Inventors:
David H. Reifsnyder - El Cerrito CA, US
Duane Inlow - Alamo CA, US
Glenn Dorin - San Rafael CA, US
Patricio T. Riquelme - Walnut Creek CA, US
Cynthia A. Cowgill - Berkeley CA, US
Douglas G. Bolesch - Berkeley CA, US
Mark E. Gustafson - St. Charles MO, US
Assignee:
Novartis Vaccines and Diagnostics, Inc. - Emeryville CA
Pharmacia & Upjohn Company - Kalamazoo MI
International Classification:
C07K 14/00
Abstract:
Highly purified preparations of TFPI or TFPI analogs can be prepared using a method that generally involves the following steps: (1) expression of TFPI or TFPI analog in , (2) isolation of refractile bodies, (3) dissolution of the refractile bodies and refolding of the expressed TFPI or TFPI analog, (4) SP-Sepharose fast flow (FF) chromatography, (5) a first concentration and diafiltration step, (6) Q-Sepharose high (HP) performance chromatography, (7) butyl hydrophobic interaction chromatography (HIC), (8) SP-Sepharose HP chromatography, and (9) a second concentration/diafiltration step. Less than about 12% of the TFPI or TFPI analog molecules in such preparations are modified TFPI or TFPI analog species (i. e. , oxidized, carbamylated, acetylated, deamidated, aggregated, or misfolded species).