MEMBRANE-BASED THIOL/DISULFIDE EXCHANGE

PROTEINS ARE INCREASINGLY BEING USED AS PHARMACEUTICALS, DIAGNOSTIC AGENTS, CATALYSTS IN BIOCHEMICAL PROCESSES, AND GROWTH PROMOTERS IN MAMMALIAN CELL CULTURE. COMMERCIAL PROCESSES INVOLVING THE ISOLATION, STORAGE, AND USE OF THIOL-CONTAINING PROTEINS ARE HAMPERED BY THE LACK OF PRACTICAL PROCESSES TO MANIPULATE THE OXIDATION STATE OF PROTEIN THIOLS. ALTHOUGH PROTEIN THIOLS CAN BE MANIPULATED BY THIOL/DISULFIDE EXCHANGE, CONVENTIONAL THIOL/DISULFIDE EXCHANGE REQUIRES ADDING A STOICHIOMETRIC AMOUNT OF EXPENSIVE AND POTENTIALLY HAZARDOUS THIOL REAGENT AND IS RESTRICTED TO ONLY A FEW THIOL REAGENTS. THIS PROJECT WILL DEMONSTRATE THE FEASIBILITY OF A NOVEL METHOD TO CARRY OUT THIOL/DISULFIDE EXCHANGE THAT USES A CATALYTIC AMOUNT OF THIOL REAGENT THAT IS CONTINUOUSLY REGENERATED VIA A MEMBRANE-BASED PROCESS.