Rittenhouse J, Turon M C, Helfrich R J, Albrecht K S, Weigl D, Simmer R L, Mordini F, Erickson J, Kohlbrenner W E
Pharmaceutical Products Division, Abbott Laboratories, IL 60064-3500.
Biochem Biophys Res Commun. 1990 Aug 31;171(1):60-6. doi: 10.1016/0006-291x(90)91356-w.
A procedure is described which employs pepstatin-agarose for the affinity purification of either HIV-1 or HIV-2 protease from two similar recombinant E. coli constructs that were developed for the expression of these enzymes. HIV-2 protease was routinely expressed at much higher levels than the HIV-1 enzyme and pepstatin-agarose was the only chromatography step required to isolate pure HIV-2 protease from crude bacterial lysates. A Mono S ionic exchange step following pepstatin-agarose chromatography was sufficient to bring the HIV-1 protease to homogeneity. Purification of either enzyme can be completed in several days yielding homogeneous preparations suitable for crystallization and other physical characterization.
