A NEW LARGE-SCALE AFFINITY SEPARATION TECHNIQUE

THE PROPOSED RESEARCH IS DIRECTED AT DEVELOPING A NEW AFFINITY ADSORPTION AND CHROMATOGRAPHY TECHNIQUE WHICH OFFERS GREAT PROMISE FOR COMMERCIAL SCALE SEPARATIONS OF BIOLOGICALLY ACTIVE MATERIALS SUCH AS INTERFERON AND GROWTH HORMONE. THE TECHNIQUE IS BASED ON ATTACHING THE AFFINITY LIGANDS TO A HYDROPHOBIC LIQUID INTERFACE RATHER THAN TO A SOLID SUPPORT AND IS DESIGNATED LIQUID-LIQUID AFFINITY ADSORPTION AND CHROMATOGRAPHY. IN PHASE I THE FEASIBILITY AND POTENTIAL OF EMULSION AFFINITY ADSORPTION AND OF EMULSION AFFINITY CHROMATOGRAPHY WILL BE DEMONSTRATED USING A FLUOROCARBON AS THE HYDROPHOBIC LIQUID AND USING BOVINE PANCREATIC ALPHA CHYMOTRYPSIN ENZYME AS THE MODEL SYSTEM. THE PURIFICATION AND RECOVERY ACHIEVABLE IN SINGLE STAGE EMULSION AFFINITY ADSORPTION AND PURITY, RECOVERY AND THROUGHPUT ACHIEVABLE WITH EMULSION AFFINITY CHROMATOGRAPHY WILL BE DETERMINED. THE RESULTS WILL INDICATE THE RELATIVE EMPHASIS WHICH SHOULD BE GIVEN THESE TWO APPROACHES IN PHASE II. KEY BENEFITS OF THE NEW TECHNIQUE RESULT FROM ITS SUITABILITY FOR CONTINUOUS COUNTERCURRENT OPERATION, FROM THE SIMPLIFICATION OF PRODUCT RECOVERY RESULTING FROM THE ABILITY TO COALESCE THE PRODUCT EMULSION AND ISOLATE THE INTERFACIAL MATERIAL INCLUDING THE AFFINITY LIGAND-PRODUCT COMPLEX WITH MINIMAL WASHING AND SIMPLIFIED ELUTION, AND FROM THE REDUCTION OF STERIC EFFECTS.