METHOD FOR PURIFICATION OF KOREA TYPE HCV ANTIGEN KHCV UB 897 PROTEIN

摘要

A recombinant strain E. coli W3110 (ATCC 68640) carrying recombinant vector ptrpH-UB-KHCV 897 was cultured in a LB medium containing ampicillin (50 μg/ml) for 12 hrs with shaking. The preculture was inoculated to a M9 medium and cultured for 3-4 hrs. Indoleacrylic acid (final conc. 1.4 mM) was added at OD650 = 0.5 and cultured for 5 hrs further. After recovering of cell pellet by centrifugation, the cell pellet was treated by lysozyme and sonication. Insoluble protein precipitate was isolated by centrifugation and dissolved in a buffer containing 8M urea. The specific antigen KHCV UB 897 was purified by S-Sepharose ion-exchange chromatography and Mono-Q ion-exchange chromatography of FPLC.