PROTEIN PURIFICATION USING AN AFFINITY MEMBRANE

THE PROPOSED WORK IS DIRECTED TOWARD THE DEVELOPMENT OF A NOVEL AND HIGHLY SELECTIVE MEMBRANE FOR THE CONTINUOUS PURIFICATION OF VALUABLE PROTEINS SUCH AS INTERFERON FROM COMPLEX AND DILUTE MIXTURES. PRODUCTION OF PROTEINS REQUIRES THREE MAIN STEPS: (1) FERMENTATION OR CELL CULTURE;(2) ISOLATION; AND (3) PURIFICATION. ALTHOUGH FERMENTATION AND CELL-CULTURE PROCESSES HAVE UNDERGONE MUCH DEVELOPMENT RECENTLY, ISOLATION AND PURIFICATION REMAIN THE WEAKEST PARTS OF THE OVERALL SCHEME. THE GOAL OF THE PHASE I PROGRAM IS TO ESTABLISH THE FEASIBILITY OF EMPLOYING "AFFINITY MEMBRANES" TO PURIFY HYDROPHOBIC PROTEINS. THE AFFINITY MEMBRANE CONSISTS OF A MICROPOROUS MEMBRANE IN WHICH THE PORES HAVE BEEN FILLED WITH AN AFFINITY LIGAND THAT SELECTIVELY BINDS THE DESIRED PROTEIN. TRANSPORT OF PROTEINS ACROSS THE FILLED PORES OCCURS BY DISSOLUTION INTO AND DIFFUSION THROUGH THE PORE. IN PHASE I OF THIS PROJECT, THE AFFINITY MEMBRANE WILL BE DEVELOPED AND USED TOSELECTIVELY EXTRACT ALPHA-CHYMOTRYPSIN FROM A MIXTURE OF TWOPROTEINS. ALPHA-CHYMOTRYPSIN IS AN INEXPENSIVE PROTEIN THATWILL BE SUITABLE FOR USE IN THE PHASE I FEASIBILITY WORK.