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High-yield and scalable cell-free assembly of virus-like particles by dilution

Paper ID Volume ID Publish Year Pages File Format Full-Text
3508 173 2012 9 PDF Available
Title
High-yield and scalable cell-free assembly of virus-like particles by dilution
Abstract

Virus-like particles (VLPs) have been developed as safe and efficacious vaccines, and are increasingly used as carriers for foreign peptide epitopes in modular architectures. An emerging technology for low-cost and rapid-response VLP vaccine manufacture is based on controlled cell-free assembly of capsomeres, which are expressed in Escherichia coli, into VLPs presenting pharmaceutically relevant antigenic modules. A key bioprocessing challenge in this technology is VLP self-assembly, which has until now been studied using qualitative laboratory methods and without sufficient quantitation. In this work, the yield and size distribution of VLPs assembled by dialysis or by ten-fold dilution were compared using quantitative metrics. Membrane-based steps used for dialysis and particle concentration were identified as the key inefficiencies in each method, resulting in 13–18% protein loss. Key inefficiencies were circumvented through process intensification that led to development of a two-fold dilution assembly method. The new process eliminated a unit operation, improved the final concentration of assembly products by a factor of five and reduced buffer consumption nine-fold. Using this process, modular capsomeres presenting a group A Streptococcus (GAS) antigenic module were assembled into high-quality VLPs with a final yield of 54%, which was 18–22 percentage points higher than obtained using conventional methods described in the literature. This study demonstrates the feasibility of manufacturing VLP vaccines in cell-free reactors at high yield, with high structural integrity, using a scalable and simple process. Ongoing development based on these results will be conducive to process-intensified VLP bioprocessing for low-cost vaccine delivery at global scale.

► VLP yield and quality by dialysis and dilution assembly compared quantitatively. ► Process intensification leading to unit operation elimination. ► New process reduces buffer use 9× and increases product concentration 5×. ► VLP yield improved to 54%, 22 percentage points more than conventional methods. ► High-quality VLPs assembled using this scalable, high-yield cell-free method.

Keywords
Virus-like particles; Self-assembly; Bioprocess design; Downstream processing; Process integration; Scale-up
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High-yield and scalable cell-free assembly of virus-like particles by dilution
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Publisher
Database: Elsevier - ScienceDirect
Journal: Biochemical Engineering Journal - Volume 67, 15 August 2012, Pages 88–96
Authors
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Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering
Get Full-Text Now
Don't Miss Today's Special Offer
Price was $35.95
You save - $31
Price after discount Only $4.95
100% Money Back Guarantee
Full-text PDF Download
Online Support
Any Questions? feel free to contact us