SARS-CoV-2 Spike Protein ELISA Protocol

A standard direct ELISA protocol was applied essentially as described [18 (link)]. Microtiter plates were coated using 0.1 μg/well of recombinant SARS-CoV-2 spike, S1 domain, RBD, NTD subunits or huACE2 (Sino Biological, Beijing, China). AP-conjugated anti-rabbit IgG or AP-conjugated anti-human IgG (Jackson ImmunoResearch Laboratories, West Grove, PA, USA) were applied for rabbit sera or recombinant antibodies, respectively. Detection was performed using PNPP substrate (Sigma, Rehovot, Israel). Data points were fitted using non-linear regression (Prism 5, GraphPad, San Diego, CA, USA).

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Makdasi E., Levy Y., Alcalay R., Noy-Porat T., Zahavy E., Mechaly A., Epstein E., Peretz E., Cohen H., Bar-On L., Chitlaru T., Cohen O., Glinert I., Achdout H., Israely T., Rosenfeld R, & Mazor O. (2021). Neutralizing Monoclonal Anti-SARS-CoV-2 Antibodies Isolated from Immunized Rabbits Define Novel Vulnerable Spike-Protein Epitope. Viruses, 13(4), 566.

Publication 2021

AP-conjugated anti-rabbit IgG PNPP substrate Prism 5

Anti igg Antibodies Biological Elisa Human Pnpp Prism Rabbit Sars cov 2 Sera Sino Subunits

Corresponding Organization : Israel Institute for Biological Research

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Variable analysis

independent variables

Concentration of recombinant SARS-CoV-2 spike, S1 domain, RBD, NTD subunits or huACE2 used to coat the microtiter plates

dependent variables

Binding of rabbit sera or recombinant antibodies to the coated antigens, as detected by AP-conjugated anti-rabbit IgG or AP-conjugated anti-human IgG and PNPP substrate

control variables

Standard direct ELISA protocol as described in reference [18]
Microtiter plate coating method
Antibody and substrate used for detection

positive controls

None specified

negative controls

None specified

Annotations

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