of equilibrium binding data. The 2/1 antibodies and their Fab' fragments displayed higher affinity tha n the corresponding 1/1 proteins. The IgGs were found to have equilibr ium association constants (K-A) 10-20-fold higher than the correspondi ng Fab' fragments. This appears largely to be due to differences in th e dissociation rate constant (k(d)) and probably reflects increased av idity due to bivalent binding. Copyright (C) 1996 Elsevier Science Ltd .