PURIFICATION METHOD OF RECOMBINANT STREPTOKINASE EXPRESSED AS INCLUSION BODY

摘要

PURPOSE: A purification method of recombinant streptokinase expressed as an inclusion body is provided, thereby purifying a large quantity of recombinant streptokinase having high activity in higher yield. CONSTITUTION: A purification method of recombinant streptokinase expressed from transformed E. coli as an inclusion body comprises the steps of: (a) washing a transformed E. coli with a buffer solution having pH 4.5 to 5.5 and pulverizing the E. coli to recover protein precipitates; (b) dissolving the protein precipitates using amphiphilic salt containing solution and restoring the activity of the protein using a neutral buffer solution having higher ionic strength; and (c) subjecting the restored protein to a column chromatography, wherein the buffer solution of the step (a) is 20mM to 50mM of sodium citric acid buffer solution; the amphiphilic salt containing solution of the step (b) is 6M to 10M of urea; the neutral buffer solution having higher ionic strength of the step (b) is 20mM to 100mM of sodium chloride; and the column chromatography of the step (c) is anion exchange column chromatography and gel filtration column chromatography.