Biochem Biophys Res Commun. 1990 Aug 31;171(1):60-6.

Affinity purification of HIV-1 and HIV-2 proteases from recombinant E. coli strains using pepstatin-agarose.

Rittenhouse J, Turon MC, Helfrich RJ, Albrecht KS, Weigl D, Simmer RL, Mordini F, Erickson J, Kohlbrenner WE.

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Pharmaceutical Products Division, Abbott Laboratories, IL 60064-3500.

Abstract

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.

PMID:: 2203350; [PubMed - indexed for MEDLINE]

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Affinity purification of HIV-1 and HIV-2 proteases from recombinant E. coli strains using pepstatin-agarose.

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