PURIFICATION OF HUMAN INTERLEUKIN-4 FROM A SECRETED ESCHERICHIA COLI MUTANT.

摘要

ACTIVE RECOMBINANT HUMAN IL-4 IS PURIFIED FROM A CRUDE FERMENTATION BROTH OF A SECRETED E.COLI MUTANT BY SUBJECTING SAID SOLUTION TO SELECTIVE CHROMATOGRPHY ON A ZINC-CHELATING AGAROSE COLUMN AT pH 7.2 IN A BUFFER CONTAINING A HIGH CONCENTRATION OF SODIUM CHLORIDE, I.E., ABOUT 1 M SODIUM CHLORIDE, WASHING FIRST WITH A PHOSPHATE BUFFER AT, pH 7.2 CONTAINING 1.0M SODIUM CHLORIDE THEN WITH A BUFFER CONTAINING 10% GLYCEROL AND ELUTING AT pH 5.0. THE RESULTING PURIFIED ACTIVE RECOMBINANT HUMAN IL-4 SOLUTION IS FURTHER TREATED BY CATION EXCHANGE CHROMATOGRPHY,CONCENTRATION BY DIAFILTRATION UP TO 20 MG/ML AND SIZE EXCLUSION GEL CHROMATOGRAPHY USING A CITRATE BUFFER AT pH 4.5.