An Endotoxin Binding Protein for Treatment of Septic Shock
Abstract
Native ENP has been extracted from the 1990 blood collection season. The extract has been purified over the first purification column (ion change) and 10% has been processed through the final purification. Recombinant Endotoxin Neutralizing Protein: Fermentation has been scaled from 7 liters to 30 liters. The major improvement in this period has been in the down stream processing. Our extraction/purification procedure has required incubation of the yeast fermentation broth with 3 molar urea to dissociate complexes formed between ENP and uncharacterized medium or cellular components. It was found that changing the medium pH from 5.5 to 6.5 results in both increased secretion of ENP into the broth and elimination of the urea pre-incubation. The omission of urea, however, has created a new need to process the broth faster. Presumably, urea was acting to dissociate complex as well as denature proteases.
Document Details
- Document Type
- Technical Report
- Publication Date
- Feb 15, 1991
- Accession Number
- ADA233382
Entities
People
- Norman R. Wainwright