ELISA assays were used to investigate the antibodies´ ability to inhibit the binding of RBD and full S protein to hACE2. For this purpose, 96‐well ELISA plates were coated at 4 °C with 158 nm RBD or 37 nm full S protein, washed and blocked with PBS containing 10% fetal calf serum (FCS). Antibodies were then added at different dilutions (starting from 340 nm and serially diluted 1:3) and incubated for 1 h at 25°C; after washing, hACE2–mouse Fc was added at a constant saturating concentration (160 nm ) and left for 1 h at 25°C. After further washing, bound hACE2 was detected using standard protocols with goat anti‐mouse IgG coupled to alkaline phosphatase (dilution 1:500, SouthernBiotech). After addition of p‐Nitrophenyl Phosphate (PNPP) substrate, colorimetric changes in the ELISA plates were measured at 405 nm with the reader software Gen5 version 1.11.5 (BioTek Instruments). The data were analyzed with GraphPad Prism version 8.4.2.
Gen5 version 1
Manufactured by Agilent Technologies
Gen5 version 1.11.5 is a software package designed for data analysis and instrument control. It provides a user interface for configuring and operating Agilent Technologies' laboratory equipment.
Automatically generated - may contain errors
Lab products found in correlation
2 protocols using gen5 version 1
1
ELISA-based Antibody Inhibition Assay
2
Quantifying SARS-CoV-2 Antibody Binding Inhibition
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ELISAs were used to investigate the antibody’s ability to inhibit the binding of RBD and full S protein to hACE2. We used 96-well ELISA plates coated at 4 °C with 158 nM RBD or 37 nM full S protein, washed and blocked with PBS + 10% fetal calf serum (FCS). Antibodies were then added at different dilutions (starting from 340 nM and serially diluted 1 to 3) and incubated for 1 hour at 25 °C; after washing, hACE2–mouse Fc was added at a constant saturating concentration (160 nM) and left for 1 hour at 25 °C. After further washing, bound hACE2 was detected using standard protocols with goat anti-mouse IgG coupled to alkaline phosphatase (dilution 1:500, Southern Biotech). ELISA plates were measured with the reader software Gen5 version 1.11.5 (BioTek Instruments). Data were analyzed with GraphPad Prism version 8.4.2.
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