J. Suttnar et al., PROCEDURE FOR REFOLDING AND PURIFICATION OF RECOMBINANT PROTEINS FROMESCHERICHIA-COLI INCLUSION-BODIES USING A STRONG ANION-EXCHANGER, Journal of chromatography B. Biomedical applications, 656(1), 1994, pp. 123-126
Citations number
12
Categorie Soggetti
Chemistry Analytical
Journal title
Journal of chromatography B. Biomedical applications
Using Escherichia coli system expressing papilloma virus HPV16 E7MS2 f
usion protein as a model system, a novel procedure was applied to solu
bilize, purify and refold recombinant proteins from E. coli inclusion
bodies. The necessity to reactivate proteins at low protein concentrat
ions (owing to their tendency to aggregate at high concentrations) was
overcome by solubilization of inclusion bodies in alkaline solution a
nd immobilization of proteins on a strong and resistant anion exchange
r. This procedure has an inherent advantage of combining refolding and
purification procedures in one step. The solubilization of the fusion
protein in an alkaline reagent with the use of an anion exchanger res
ulted in considerable purification of the recombinant protein at a fai
rly high concentration. The protein was soluble under mild conditions
and reacted with antibodies against the ''native'' papilloma virus.