Bio-Protocol (Jan 2022)

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 Leslie,
  • Carolyn Beucher,
  • Lauren Weems,
  • Shigeo Sato,
  • William Redwine,
  • Kyle Weaver,
  • Brandon Miller,
  • Kym Delventhal,
  • John Kary,
  • Andrew Koebbe,
  • Alexander Dean,
  • Jessica Witt,
  • Laura Remy,
  • Tari Parmely,
  • Chongbei Zhao,
  • Yan Wang,
  • Joan Conaway,
  • Jay Unruh

DOI
https://doi.org/10.21769/BioProtoc.4301
Journal volume & issue
Vol. 12, no. 2

Abstract

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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.