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-all efficiency is achieved by subjecting proteins to a time-sequence of multiple denaturation-renaturation cycles, resulting in gradual accumulation of the correctly folded protein. The method has proven efficient for a variety of recombinant proteins. Also provided are novel encrypted recognition sites for bovine coagulation factor Xa. The encrypted recognition sites described may be activated in vitro by controlled oxidation or by reversible derivatization of cysteine residues and thereby generate new cleavage sites for factor Xa. Two new recombinant serine protease exhibiting narrow substrate specificity for factor Xa recognition sites are also provided. They may replace natural coagulation factor Xa for cleavage of chimeric proteins.
申请公布号
WO9418227(A2)
申请公布日期
1994.08.18
申请号
WO1994DK00054
申请日期
1994.02.04
申请人
DENZYME APS;THOGERSEN, HANS, CHRISTIAN;HOLTET, THOR, LAS;ETZERODT, MICHAEL
发明人
THOGERSEN, HANS, CHRISTIAN;HOLTET, THOR, LAS;ETZERODT, MICHAEL