Improved production of SARS-CoV-2 spike receptor-binding domain (RBD) for serology assays

Author:

Mehalko,Jennifer

Drew, Matthew

Snead,Kelly

Denson,John-Paul

Wall,Vanessa

Taylor,Troy

Sadtler, Kaitlyn

Messing,Simon

Gillette, William

Esposito,Dom
Author Address

Protein Expression Laboratory, NCI RAS Initiative, Frederick National Laboratory for Cancer Research, Leidos Biomedical Research, Inc., Frederick, MD 21702., Section on Immuno-Engineering, National Institute for Biomedical Imaging and Bioengineering, National Institutes of Health, Bethesda, MD 20894., Protein Expression Laboratory, NCI RAS Initiative, Frederick National Laboratory for Cancer Research, Leidos Biomedical Research, Inc., Frederick, MD 21702. Electronic address: dom.esposito@nih.gov.,

Abstract:

The receptor-binding domain (RBD) of the SARS-CoV-2 spike protein is a commonly used antigen for serology assays critical to determining the extent of SARS-CoV-2 exposure in the population. Different versions of the RBD protein have been developed and utilized in assays, with higher sensitivity attributed to particular forms of the protein. To improve the yield of these high-sensitivity forms of RBD and support the increased demand for this antigen in serology assays, we investigated several protein expression variables including DNA elements such as promoters and signal peptides, cell culture expression parameters, and purification processes. Through this investigation, we developed a simplified and robust purification strategy that consistently resulted in high levels of the high-sensitivity form of RBD and demonstrated that a carboxyterminal tag is responsible for the increased sensitivity in the ELISA. These improved reagents and processes produce high-quality proteins which are functional in serology assays and can be used to investigate seropositivity to SARS-CoV-2 infection. Copyright © 2020. Published by Elsevier Inc.

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