Quantitative Measurement of Anti-SARS-CoV-2 Antibodies
Plasma samples were diluted 4000-fold, and the total IgG and IgM levels against the SARS-CoV-2 spike protein were measured using a custom Single Molecule Array (Simoa) assay as described [29 (link)] on an automated HD-X Analyzer (Quanterix, Billerica, MA, USA), to provide a quantitative reference for anti-spike antibody titers in the plasma samples. Normalized mean Average Enzymes per Bead (AEB) levels were calculated using a standard set of calibrators produced by serially diluting a large volume of plasma from seroconverted individuals. Antibody concentrations were estimated using a calibration curve of recombinant anti-SARS-CoV-2 antibodies [30 (link)].
Trombetta B.A., Kandigian S.E., Kitchen R.R., Grauwet K., Webb P.K., Miller G.A., Jennings C.G., Jain S., Miller S., Kuo Y., Sweeney T., Gilboa T., Norman M., Simmons D.P., Ramirez C.E., Bedard M., Fink C., Ko J., De León Peralta E.J., Watts G., Gomez-Rivas E., Davis V., Barilla R.M., Wang J., Cunin P., Bates S., Morrison-Smith C., Nicholson B., Wong E., El-Mufti L., Kann M., Bolling A., Fortin B., Ventresca H., Zhou W., Pardo S., Kwock M., Hazra A., Cheng L., Ahmad Q.R., Toombs J.A., Larson R., Pleskow H., Luo N.M., Samaha C., Pandya U.M., De Silva P., Zhou S., Ganhadeiro Z., Yohannes S., Gay R., Slavik J., Mukerji S.S., Jarolim P., Walt D.R., Carlyle B.C., Ritterhouse L.L, & Suliman S. (2021). Evaluation of serological lateral flow assays for severe acute respiratory syndrome coronavirus-2. BMC Infectious Diseases, 21, 580.
Anti antibodies Anti antibody Antibody Assay Enzymes levelsPlasma Plasma volume Sars cov 2 Sars cov 2 spike protein
Corresponding Organization :
Other organizations :
Massachusetts General Hospital, Harvard University, Mass General Brigham, Dana-Farber Cancer Institute, Brigham and Women's Hospital, Center for Cancer Research, Reproductive Science Center, Northeastern University
Total IgG levels against the SARS-CoV-2 spike protein
Total IgM levels against the SARS-CoV-2 spike protein
control variables
Use of a custom Single Molecule Array (Simoa) assay to measure antibody levels
Use of an automated HD-X Analyzer (Quanterix) to perform the measurements
Use of a standard set of calibrators produced by serially diluting a large volume of plasma from seroconverted individuals to calculate normalized mean Average Enzymes per Bead (AEB) levels
Use of a calibration curve of recombinant anti-SARS-CoV-2 antibodies to estimate antibody concentrations
positive controls
Plasma from seroconverted individuals
negative controls
Not explicitly mentioned
Annotations
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