Isolation of purified H and L polypeptide chains from guinea-pig gamma-2-immunoglobulin after mild reduction

Abstract

A method is described whereby biologically active H and L chains, free from each other by immunochemical criteria, can be prepared from guinea-pig γ₂-immunoglobulin. The procedure makes use of mild reduction and alkylation in the absence of a denaturing agent, followed by gel filtration in the presence of 4 M guanidine·HCl.

Specific binding of antigen by the method of equilibrium dialysis was found only when specific H chains were mixed with L chains, with more activity resulting when the L chains were also specific. Specific binding by antibody H chains alone was not found.