RENATURATION OF RECOMBINANT PLATELET-DERIVED GROWTH FACTOR (PDGF); MIXED DISULPHIDE COMPOUND OR PDGF; PURE PDGF MONOMER; AND MIXED DIMERIC RECOMBINANT PDGF

摘要

A method for refolding recombinant platelet-derived growth factor from a high expression host cell system, such as E. coli, is provided in accordance with the present invention. The recombinant platelet-derived growth factor is isolated from inclusion body proteins, after which the free sulfydryl groups of the reduced monomeric recombinant protein are blocked. The blocked recombinant protein can then be brought into a biologically active conformation, without interference from the formation of incorrect disulfide bonds. Once in a biologically active conformation, the recombinant platelet-derived growth factor may be unblocked to allow the desired disulfide bonds to form, thus locking the recombinant protein into a biologically active conformation. The present invention also provides a novel mixed disulfide intermediate wherein the free sulfhydryl groups of reduced recombinant platelet-derived growth factor are derivatized to a disulfide blocking agent.