Journal: Med (New York, N.y.)

Article Title: Omicron neutralizing antibody response following booster vaccination compared with breakthrough infection

doi: 10.1016/j.medj.2022.09.001

Figure Lengend Snippet:

Article Snippet: EIA/RIA high binding ELISA plates , Corning , Ref #359096.

Techniques: Virus, Recombinant, Software, Binding Assay, Enzyme-linked Immunosorbent Assay

Journal: Biotechnology Reports

Article Title: Immunogenic fusion proteins induce neutralizing SARS-CoV-2 antibodies in the serum and milk of sheep

doi: 10.1016/j.btre.2023.e00791

Figure Lengend Snippet: Design and characterization of ovine-specific SARS-CoV-2 antigen fusions. a Schematic of designs at the DNA level showing C-terminal immunostimulatory domains IgG2a Fc or GM-CSF. b Schematic of designs at the protein level showing dimerization of the IgG2a Fc fusion proteins. c Summary table showing antigen purity as determined by SDS-PAGE gel followed by densitometry. Purities are listed with standard error calculated from three replicates loaded at 1, 2, and 4 μg of antigen. d ELISA analysis of antigen binding to soluble hACE2 HRP receptors with Ovalbumin used as a negative control.

Article Snippet: High-binding ELISA plates (Corning, #3590) were coated by overnight incubation at 4 °C with 100 μL prepared RBD-His-antigen (1 μg/mL in 0.05 M carbonate bicarbonate buffer, pH 9.6).

Techniques: SDS Page, Enzyme-linked Immunosorbent Assay, Binding Assay, Negative Control

Journal: Biotechnology Reports

Article Title: Immunogenic fusion proteins induce neutralizing SARS-CoV-2 antibodies in the serum and milk of sheep

doi: 10.1016/j.btre.2023.e00791

Figure Lengend Snippet: Evaluation of anti-SARS-CoV-2 antibody responses in ovine serum following immunization with designed antigens. a Schematic showing the design of a 16 week ovine trial to test designed antigen immunogenicity. Hoggets were randomly allocated to groups ( n = 7), immunized once and boosted twice with SARS-CoV-2 antigens over a six week period . Adjuvant-only and Ovalbumin groups were included as negative controls. Blood was periodically sampled for serum analysis until 16 weeks from the first immunization. b Animal weights in each group during the immunization phase of the trial. c ELISA analysis of anti-SARS-CoV-2 RBD (Wuhan) titers for each group during the trial. No detectable response was observed for either Adjuvant or Ovalbumin control groups. Error bars show the standard error within each group. ANOVA testing was used to measure the statistical significance between groups. * p < 0.05, ** p < 0.01, *** p < 0.005 are shown only for the RBD IgG2a Fc group relative to the RBD group.

Article Snippet: High-binding ELISA plates (Corning, #3590) were coated by overnight incubation at 4 °C with 100 μL prepared RBD-His-antigen (1 μg/mL in 0.05 M carbonate bicarbonate buffer, pH 9.6).

Techniques: Immunopeptidomics, Adjuvant, Enzyme-linked Immunosorbent Assay, Control

Journal: Biotechnology Reports

Article Title: Immunogenic fusion proteins induce neutralizing SARS-CoV-2 antibodies in the serum and milk of sheep

doi: 10.1016/j.btre.2023.e00791

Figure Lengend Snippet: Analysis of anti-SARS-CoV-2 antibodies trafficked into colostrum and milk. a Schematic showing the continuation of the ovine trial with select groups. Sheep were mated before a single immunization booster with blood and colostrum/milk samples collected immediately postpartum followed by periodic sampling over 32 days. b ELISA analysis of anti-SARS-CoV-2 RBD (Wuhan) titers in serum for each group over both phases of the ovine trial with black arrows indicating immunization timepoints. c ELISA analysis of anti-SARS-CoV-2 RBD titers in first colostrum and milk. d Neutralizing antibody titers in first colostrum by sVNT at day 0 postpartum. The point in red was not included in the analysis due to the animal being euthanized in the trial. e, f Correlation plots of log transformed anti-SARS-CoV-2 RBD serum titer data against anti-SARS-CoV-2 RBD colostrum/milk data ( e ) and log transformed anti-SARS-CoV-2 RBD colostrum/milk titer data against anti-SARS-CoV-2 colostrum/milk neutralization titer data from the sVNT ( f ). The r value indicates the Pearson correlation coefficient. g Overview of the change in immunoglobulin subclass composition in colostrum/milk between days 0 and 4 postpartum as measured by mass spectrophotometry. Error bars for all plots show the standard error of each group.

Article Snippet: High-binding ELISA plates (Corning, #3590) were coated by overnight incubation at 4 °C with 100 μL prepared RBD-His-antigen (1 μg/mL in 0.05 M carbonate bicarbonate buffer, pH 9.6).

Techniques: Sampling, Enzyme-linked Immunosorbent Assay, Transformation Assay, Neutralization, Spectrophotometry

Journal: Biotechnology Reports

Article Title: Immunogenic fusion proteins induce neutralizing SARS-CoV-2 antibodies in the serum and milk of sheep

doi: 10.1016/j.btre.2023.e00791

Figure Lengend Snippet: Evaluation of SARS-CoV-2 neutralizing responses in serum by surrogate viral neutralization testing. a Cartoon showing the principle of the surrogate viral neutralization test. Neutralization generates a loss of ELISA signal as hACE2-HRP binding to SARS-CoV-2 RBD is reduced by antibody blocking, with such approaches comparable in specificity and sensitivity to live viral assays [ , ]. b Plots showing relative inhibitory concentrations for RBD interaction with hACE2 for each trial group at weeks 6 and 9 as measured by serum dilution factor. LY-CoV555 is the positive control anti-SARS-CoV2 clinical antibody bamlanivimab diluted from 6.9 μg/mL. Error bars show the standard error of each group. ANOVA testing was used to measure the statistical significance between groups with * p < 0.05, ** p < 0.01, and *** p < 0.005. c Correlation plot of log transformed anti-SARS-CoV-2 neutralization titer data from the sVNT vs serum titer data against SARS-CoV-2 RBD for weeks 6 and 9. Shaded areas indicate the 95% confidence intervals for power law fitting.

Article Snippet: High-binding ELISA plates (Corning, #3590) were coated by overnight incubation at 4 °C with 100 μL prepared RBD-His-antigen (1 μg/mL in 0.05 M carbonate bicarbonate buffer, pH 9.6).

Techniques: Neutralization, Enzyme-linked Immunosorbent Assay, Binding Assay, Blocking Assay, Positive Control, Transformation Assay