AFFINITY MATRICES FOR PROCESS SCALE PURIFICATION

THE AIM OF THIS RESEARCH IS TO INVESTIGATE THE USE OF NEW AFFINITY MATRICES IN PURIFICATION AND CLEAVAGE OF RECOMBINANT PROTEINS AT PROCESS SCALE. THE PERFORMANCE OF DIFFERENT AFFINITY MATRICES FOR IMMOBILIZATION OF ENZYMESWILL BE DIRECTLY COMPARED. SCALABILITY AND PERFORMANCE WILLBE EVALUATED IN RELATION TO COST. IN PARTICULAR, THE PERFORMANCE OF MEMBRANE-IMMOBILIZED ENZYME WILL BE COMPARED TO SOFT-GEL AND RIGID-BEAD IMMOBILIZED ENZYMES. THE RESULTS COULD AID IN THE DESIGN OF AFFINITY MATRICES USEFUL FOR PROCESS SCALE WORK. BY STREAMLINING PURIFICATIONPROCESSES SIGNIFICANTLY, APPLICABLE AFFINITY METHODS COULD FACILITATE THE DEVELOPMENT OF NEW PROTEIN THERAPEUTICS. A SMALL DOMAIN THAT CAN BE PURIFIED BY A SCALABLE AFFINITY CHROMATOGRAPHY METHOD WILL BE DEFINED BY PEPTIDE SYNTHESIS. PEPTIDE SEQUENCES TAKEN FROM CALCIUM-BINDING DOMAINS WILL BE SYNTHESIZED. AFTER INITIAL SCREENING TO IDENTIFY PEPTIDES HAVING AFFINITY FOR CA(2+), AN AFFINITY PURIFICATION METHOD BASED UPON METAL BINDING WILL BE OPTIMIZED. THE ULTIMATE GOAL IS TO EXPRESS THE SHORTEST ACTIVE SEQUENCE RECOMBINANTLY IN FUSION WITH POTENTIAL PRODUCT PROTEIN SEQUENCES.