A fast, efficient, and scalable method for purifying recombinant SARS-CoV-2 spike protein.
J Chromatogr B Analyt Technol Biomed Life Sci
; 1215: 123579, 2023 Jan 15.
Article
in English
| MEDLINE | ID: covidwho-2239446
ABSTRACT
Recombinant SARS-CoV-2 trimeric spike protein produced by mammalian cell culture is a potential candidate for a COVID-19 vaccine. However, this protein is much larger than most typical biopharmaceutical proteins and its large-scale manufacture is therefore challenging. Particularly, its purification using resin-based chromatography is difficult as the diffusive transport of this protein to and from its binding site within the pores of the stationary phase particles is slow. Therefore, very low flow rates need to be used during binding and elution, and this slows down the purification process. Also, due to its large size, the binding capacity of this protein on resin-based media is low. Membrane chromatography is an efficient and scalable technique for purifying biopharmaceuticals. The predominant mode of solute transport in a membrane is convective and hence it is considered better than resin-based chromatography for purifying large proteins. In this paper, we propose a membrane chromatography-based purification method for fast and scalable manufacture of recombinant SARS-CoV-2 trimeric spike protein. A combination of cation exchange z2 laterally-fed membrane chromatography and size exclusion chromatography was found to be suitable for obtaining a homogeneous spike protein sample from mammalian cell culture supernatant. The proposed method is both fast and scalable and could be explored as a method for manufacturing vaccine grade spike protein.
Keywords
Full text:
Available
Collection:
International databases
Database:
MEDLINE
Main subject:
Recombinant Proteins
/
Spike Glycoprotein, Coronavirus
Topics:
Vaccines
Language:
English
Journal:
J Chromatogr B Analyt Technol Biomed Life Sci
Journal subject:
Biomedical Engineering
Year:
2023
Document Type:
Article
Affiliation country:
J.jchromb.2022.123579
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