Accession Number : ADA322790

Title :   Fermentation, Recover, and Purification of the Hc Fragment of the Botulinum Neurotoxin from Pichia pastoris.

Descriptive Note : Annual rept. ,

Corporate Author : NEBRASKA UNIV LINCOLN

Personal Author(s) : Meagher, Michael M.

PDF Url : ADA322790

Report Date : MAR 1997

Pagination or Media Count : 51

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.

Descriptors :   *FERMENTATION, *BACTERIAL TOXINS, *CLOSTRIDIUM BOTULINUM, *NEUROTOXINS, PURIFICATION, PRECIPITATION, METHANOLS, AMMONIUM COMPOUNDS, SULFATES, FEEDING.

Subject Categories : Biochemistry
      Microbiology

Distribution Statement : APPROVED FOR PUBLIC RELEASE