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A scalable and high yielding SARS-CoV-2 spike protein receptor binding domain production process

The Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) spike protein is of interest for the development of vaccines and therapeutics against COVID-19. Vaccines are designed to raise an immune response against the spike protein. Other therapies attempt to block the interaction of the spike...

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Autores principales: Riguero, Valeria, Delmar, Jared, Dippel, Andrew, McTamney, Patrick, Luo, Ethan, Martinez, Antonio, Ren, Kuishu, van Dyk, Nydia, O'Connor, Ellen
Formato: Online Artículo Texto
Lenguaje:English
Publicado: The Authors. Published by Elsevier Inc. 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9890279/
https://www.ncbi.nlm.nih.gov/pubmed/36736512
http://dx.doi.org/10.1016/j.pep.2023.106241
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author Riguero, Valeria
Delmar, Jared
Dippel, Andrew
McTamney, Patrick
Luo, Ethan
Martinez, Antonio
Ren, Kuishu
van Dyk, Nydia
O'Connor, Ellen
author_facet Riguero, Valeria
Delmar, Jared
Dippel, Andrew
McTamney, Patrick
Luo, Ethan
Martinez, Antonio
Ren, Kuishu
van Dyk, Nydia
O'Connor, Ellen
author_sort Riguero, Valeria
collection PubMed
description The Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) spike protein is of interest for the development of vaccines and therapeutics against COVID-19. Vaccines are designed to raise an immune response against the spike protein. Other therapies attempt to block the interaction of the spike protein and mammalian cells. Therefore, the spike protein itself and specific interacting regions of the spike protein are reagents required by industry to enable the advancement of medicines to combat SARS-CoV-2. Early production methods of the SARS-CoV-2 spike protein receptor binding domain (RBD) were labor intensive with scalability challenges. In this work, we describe a high yielding and scalable production process for the SARS-CoV-2 RBD. Expression was performed in human embryonic kidney (HEK) 293 cells followed by a two-column purification process including immobilized metal affinity chromatography (IMAC) followed by Ceramic Hydroxyapatite (CHT). The improved process showed good scalability, enabling efficient purification of 2.5 g of product from a 200 L scale bioreactor.
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spelling pubmed-98902792023-02-01 A scalable and high yielding SARS-CoV-2 spike protein receptor binding domain production process Riguero, Valeria Delmar, Jared Dippel, Andrew McTamney, Patrick Luo, Ethan Martinez, Antonio Ren, Kuishu van Dyk, Nydia O'Connor, Ellen Protein Expr Purif Article The Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) spike protein is of interest for the development of vaccines and therapeutics against COVID-19. Vaccines are designed to raise an immune response against the spike protein. Other therapies attempt to block the interaction of the spike protein and mammalian cells. Therefore, the spike protein itself and specific interacting regions of the spike protein are reagents required by industry to enable the advancement of medicines to combat SARS-CoV-2. Early production methods of the SARS-CoV-2 spike protein receptor binding domain (RBD) were labor intensive with scalability challenges. In this work, we describe a high yielding and scalable production process for the SARS-CoV-2 RBD. Expression was performed in human embryonic kidney (HEK) 293 cells followed by a two-column purification process including immobilized metal affinity chromatography (IMAC) followed by Ceramic Hydroxyapatite (CHT). The improved process showed good scalability, enabling efficient purification of 2.5 g of product from a 200 L scale bioreactor. The Authors. Published by Elsevier Inc. 2023-05 2023-02-01 /pmc/articles/PMC9890279/ /pubmed/36736512 http://dx.doi.org/10.1016/j.pep.2023.106241 Text en © 2023 The Authors Since January 2020 Elsevier has created a COVID-19 resource centre with free information in English and Mandarin on the novel coronavirus COVID-19. The COVID-19 resource centre is hosted on Elsevier Connect, the company's public news and information website. Elsevier hereby grants permission to make all its COVID-19-related research that is available on the COVID-19 resource centre - including this research content - immediately available in PubMed Central and other publicly funded repositories, such as the WHO COVID database with rights for unrestricted research re-use and analyses in any form or by any means with acknowledgement of the original source. These permissions are granted for free by Elsevier for as long as the COVID-19 resource centre remains active.
spellingShingle Article
Riguero, Valeria
Delmar, Jared
Dippel, Andrew
McTamney, Patrick
Luo, Ethan
Martinez, Antonio
Ren, Kuishu
van Dyk, Nydia
O'Connor, Ellen
A scalable and high yielding SARS-CoV-2 spike protein receptor binding domain production process
title A scalable and high yielding SARS-CoV-2 spike protein receptor binding domain production process
title_full A scalable and high yielding SARS-CoV-2 spike protein receptor binding domain production process
title_fullStr A scalable and high yielding SARS-CoV-2 spike protein receptor binding domain production process
title_full_unstemmed A scalable and high yielding SARS-CoV-2 spike protein receptor binding domain production process
title_short A scalable and high yielding SARS-CoV-2 spike protein receptor binding domain production process
title_sort scalable and high yielding sars-cov-2 spike protein receptor binding domain production process
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9890279/
https://www.ncbi.nlm.nih.gov/pubmed/36736512
http://dx.doi.org/10.1016/j.pep.2023.106241
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