Scalable, methanol-free manufacturing of the SARS-CoV-2 receptor-binding domain in engineered Komagataella phaffii.

Autor: Dalvie NC; Department of Chemical Engineering, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA.; Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA., Biedermann AM; Department of Chemical Engineering, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA.; Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA., Rodriguez-Aponte SA; Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA.; Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA., Naranjo CA; Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA., Rao HD; Serum Institute of India Pvt. Ltd., Pune, India., Rajurkar MP; Serum Institute of India Pvt. Ltd., Pune, India., Lothe RR; Serum Institute of India Pvt. Ltd., Pune, India., Shaligram US; Serum Institute of India Pvt. Ltd., Pune, India., Johnston RS; Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA., Crowell LE; Department of Chemical Engineering, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA.; Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA., Castelino S; Department of Chemical Engineering, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA., Tracey MK; Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA., Whittaker CA; Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA., Love JC; Department of Chemical Engineering, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA.; Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA.
Jazyk: angličtina
Zdroj: Biotechnology and bioengineering [Biotechnol Bioeng] 2022 Feb; Vol. 119 (2), pp. 657-662. Date of Electronic Publication: 2021 Nov 15.
DOI: 10.1002/bit.27979
Abstrakt: Prevention of COVID-19 on a global scale will require the continued development of high-volume, low-cost platforms for the manufacturing of vaccines to supply ongoing demand. Vaccine candidates based on recombinant protein subunits remain important because they can be manufactured at low costs in existing large-scale production facilities that use microbial hosts like Komagataella phaffii (Pichia pastoris). Here, we report an improved and scalable manufacturing approach for the SARS-CoV-2 spike protein receptor-binding domain (RBD); this protein is a key antigen for several reported vaccine candidates. We genetically engineered a manufacturing strain of K. phaffii to obviate the requirement for methanol induction of the recombinant gene. Methanol-free production improved the secreted titer of the RBD protein by >5X by alleviating protein folding stress. Removal of methanol from the production process enabled to scale up to a 1200 L pre-existing production facility. This engineered strain is now used to produce an RBD-based vaccine antigen that is currently in clinical trials and could be used to produce other variants of RBD as needed for future vaccines.
(© 2021 The Authors. Biotechnology and Bioengineering published by Wiley Periodicals LLC.)
Databáze: MEDLINE
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