Improved fusion tag cleavage strategies in the downstream processing of self-assembling virus-like particle vaccines

Connors, Natalie K., Wu, Yang, Lua, Linda H. L. and Middelberg, Anton P. J. (2013) Improved fusion tag cleavage strategies in the downstream processing of self-assembling virus-like particle vaccines. Food and Bioproducts Processing, 92 2: 143-151. doi:10.1016/j.fbp.2013.08.012


Author Connors, Natalie K.
Wu, Yang
Lua, Linda H. L.
Middelberg, Anton P. J.
Title Improved fusion tag cleavage strategies in the downstream processing of self-assembling virus-like particle vaccines
Journal name Food and Bioproducts Processing   Check publisher's open access policy
ISSN 0960-3085
1744-3571
Publication date 2013-09-04
Year available 2013
Sub-type Article (original research)
DOI 10.1016/j.fbp.2013.08.012
Volume 92
Issue 2
Start page 143
End page 151
Total pages 9
Place of publication London, United Kingdom
Publisher Elsevier
Language eng
Abstract Virus-like particle (VLP) vaccines are emerging as an exciting platform for the delivery of antigenic modules to enable engineering of desirable immunological outcomes, both in therapeutic and prophylactic settings. The processing of these macromolecular assemblies raises new challenges not previously encountered for simpler protein therapeutics. Microbial expression of viral precursor protein complexes (capsomeres) and their subsequent cell-free self-assembly represent a new and technologically interesting pathway to VLP vaccines, yet significant scientific and engineering challenges remain. Among these, the use of thrombin within the existing laboratory process must be eliminated, for regulatory, cost and product quality reasons. Here, the use of alternatives to thrombin is explored. It is shown that tobacco etch virus protease (TEVp) is a viable alternative to thrombin, and leads to higher-quality VLP product; as TEVp has no known human physiological or biochemical role, its bioprocessing acceptability is expected to be higher than for thrombin. The results presented here enhance the scale-up potential of microbial VLP processing, and suggest that TEVp may be a preferable enzyme for use in other bioprocess settings, including those related to the processing of less complex biologics.
Keyword Virus-like particles
Thrombin
TEVp
Enzyme
Murine polyomavirus
GST
Q-Index Code C1
Q-Index Status Confirmed Code
Institutional Status UQ
Additional Notes Published online ahead of print 4 September 2013.

Document type: Journal Article
Sub-type: Article (original research)
Collections: Official 2014 Collection
Australian Institute for Bioengineering and Nanotechnology Publications
 
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Created: Wed, 08 Jan 2014, 21:10:22 EST by Linda Lua on behalf of Aust Institute for Bioengineering & Nanotechnology