Ieq cov s rbd igg

Mouse and rabbit IgG antibodies against SARS-CoV-2 spike or Ncap were measured by enzyme-linked immunosorbent assays (ELISAs) using precoated ELISA plates (IEQ-CoV-S-RBD-IgG and IEQ-CoV-N-IgG; RayBiotech), according to the manufacturer’s instructions, at RT. Briefly, plasma samples were diluted in sample buffer (RayBiotech), added to antigen-coated wells in triplicates, and incubated at RT for 2 h on a shaker (200 rpm). Commercially available antibody against spike (catalog number S1N-S58; Acro Biosystems) or Ncap (catalog number NUN-S47; Acro Biosystems) was used as a positive control. Plates were washed 3 times with wash buffer and incubated for 1 h at RT with HRP-conjugated goat anti-mouse secondary antibodies (dilution of 1:5,000) (catalog number 115-035-003; Jackson ImmunoResearch) or anti-rabbit antibodies (dilution of 1:5,000) (catalog number 111-035-003; Jackson ImmunoResearch) diluted in assay buffer (RayBiotech). After 3 washes, the plates were developed using the TMB substrate (RayBiotech). After a 15-min incubation, the reaction was stopped by the addition of a stop solution, and the absorbance at 450 nm was recorded using a BioTek Gen5 plate reader (Agilent). Endpoint titers were calculated as the dilution that emitted an optical density (OD) exceeding 4 times that for the PBS control group.

Mouse and rabbit IgG antibodies against SARS-CoV-2 spike or Ncap were measured by enzyme-linked immunosorbent assays (ELISAs) using precoated ELISA plates (IEQ-CoV-S-RBD-IgG and IEQ-CoV-N-IgG; RayBiotech), according to the manufacturer’s instructions, at RT. Briefly, plasma samples were diluted in sample buffer (RayBiotech), added to antigen-coated wells in triplicates, and incubated at RT for 2 h on a shaker (200 rpm). Commercially available antibody against spike (catalog number S1N-S58; Acro Biosystems) or Ncap (catalog number NUN-S47; Acro Biosystems) was used as a positive control. Plates were washed 3 times with wash buffer and incubated for 1 h at RT with HRP-conjugated goat anti-mouse secondary antibodies (dilution of 1:5,000) (catalog number 115-035-003; Jackson ImmunoResearch) or anti-rabbit antibodies (dilution of 1:5,000) (catalog number 111-035-003; Jackson ImmunoResearch) diluted in assay buffer (RayBiotech). After 3 washes, the plates were developed using the TMB substrate (RayBiotech). After a 15-min incubation, the reaction was stopped by the addition of a stop solution, and the absorbance at 450 nm was recorded using a BioTek Gen5 plate reader (Agilent). Endpoint titers were calculated as the dilution that emitted an optical density (OD) exceeding 4 times that for the PBS control group.

Mouse IgG antibody against SARS-CoV-2 antigens was measured by enzyme-linked immunosorbent assays (ELISA) using precoated ELISA plates (IEQ-CoV S RBD-IgG and IEQ-CoVN-IgG, RayBiotech) according to the manufacturer's instructions, at RT. Briefly, mouse plasmas (1:50) were added to preblocked, antigen-coated wells, in duplicate, and incubated at room temperature (RT) for 2 h with gentle rotation (200 rpm). Plates were washed four times with buffer, incubated for 2 h at RT with 1% BSA in PBS, and then with HRP-conjugated goat anti-mouse secondary antibodies (Jackson ImmunoResearch) for 2 h. Wells were washed four times and developed using TMB substrate (RayBiotech). Absorbance at 650 nm was determined for each well using a plate reader (SpectraMaxID3).