IMPROVED METHOD FOR THE REFOLDING OF PROTEINS

摘要

A novel, generally applicable method for producing correctly folded proteins from a mixture of misfolded proteins, e.g. bacterial inclusion-body aggregates. A major new aspect of the method is that over-al l efficiency is achieved by subjecting proteins to a tim e- sequence of multiple denaturation-renaturation cycles, resulting in gradual accumulation of the correctly folded protein. The metho d has proven efficient for a variety of recombinant proteins. Also provided are n ovel encrypted recognition sites for bovine coagulation factor Xa. The encrypted recognition sites described may be activated in vitro by c ontrolled oxidation or by reversible derivatization of c ysteine residues and thereby generate new cleavage sites for factor Xa. Two new reco mbinant serine protease exhibiting narrow substrate spec ificity for factor Xa recognition sites are also provided. They may replace natural coagulation factor Xa for cleavage of chimeric proteins .