Preparation of antibody (IgG)-ricin A-chain conjugate and its biologic activity

As a model experiment for the preparation of cancer chemotherapeutic agents, a disulfide-linked conjugate of rabbit anti-mouse IgG (RaMIgG) and ricin A-cahin was synthesized. Activated sulfhydryl groups were first introduced into RaMIgG and then the modified RaMIgG was reacted with ricin A-chain. The fraction containing the conjugate was obtained from the reaction mixture by gel filtration through Sephadex G-150 and was characterized by sodium dodecyl sulfate-polyacrylamide disc gel electrophoresis and double immunodiffusion in agar. The inhibitory activity of A-chain against protein synthesis in a cell-free system and the binding activity of RaMIgG to mouse B lymphocytes were both fully retained. The purified conjugate selectively inhibited protein synthesis in mouse B lymphocytes possessing cell surface Ig, but not that in thymocytes. The cytotoxicity was inhibited by adding mouse IgG and an excess amount of RaMIgG to the cell culture. The effects of unlinked RaMIgG and A-chain on the cytotoxicity were negligible even when the two were used as a mixture.