Fermentation, Recover, and Purification of the Hc Fragment of the Botulinum Neurotoxin from Pichia pastoris.
Abstract
This mid-term report is an update on the purification of Clostridium botilinum serotype B Hc fragment expressed intracellularly in Pichia pastoris, clone BBC-C-02136 A5 BoNT B (Hc)/pHILD4. During the first half of this year the fermentation was improved by the addition of casamino acids during the methanol feed. In addition, a process using a Streamline column (fluidized ion exchange column) was used to capture the Hc fragment directly from the cell homogenate at approximately 70 to 80% pure. This step replaces the polyethyelenimine precipitation and the ammonium sulfate precipitation step in the previous purification process. A purification process was completed that generates a 99+% pure product by SDS-PAOE and 97% to 99% pure on isoelectric focusing.
Document Details
- Document Type
- Technical Report
- Publication Date
- Mar 01, 1997
- Accession Number
- ADA322790
Entities
People
- Michael M. Meagher
Organizations
- University of Nebraska–Lincoln