1. A scalable and high yielding SARS-CoV-2 spike protein receptor binding domain production process.
- Author
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Riguero, Valeria, Delmar, Jared, Dippel, Andrew, McTamney, Patrick, Luo, Ethan, Martinez, Antonio, Ren, Kuishu, van Dyk, Nydia, and O'Connor, Ellen
- Subjects
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SARS-CoV-2 , *PROTEIN receptors , *PROTEIN binding , *CARRIER proteins , *MANUFACTURING processes , *PROTEIN-protein interactions - Abstract
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. [ABSTRACT FROM AUTHOR]
- Published
- 2023
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