NOVEL PROCESS FOR SOLUBILIZING INCLUSION BODIES

A SIGNIFICANT DEVELOPMENT IN RECOMBINANT DNA TECHNOLOGY IS THE CAPABILITY TO EXPRESS EUKARYOTIC GENES IN PROKARYOTES SUCH AS ESCHERICHIA COLI. SINCE THE NATURAL SUPPLY OF MANY EUKARYOTIC PROTEINS OF POTENTIAL CLINICAL OR INDUSTRIAL IMPORTANCE IS LIMITED, GENE CLONING AND EXPRESSION OFFER A POTENTIALLY ABUNDANT SOURCE OF EUKARYOTICPROTEINS. RECOMBINANT PROTEINS EXPRESSED IN PROKARYOTES ARE SEQUESTERED IN THE CYTOPLASM AS INSOLUBLE AGGREGATES KNOWN AS INCLUSION BODIES. CURRENTLY, THE RECOVERY OF RECOMBINANTPROTEINS FROM INCLUSION BODIES REQUIRES THE USE AGGRESSIVE PROTEIN DENATURING AGENTS THAT CAN IRREVERSIBLY DAMAGE THE PROTEIN AND YIELDS VERY LOW PERCENTAGES OF ACTIVE PROTEIN. IN THIS PROPOSAL, WE DESCRIBE A NOVEL PROCESS FOR SOLUBILIZING AND RECOVERING EUKARYOTIC PROTEINS FROM INCLUSION BODIES. THE PROCESS PROMISES TO REPLACE OR ENHANCE THE SOLUBILIZATION AND RECOVERY PROCESS CURRENTLY IN USE. THE PRINCIPAL EXPECTED BENEFIT IS AN IMPROVEMENT IN THE YIELD OF PROTEIN IN ITS ACTIVE FORM.