REVERSED MICELLAR FRACTIONATION OF RECOMBINANT-DNA THERAPETIC PROTEIN INTERMEDIATES

REVERSED MICELLES HOLD MUCH PROMISE AS A MEANS OF CARRYING OUT SELECTIVE LIQUID-LIQUID EXTRACTION OF PROTEINS FROM SOLUTION MIXTURES, SUCH THAT THE DESIRED PROTEIN CAN BE OBTAINED IN PURER AND MORE CONCENTRATED FORM THAN THE ORIGINAL FEED MIXTURE. REVERSED MICELLAR-ORGANIC PHASE (RM-ORG) PARTITIONING COULD HAVE SIGNIFICANT UTILITY IN THE PRIMARY PURIFICATION OR INITIAL FRACTIONATION OF BIOSYNTHETIC PROTEIN PRODUCTION PROCESSES. BIO-ENG IS DEVELOPING A CONTINUOUS FLOW RM-ORG PROCESS FOR THE INITIAL FRACTIONATION OR PURIFICATION OF RECOMBINANT-DNA THERAPEUTICPROTEIN INTERMEDIATES WHICH ARE HARVESTED FROM A FERMENTATION BORTH. THE PROPOSED RM-ORG PROCESS WILL NOT COMPETE WITH, BUT WILL RATHER COMPLEMENT HIGH RESOLUTION CHROMATOGRAPHIC PURIFICATION STEPS. RM-ORG FRACTIONATION WOULD INCREASE THROUGHPUT CAPACITY BY REDUCING THE LOADING ON HIGHER RESOLUTION CHROMATOGRAPHIC PURIFICATION STEPS, ANDWOULD THUS REDUCE PRODUCTION TIME AND COSTS. WE ANTICIPATE MODEST TO HIGH IMPROVEMENTS IN SPECIFICITY (2 TO 10 FOLDE AND HIGH YIELDS (95 TO 100 %). WE ALSO ANTICIPATE PROCESS STREAM CLARIFICATION, CONCENTRATION AND BUFFER EXCHANGE AS INTRINSIC BENEFITS TO THE RM-ORG PROCESS. POTENTIAL ADVANTAGES OF RM-ORG OVER EXISTING PRIMARY PURIFICATION PROCESSES INCLUDE SELECTIVITY, SPEED OF PROCESSING, PRESERVATION OF PROTEIN INTEGRITY, AND SCALABILITY.