PROCEDURE FOR REFOLDING AND PURIFICATION OF RECOMBINANT PROTEINS FROMESCHERICHIA-COLI INCLUSION-BODIES USING A STRONG ANION-EXCHANGER

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.