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Synthesis and Assembly of Hepatitis B Virus-Like Particles in a Pichia pastoris Cell-Free System.

Spice, AJ; Aw, R; Bracewell, DG; Polizzi, KM; (2020) Synthesis and Assembly of Hepatitis B Virus-Like Particles in a Pichia pastoris Cell-Free System. Frontiers in Bioengineering and Biotechnology , 8 , Article 72. 10.3389/fbioe.2020.00072. Green open access

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Abstract

Virus-like particles (VLPs) are supramolecular protein assemblies with the potential for unique and exciting applications in synthetic biology and medicine. Despite the attention VLPs have gained thus far, considerable limitations still persist in their production. Poorly scalable manufacturing technologies and inconsistent product architectures continue to restrict the full potential of VLPs. Cell-free protein synthesis (CFPS) offers an alternative approach to VLP production and has already proven to be successful, albeit using extracts from a limited number of organisms. Using a recently developed Pichia pastoris-based CFPS system, we have demonstrated the production of the model Hepatitis B core antigen VLP as a proof-of-concept. The VLPs produced in the CFPS system were found to have comparable characteristics to those previously produced in vivo and in vitro. Additionally, we have developed a facile and rapid synthesis, assembly and purification methodology that could be applied as a rapid prototyping platform for vaccine development or synthetic biology applications. Overall the CFPS methodology allows far greater throughput, which will expedite the screening of optimal assembly conditions for more robust and stable VLPs. This approach could therefore support the characterization of larger sample sets to improve vaccine development efficiency.

Type: Article
Title: Synthesis and Assembly of Hepatitis B Virus-Like Particles in a Pichia pastoris Cell-Free System.
Location: Switzerland
Open access status: An open access version is available from UCL Discovery
DOI: 10.3389/fbioe.2020.00072
Publisher version: https://doi.org/10.3389/fbioe.2020.00072
Language: English
Additional information: © 2020 Spice, Aw, Bracewell and Polizzi. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
Keywords: Pichia pastoris, cell-free protein synthesis, hepatitis B core antigen, synthetic biology, virus-like particles
UCL classification: UCL
UCL > Provost and Vice Provost Offices > UCL BEAMS
UCL > Provost and Vice Provost Offices > UCL BEAMS > Faculty of Engineering Science
UCL > Provost and Vice Provost Offices > UCL BEAMS > Faculty of Engineering Science > Dept of Biochemical Engineering
URI: https://discovery.ucl.ac.uk/id/eprint/10093696
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