High-throughput process development of an alternative platform for the production of virus-like particles in Escherichia coli

Ladd Effio, Christopher, Baumann, Pascal, Weigel, Claudia, Vormittag, Philipp, Middelberg, Anton and Hubbuch, Jürgen (2016) High-throughput process development of an alternative platform for the production of virus-like particles in Escherichia coli. Journal of Biotechnology, 219 7-19. doi:10.1016/j.jbiotec.2015.12.018

Attached Files (Some files may be inaccessible until you login with your UQ eSpace credentials)
Name Description MIMEType Size Downloads
UQ376717_OA.pdf Full text (open access) application/pdf 3.93MB 0

Author Ladd Effio, Christopher
Baumann, Pascal
Weigel, Claudia
Vormittag, Philipp
Middelberg, Anton
Hubbuch, Jürgen
Title High-throughput process development of an alternative platform for the production of virus-like particles in Escherichia coli
Formatted title
High-throughput process development of an alternative platform for the production of virus-like particles in Escherichia coli
Journal name Journal of Biotechnology   Check publisher's open access policy
ISSN 1873-4863
0168-1656
Publication date 2016-02-10
Year available 2015
Sub-type Article (original research)
DOI 10.1016/j.jbiotec.2015.12.018
Open Access Status File (Author Post-print)
Volume 219
Start page 7
End page 19
Total pages 13
Place of publication Amsterdam, Netherlands
Publisher Elsevier
Language eng
Formatted abstract
The production of safe vaccines against untreatable or new diseases has pushed the research in the field of virus-like particles (VLPs). Currently, a large number of commercial VLP-based human vaccines and vaccine candidates are available or under development. A promising VLP production route is the controlled in vitro assembly of virus proteins into capsids.

In the study reported here, a high-throughput screening (HTS) procedure was implemented for the upstream process development of a VLP platform in bacterial cell systems. Miniaturized cultivations were carried out in 48-well format in the BioLector system (m2p-Labs, Germany) using an Escherichia coli strain with a tac promoter producing the murine polyomavirus capsid protein (VP1). The screening procedure incorporated micro-scale cultivations, HTS cell disruption by sonication and HTS-compatible analytics by capillary gel electrophoresis. Cultivation temperatures, shaking speeds, induction and medium conditions were varied to optimize the product expression in E. coli. The most efficient system was selected based on an evaluation of soluble and insoluble product concentrations as well as on the percentage of product in the total soluble protein fraction. The optimized system was scaled up to cultivation 2.5 L shaker flask scale and purified using an anion exchange chromatography membrane adsorber, followed by a size exclusion chromatography polishing procedure. For proof of concept, purified VP1 capsomeres were assembled under defined buffer conditions into empty capsids and characterized using transmission electron microscopy (TEM).

The presented HTS procedure allowed for a fast development of an efficient production process of VLPs in E. coli. Under optimized cultivation conditions, the VP1 product totalled up to 43% of the total soluble protein fraction, yielding 1.63 mg VP1 per mL of applied cultivation medium. The developed production process strongly promotes the murine polyoma-VLP platform, moving towards an industrially feasible technology for new chimeric vaccines.
Keyword BioLector
Escherichia coli
High-throughput screening
Micro-scale cultivation
Virus-like particles
Q-Index Code C1
Q-Index Status Provisional Code
Institutional Status UQ
Additional Notes Published online 17 December 2015

Document type: Journal Article
Sub-type: Article (original research)
Collections: Official 2016 Collection
Australian Institute for Bioengineering and Nanotechnology Publications
 
Versions
Version Filter Type
Citation counts: TR Web of Science Citation Count  Cited 0 times in Thomson Reuters Web of Science Article
Scopus Citation Count Cited 3 times in Scopus Article | Citations
Google Scholar Search Google Scholar
Created: Tue, 12 Jan 2016, 10:18:43 EST by System User on behalf of Learning and Research Services (UQ Library)