A High-throughput Automated ELISA Assay for Detection of IgG Antibodies to the SARS-CoV-2 Spike Protein

Juliana Conkright-Fincham; Chieri Tomomori-Sato; Rich McGhee; Ella M. Leslie; Carolyn J. Beucher; Lauren E. Weems; Shigeo Sato; William B. Redwine; Kyle J. Weaver; Brandon D. Miller; Kym M. Delventhal; John J. Kary; Andrew B. Koebbe; Alexander Dean; Jessica L. Witt; Laura M. Remy; Tari J. Parmely; Chongbei Zhao; Yan Wang; Joan W. Conaway; Jay R. Unruh

doi:10.21769/BioProtoc.4301

A High-throughput Automated ELISA Assay for Detection of IgG Antibodies to the SARS-CoV-2 Spike Protein

Juliana Conkright-Fincham, Chieri Tomomori-Sato, Rich McGhee, Ella M. Leslie, Carolyn J. Beucher, Lauren E. Weems, Shigeo Sato, William B. Redwine, Kyle J. Weaver, Brandon D. Miller, Kym M. Delventhal, John J. Kary, Andrew B. Koebbe, Alexander Dean, Jessica L. Witt, Laura M. Remy, Tari J. Parmely, Chongbei Zhao, Yan Wang, Joan W. ConawayJay R. Unruh

Research output: Contribution to journal › Article › peer-review

Abstract

The SARS-CoV-2 pandemic and vaccination campaign has illustrated the need for high throughput serological assays to quantitatively measure antibody levels. Here, we present a protocol for a high-throughput colorimetric ELISA assay to detect IgG antibodies against the SARS-CoV-2 spike protein. The assay robustly distinguishes positive from negative samples, while controlling for potential non-specific binding from serum samples. To further eliminate background contributions, we demonstrate a computational pipeline for fitting ELISA titration curves, that produces an extremely sensitive antibody signal metric for quantitative comparisons across samples and time.

Original language	English (US)
Article number	e4301
Journal	Bio-protocol
Volume	12
Issue number	2
DOIs	https://doi.org/10.21769/BioProtoc.4301
State	Published - Jan 20 2022
Externally published	Yes

Keywords

COVID-19
ELISA
SARS-CoV-2
Serology
Spike protein

ASJC Scopus subject areas

Biochemistry, Genetics and Molecular Biology(all)
Immunology and Microbiology(all)
Neuroscience(all)
Plant Science

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10.21769/BioProtoc.4301

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Conkright-Fincham, J., Tomomori-Sato, C., McGhee, R., Leslie, E. M., Beucher, C. J., Weems, L. E., Sato, S., Redwine, W. B., Weaver, K. J., Miller, B. D., Delventhal, K. M., Kary, J. J., Koebbe, A. B., Dean, A., Witt, J. L., Remy, L. M., Parmely, T. J., Zhao, C., Wang, Y., ... Unruh, J. R. (2022). A High-throughput Automated ELISA Assay for Detection of IgG Antibodies to the SARS-CoV-2 Spike Protein. Bio-protocol, 12(2), [e4301]. https://doi.org/10.21769/BioProtoc.4301

Conkright-Fincham, J, Tomomori-Sato, C, McGhee, R, Leslie, EM, Beucher, CJ, Weems, LE, Sato, S, Redwine, WB, Weaver, KJ, Miller, BD, Delventhal, KM, Kary, JJ, Koebbe, AB, Dean, A, Witt, JL, Remy, LM, Parmely, TJ, Zhao, C, Wang, Y, Conaway, JW & Unruh, JR 2022, 'A High-throughput Automated ELISA Assay for Detection of IgG Antibodies to the SARS-CoV-2 Spike Protein', Bio-protocol, vol. 12, no. 2, e4301. https://doi.org/10.21769/BioProtoc.4301

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title = "A High-throughput Automated ELISA Assay for Detection of IgG Antibodies to the SARS-CoV-2 Spike Protein",

abstract = "The SARS-CoV-2 pandemic and vaccination campaign has illustrated the need for high throughput serological assays to quantitatively measure antibody levels. Here, we present a protocol for a high-throughput colorimetric ELISA assay to detect IgG antibodies against the SARS-CoV-2 spike protein. The assay robustly distinguishes positive from negative samples, while controlling for potential non-specific binding from serum samples. To further eliminate background contributions, we demonstrate a computational pipeline for fitting ELISA titration curves, that produces an extremely sensitive antibody signal metric for quantitative comparisons across samples and time.",

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author = "Juliana Conkright-Fincham and Chieri Tomomori-Sato and Rich McGhee and Leslie, {Ella M.} and Beucher, {Carolyn J.} and Weems, {Lauren E.} and Shigeo Sato and Redwine, {William B.} and Weaver, {Kyle J.} and Miller, {Brandon D.} and Delventhal, {Kym M.} and Kary, {John J.} and Koebbe, {Andrew B.} and Alexander Dean and Witt, {Jessica L.} and Remy, {Laura M.} and Parmely, {Tari J.} and Chongbei Zhao and Yan Wang and Conaway, {Joan W.} and Unruh, {Jay R.}",

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doi = "10.21769/BioProtoc.4301",

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AU - Conkright-Fincham, Juliana

AU - Tomomori-Sato, Chieri

AU - McGhee, Rich

AU - Leslie, Ella M.

AU - Beucher, Carolyn J.

AU - Weems, Lauren E.

AU - Sato, Shigeo

AU - Redwine, William B.

AU - Weaver, Kyle J.

AU - Miller, Brandon D.

AU - Delventhal, Kym M.

AU - Kary, John J.

AU - Koebbe, Andrew B.

AU - Dean, Alexander

AU - Witt, Jessica L.

AU - Remy, Laura M.

AU - Parmely, Tari J.

AU - Zhao, Chongbei

AU - Wang, Yan

AU - Conaway, Joan W.

AU - Unruh, Jay R.

PY - 2022/1/20

Y1 - 2022/1/20

N2 - The SARS-CoV-2 pandemic and vaccination campaign has illustrated the need for high throughput serological assays to quantitatively measure antibody levels. Here, we present a protocol for a high-throughput colorimetric ELISA assay to detect IgG antibodies against the SARS-CoV-2 spike protein. The assay robustly distinguishes positive from negative samples, while controlling for potential non-specific binding from serum samples. To further eliminate background contributions, we demonstrate a computational pipeline for fitting ELISA titration curves, that produces an extremely sensitive antibody signal metric for quantitative comparisons across samples and time.

AB - The SARS-CoV-2 pandemic and vaccination campaign has illustrated the need for high throughput serological assays to quantitatively measure antibody levels. Here, we present a protocol for a high-throughput colorimetric ELISA assay to detect IgG antibodies against the SARS-CoV-2 spike protein. The assay robustly distinguishes positive from negative samples, while controlling for potential non-specific binding from serum samples. To further eliminate background contributions, we demonstrate a computational pipeline for fitting ELISA titration curves, that produces an extremely sensitive antibody signal metric for quantitative comparisons across samples and time.

KW - COVID-19

KW - ELISA

KW - SARS-CoV-2

KW - Serology

KW - Spike protein

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A High-throughput Automated ELISA Assay for Detection of IgG Antibodies to the SARS-CoV-2 Spike Protein

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Keywords

ASJC Scopus subject areas

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