Judy Y. Chang - Hillsborough CA Nancy C. McFarland - Hillsborough CA James R. Swartz - Menlo Park CA
Assignee:
Genentech, Inc. - S. San Francisco CA
International Classification:
C07K 704 C12N 1500
US Classification:
530399
Abstract:
A method for refolding insoluble, improperly folded IGF-I is provided, wherein the IGF-I, precipitated from prokaryotic host cells, it concurrently solubilized, unfolded, and refolded into a biologically active conformation in a single buffer. The buffer contains reducing agent and chaotropic agent to solubilize the IGF-I at concentrations sufficiently low to allow solubilization and folding to occur. Also provided is a triple-protease deficient E. coli host suitable for use in the process.
Method For Refolding Insoluble, Misfolded Insulin-Like Growth Factor-1 Into An Active Conformation
Judy Y. Chang - Hillsborough CA Nancy C. McFarland - Hillsborough CA James R. Swartz - Menlo Park CA
Assignee:
Genentech, Inc. - South San Francisco CA
International Classification:
C12N 1500 C07K 704
US Classification:
530408
Abstract:
A method for refolding insoluble, improperly folded IGF-I is provided, wherein the IGF-I, precipitated from prokaryotic host cells, is concurrently solubilized, unfolded, and refolded into a biologically active conformation in a single buffer. The buffer contains reducing agent and chaotropic agent to solubilize the IGF-I at concentrations sufficiently low to allow solubilization and refolding to occur. Also provided is a triple-protease deficient E. coli host suitable for use in the process.
Nancy C. McFarland - Hillsborough CA James R. Swartz - Menlo Park CA
Assignee:
Genentech, Inc. - South San Francisco CA
International Classification:
C12Q 104 C12Q 102
US Classification:
435 34
Abstract:
An assay is provided for determining the presence of contaminating microbial host cells in a culturing vessel containing a microbial host cell strain that utilizes more than one carbohydrate source as a substrate comprising: (a) culturing the microbial host cell strain in the vessel, wherein the strain comprises nucleic acid encoding a polypeptide of interest and is genetically marked so as not to utilize a carbohydrate source as a substrate; (b) plating an isolated sample of culture from step (a) onto culture media supplemented with a carbohydrate source not utilized by the host cell strain as a substrate; (c) incubating the plated cells at a temperature and for a time sufficient for any positive colony to grow to a detectable level; and (d) detecting whether any colonies are growing on the supplemented culture media.
St. John the Baptist School St. Louis MO 1988-1990, Saint John's Lutheran School Ellisville MO 1990-1994
Community:
Dan Stedman, Jennifer Warfel, Kelly Cary, Amanda Constable, Lauren Weinhold, Christine Eatherton, Steve Manasco, Dave Collora, Jeremy Kenny, Marietta Smith, Jim Nelson, Keith Schmidt