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Optimizing high-yield production of SARS-CoV-2 soluble spike trimers for serology assays

  1. Author:
    Esposito,Dom
    Mehalko,Jennifer
    Drew, Matthew
    Snead,Kelly
    Wall,Vanessa
    Taylor,Troy
    Frank,Peter
    Denson,John-Paul
    Hong,Min
    Gulten,Gulcin
    Sadtler, Kaitlyn
    Messing,Simon
    Gillette,Bill

  2. Author Address

    Protein Expression Laboratory, NCI RAS Initiative, Cancer Research Technology Program, Frederick National Laboratory for Cancer Research, Leidos Biomedical Research, Inc., Frederick, MD, 21702, USA. Electronic address: dom.esposito@nih.gov., Section on Immuno-Engineering, National Institute for Biomedical Imaging and Bioengineering, National Institutes of Health, Bethesda, MD, 20894, USA.,
    1. Year: 2020
    2. Date: OCT
    3. Epub Date: 2020 06 03
  2. Journal: Protein expression and purification
    1. 174
    2. Pages: 105686
  4. Type of Article: Article
  5. Article Number: 105686
  6. ISSN: 1046-5928
  1. Abstract:

    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. Copyright © 2020. Published by Elsevier Inc.

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External Sources

  1. View Full Text
  2. DOI: 10.1016/j.pep.2020.105686
  3. PMID: 32504802
  4. PMCID: PMC7271859
  5. View record in Web of Science®
  6. WOS: 000571503500011
  7. PII : S1046-5928(20)30273-4

Library Notes

  1. Fiscal Year: FY2019-2020
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