Title: A process for recovering a recombinant protein, in biologically active form, from a solution containing inactive protein
Application Number: 93108052 Application Date: 1993.07.01
Publication Number: 1083071 Publication Date: 1994.03.02
Approval Pub. Date: 1998.07.29 Granted Pub. Date: 1998.07.29
International Classifi-cation: C07K1/113;C07K14/61
Applicant(s) Name: Pitman-Moore, Inc. Address:
Inventor(s) Name: Eunkyu Lee;Doreen A. Svihla;Ren-Der Yang
Attorney & Agent: JIANG JIANCHENG WANG YANG
Abstract:
    A process for obtaining a recombinant protein in its natural and bioactive form from a solution containing inactive protein, present in the form of either an insoluble inclusion body or oxidized, misfolded and aggregated waste protein comprises diluting the solution of inactive protein with a first alkaline buffer solution, adding SDS and L-cysteine to simultaneously solubilize and reduce the protein, decreasing the concentration of the SDS and L-cysteine by about 70-90 % by diafiltration against a second alkaline buffer solution, and oxidizing the denatured and reduced protein to form disulfide bonds which correspond to those present in the naturally occurring bioactive protein.
Time: 5