V plex sars cov 2 panel 23 igg kit

Serum samples were analyzed for IgG, IgA, and IgM Abs binding to SARS-CoV-2 proteins. Additionally, sequenced BCR repertoires of the patients were used to design human IgG1 mAbs (produced by Biointron).
Binding to a panel of respiratory viruses was analyzed using a multiplex electrochemiluminescence assay including SARS-CoV-2 S, RBD, NTD, and N antigens, as well as SARS-CoV-1, MERS, OC43, and HKU1 S antigens and Influenza H3 antigen (V-PLEX COVID-19 Coronavirus Panel 1 [IgG, IgM, IgA] Kit, Meso Scale Discovery). SARS-CoV-2–specific Abs were further analyzed for binding to S antigen of several SARS-CoV-2 variants (V-PLEX SARS-CoV-2 Panel 23 [IgG] Kit, Meso Scale Discovery). For read-outs, we used the Meso Quickplex SQ 120 reader (Meso Scale Discovery).
Serum and plasma samples were analyzed at a 1:5,000 and/or 1:50,000 dilution, and mAbs were analyzed at 2–4 different concentrations, ranging from 0.01 μg/mL to 10 μg/mL. For analysis, binding signal was log transformed and normalized, using the normalize function in Prism (GraphPad), with the top binder set to 1.

The V-PLEX SARS-CoV-2 Panel 23 (IgG) kit from Meso Scale Discovery was used to test binding antibodies against the spike protein of the different SARS-CoV-2 VOCs, with serum obtained from hamsters 14 DPI diluted at 10,000X. A standard curve of pooled hamster sera positive for SARS-CoV-2 spike protein was serially diluted 4-fold. To prepare a secondary antibody, a goat anti-hamster IgG cross-adsorbed secondary antibody (ThermoFisher, SA5−10284; each lot is tested to assure specificity and lot-to-lot consistency using an in-house assay. Reference number AB_2868332 (https://www.thermofisher.com/antibody/product/Goat-anti-Syrian-Hamster-IgG-H-L-Cross-Adsorbed-Secondary-Antibody-Polyclonal/SA5-10284) was conjugated using the MSD GOLD SULFO-TAG NHS-Ester Conjugation Pack (MSD). The secondary antibody was diluted 10,000X. The plates were prepped, and samples were run according to the kit’s instruction manual. After the plates were read by the MSD instrument, data was analyzed with the MSD Discovery Workbench Application.

The V-PLEX SARS-CoV-2 Panel 23 (IgG) kit from Meso Scale Discovery was used to test binding antibodies against the spike protein of the different SARS-CoV-2 VOCs, with serum obtained from hamsters 14 DPI diluted at 10,000X. A standard curve of pooled hamster sera positive for SARS-CoV-2 spike protein was serially diluted 4-fold. To prepare a secondary antibody, a goat anti-hamster IgG cross-adsorbed secondary antibody (ThermoFisher) was conjugated using the MSD GOLD SULFO-TAG NHS-Ester Conjugation Pack (MSD). The secondary antibody was diluted 10,000X. The plates were prepped, and samples were run according to the kit’s instruction manual. After the plates were read by the MSD instrument, data was analyzed with the MSD Discovery Workbench Application.