V plex sars cov 2 panel 1 kit

SARS-CoV-2 wild-type strain S glycoprotein RBD- and NTD-coupled magnetic beads (Acro Biosystems, Cat.no. MBS-K002 and MBS-K019; 40 μg RBD/mg beads and 38 μg NTD/mg beads, respectively) were prepared according to the manufacturer’s instructions. Beads were resuspended in ultrapure water at 1 mg beads/mL and a magnet was used to collect and wash the beads with PBS. Beads were resuspended in serum to obtain 20 μg RBD- or NTD-bait per 100 μL serum. A mock depletion (undepleted control) was performed for each serum by adding 0.5 mg Biotin-saturated MyOne Streptavidin T1 Dynabeads (ThermoFisher, Cat.no. 65601) per 100 μL serum. Beads were incubated with human sera for 1 hour with gentle rotation. A magnet was used to separate bead-bound antibodies from the depleted supernatant. Depleted and undepleted sera were analyzed for cross-neutralization capacity using pseudovirus neutralization assays. Depletion efficacy for both RBD- and NTD-binding antibodies was determined by a multiplexed electrochemiluminescence immunoassay (Meso Scale Discovery, V-Plex SARS-CoV-2 Panel 1 Kit, Cat. No. K15359U-2). Table of the neutralization titers is provided (Table S11).

IgM and IgG titers were quantified from serum collected for all animals on Days 0, 7, 14, 28, 42, and 49. Meso Scale Discovery V-PLEX SARS-CoV-2 Panel 1 Kit (Rockville, MD) was used for determining antibody binding for IgG (K15359U) and IgM (K15360U) specific for SARS-CoV-2 S1 RBD. Protocols were followed as per manufacture’s recommendations. All diluted samples fell within standard curve ranges. A Meso Scale Discovery® MESO QuickPlex SQ 120 instrument was used for measuring chemiluminescence with Methodical Minds ^™ acquisition software. Analysis was performed on the DISCOVERY WORKBENCH® software (version 4.0). Concentrations are relative to internal controls and were reported as arbitrary units/mL (AU/mL). Graphpad Prism 8 was used for preparation of graphs and statistical analysis.