Detergent-mediated liberation of intracellular recombinant virus-like particles (VLPs) from S. cerevisiae homogenate

BIOT 436

Gaik Sui Kee, ucbegsk@ucl.ac.uk1, Hari Pujar, hari_pujar@merck.com2, and Nigel Titchener-Hooker, nigelth@ucl.ac.uk1. (1) Department of Biochemical Engineering, University College London, Torrington Place, London, WC1E 7JE, United Kingdom, (2) Bioprocess R & D, Merck & Co. Inc, P. O. Box 4, Mailstop: WP17-301, West Point, PA 19486
Virus-like Particles (VLPs), such as the hepatitis B surface antigen, are expressed intracellularly in S. cerevisiae. In the recovery of VLPs, the detergent-facilitated release from the host membrane components is a critical process step, responsible for setting the backdrop for subsequent purification. Based on a lipoprotein VLP model, the effect of different detergent conditions were investigated based on tradeoffs between maximal product recovery and preservation of particle antigenicity. Our studies identified that higher concentrations of detergent favours VLP recovery only to a specific threshold. Beyond this, further increase in detergent results in delipidation of VLP, leading to antigenicity loss. Release of contaminating proteins and host lipids into the process stream is also influenced by conditions of the detergent step. The effects of resulting process stream on subsequent unit operations were also characterized.
 

Downstream Processing: Non-Chromatographic Separation Techniques: Improving Process Throughput
8:00 AM-11:00 AM, Thursday, August 23, 2007 BCEC -- 107B, Oral

Division of Biochemical Technology

The 234th ACS National Meeting, Boston, MA, August 19-23, 2007