Reactive diafiltration for assembly and formulation of virus-like particles

Liew, Mervyn W. O., Chuan, Yap P. and Middelberg, Anton P. J. (2012) Reactive diafiltration for assembly and formulation of virus-like particles. Biochemical Engineering Journal, 68 120-128. doi:10.1016/j.bej.2012.07.009

Author Liew, Mervyn W. O.
Chuan, Yap P.
Middelberg, Anton P. J.
Title Reactive diafiltration for assembly and formulation of virus-like particles
Journal name Biochemical Engineering Journal   Check publisher's open access policy
ISSN 1369-703X
Publication date 2012-10
Sub-type Article (original research)
DOI 10.1016/j.bej.2012.07.009
Volume 68
Start page 120
End page 128
Total pages 9
Place of publication Amsterdam, Netherlands
Publisher Elsevier
Collection year 2013
Language eng
Abstract An emerging paradigm to quickly and cost-effectively manufacture virus-like particle (VLP) biopharmaceuticals involves VLP subunit production in highly productive bacterial hosts, followed by assembly in a controlled cell-free environment. Study of cell-free VLP assembly has yet to progress beyond laboratory-based dialysis methods, which are slow and buffer intensive, and are thus difficult to scale. In this study, a scalable cell-free VLP assembly and formulation process using a single diafiltration unit was developed. A reactive constant volume diafiltration VLP assembly process was systematically and quantitatively designed to maximize VLP yield. Using a dilute concentration of capsomeres during VLP assembly minimized membrane-induced aggregation. Optimizing the assembly buffer exchange rate, and the rapid introduction of calcium ions to the assembly mix, facilitated the propitious assembly of VLPs. An absolute increase in VLP yield of 42-56%, compared to initial laboratory-scale batch diafiltration assembly processes, was achieved. The optimized process produced high-quality VLPs (PDI = 1.08 ± 0.01) at high yield (78%). The constant volume diafiltration method has the added advantage of being scalable and represents a high level of process intensification. The methodology reported in this study, based on quantitative analysis of the influences of process changes, is consistent with modern engineering approaches, and can ultimately lead to cheaper and quicker VLP manufacturing routes and hence the accelerated translation of products from laboratory settings.
Keyword Bioprocess design
Downstream processing
Process integration
Downstream processing
Process integration
Virus-like particles
Q-Index Code C1
Q-Index Status Confirmed Code
Institutional Status UQ
Additional Notes Available online 20 July 2012.

Document type: Journal Article
Sub-type: Article (original research)
Collections: Official 2013 Collection
Australian Institute for Bioengineering and Nanotechnology Publications
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