Conformation dependence of a monoclonal antibody defined epitope on free human kappa chains.

Chemically and enzymatically modified kappa chains were tested by inhibition radioimmunoassay for their ability to block the binding of antibody K-1-21 with native kappa chains. Complete reduction and carboxymethylation of intrachain disulphide bonds destroyed the free kappa-chain epitope, a result confirmed by Western blotting of unreduced and reduced kappa monomers and dimers. Purified V kappa fragments failed to block the homologous interaction while inhibition was obtained with a pepsin digest yielding predominantly the C kappa region. Dimeric kappa chains were less effective than monomers in the inhibition assay, although HPLC analysis of immune complexes demonstrated the binding of two antibody molecules per molecule of dimer. Thus, the epitope on free kappa chains recognized by K-1-21 is dependent upon conformational integrity of the C kappa domain, the decreased binding activity of dimeric chains possibly being due to minor conformational changes induced by C-domain interactions.