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Esposito, Dominic; Mehalko, Jennifer; Drew, Matthew; Snead, Kelly; Wall, Vanessa; Taylor, Troy; Frank, Peter; Denson, John-Paul; Hong, Min; Gulten, Gulcin; Sadtler, Kaitlyn; Messing, Simon; Gillette, William
Protein expression and purification, 10/2020, Letnik: 174Journal Article
The SARS-CoV-2 spike trimer is the primary antigen for several serology assays critical to determining the extent of SARS-CoV-2 exposure in the population. Until stable cell lines are developed to increase the titer of this secreted protein in mammalian cell culture, the low yield of spike protein produced from transient transfection of HEK293 cells will be a limiting factor for these assays. To improve the yield of spike protein and support the high demand for antigens in serology assays, we investigated several recombinant protein expression variables by altering the incubation temperature, harvest time, chromatography strategy, and final protein manipulation. Through this investigation, we developed a simplified and robust purification strategy that consistently yields 5 mg of protein per liter of expression culture for two commonly used forms of the SARS-CoV-2 spike protein. We show that these proteins form well-behaved stable trimers and are consistently functional in serology assays across multiple protein production lots. •Improved yields of SARS-CoV-2 spike protein through modification of expression and purification parameters.•Yields of greater than 5 mg/l obtained for VRC spike under optimal conditions.•Spike protein quality was validated by QC methods to ensure utility in serology assays.
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