Journal: Journal of Immunological Methods

Article Title: Parameters of immunoglobulin extraction from dried blood spot cards and immunoassays for detection of antibody response to pathogens including the novel SARS-CoV-2

doi: 10.1016/j.jim.2021.112996

Figure Lengend Snippet: Immune response to of SARS-CoV-2 recombinant S protein RBD antigen in mice. Titration in antigen-mediated ELISA of 4-fold diluted antibodies extracted from DBS at different study time points (A). Extended titration of the DBS samples collected on day 14 and day 21 compared to serum samples collected on day 21 as measured by antigen-mediated ELISA against spike RBD recombinant protein (B). Extracted samples were run in duplicates and results are presented as mean ± SD (error bars) absorbance for the corresponding dilutions.

Article Snippet: Purified recombinant protein corresponding to the receptor-binding domain (RBD) of SARS-CoV-2 spike protein and derived from human cells (ProSci, Fort Collins CO) was used as antigen for immunization of mice.

Techniques: Recombinant, Titration, Enzyme-linked Immunosorbent Assay

Journal: Journal of Immunological Methods

Article Title: Parameters of immunoglobulin extraction from dried blood spot cards and immunoassays for detection of antibody response to pathogens including the novel SARS-CoV-2

doi: 10.1016/j.jim.2021.112996

Figure Lengend Snippet: Total extracted IgG (in μg/ml) from DBS of the S protein RBD-immunized Ifnarko-CD46Ge mice as measured by ELISA. Range: 54.32–347.84; Mean: 188.92 ± 86.95 per dried spot.

Article Snippet: Purified recombinant protein corresponding to the receptor-binding domain (RBD) of SARS-CoV-2 spike protein and derived from human cells (ProSci, Fort Collins CO) was used as antigen for immunization of mice.

Techniques: Enzyme-linked Immunosorbent Assay

Journal: Journal of Immunological Methods

Article Title: Parameters of immunoglobulin extraction from dried blood spot cards and immunoassays for detection of antibody response to pathogens including the novel SARS-CoV-2

doi: 10.1016/j.jim.2021.112996

Figure Lengend Snippet: Immune response to recombinant S protein RBD antigen in SARS-CoV-2-convalescent individuals. Samples were run in 4-fold dilutions from 1:2 to 1:128 in antigen-mediated ELISA with plates coated with 100 ng per well antigen. The absorbance (OD 450nm ) reading for individual samples at different dilutions is presented in the table below the figure. Extracted IgG concentration as measured by human IgG-lambda ELISA (run in 2-fold dilutions) for individual samples was calculated: A = 20.384 μg/ml, B = 20.570 μg/ml, C = 30.727 μg/ml and D = 21.322 μg/ml respectively. The ELISA assay was run in duplicates in 96-well plates and mean concentration was calculated for each of the samples.

Article Snippet: Purified recombinant protein corresponding to the receptor-binding domain (RBD) of SARS-CoV-2 spike protein and derived from human cells (ProSci, Fort Collins CO) was used as antigen for immunization of mice.

Techniques: Recombinant, Enzyme-linked Immunosorbent Assay, Concentration Assay

Journal: Methods and Protocols

Article Title: SARS-CoV-2 Multi-Antigen Serology Assay

doi: 10.3390/mps4040072

Figure Lengend Snippet: Standard curves for ( a ) SARS-CoV-2 antigens and ( b ) other antigens in human plasma, such as respiratory viruses and human seasonal coronaviruses. Levels of IgG antibodies are presented as log10-transformed median fluorescence intensity (MFI) values.

Article Snippet: SARS-CoV-2 , S1 , HEK293 , The Native Antigen Company, Oxford, UK , REC31806 , SARS-CoV-2-Spike Glycoprotein (S1), recombinant antigen AA 1–674.

Techniques: Transformation Assay, Fluorescence

Journal: Methods and Protocols

Article Title: SARS-CoV-2 Multi-Antigen Serology Assay

doi: 10.3390/mps4040072

Figure Lengend Snippet: Standard curves for ( a ) SARS-CoV-2 antigens and ( b ) other antigens, such as respiratory viruses and human seasonal coronaviruses. Levels of IgM antibodies are presented as log10-transformed median fluorescence intensity (MFI) values. Solid triangles depict the negative values for each SARS-CoV-2 antigen, run at a 1:100 dilution.

Article Snippet: SARS-CoV-2 , S1 , HEK293 , The Native Antigen Company, Oxford, UK , REC31806 , SARS-CoV-2-Spike Glycoprotein (S1), recombinant antigen AA 1–674.

Techniques: Transformation Assay, Fluorescence

Journal: Methods and Protocols

Article Title: SARS-CoV-2 Multi-Antigen Serology Assay

doi: 10.3390/mps4040072

Figure Lengend Snippet: Standard curves for ( a ) SARS-CoV-2 antigens and ( b ) other antigens, such as respiratory viruses and human seasonal coronaviruses in plasma (left panels) and saliva (right panels). Levels of IgA antibodies are presented as log10-transformed median fluorescence intensity (MFI) values.

Article Snippet: SARS-CoV-2 , S1 , HEK293 , The Native Antigen Company, Oxford, UK , REC31806 , SARS-CoV-2-Spike Glycoprotein (S1), recombinant antigen AA 1–674.

Techniques: Transformation Assay, Fluorescence

Journal: Nature

Article Title: ChAdOx1 nCoV-19 vaccine prevents SARS-CoV-2 pneumonia in rhesus macaques

doi: 10.1038/s41586-020-2608-y

Figure Lengend Snippet: a. End point titer of serum IgG detected against S protein at 14 days post vaccination. No positive responses were detected in the control group. n=5, 3, 8, and 3 animals respectively examined over 1 independent experiment. b. Virus neutralizing titer in serum at 9 days post vaccination. n=3, 2, 5, and 3 animals respectively examined over 1 independent experiment. c. Summed IFN-γ ELISpot responses in splenocytes toward peptides spanning the spike protein at 14 days post vaccination. Control mice had low (<100 SFU) responses. n=5 and 8 animals respectively examined over 1 independent experiment. d. Summed frequency of spike-specific cytokine positive CD3+ T cells at 14 days post vaccination. P-value left panel: <0.0001. P-value right panel 0.0002 (IFN-γ−IL-4); 0.0001 (IFN-γ-IL-10); 0.0054 (TNF-α-IL-4); 0.0022 (TNF-α-IL-10). n=5 and 8 animals respectively examined over 1 independent experiment. BALB/c = red; CD1 = blue; vaccinated = circle; control = square; dotted line = limit of detection; line = mean; SFU = spot-forming units; Spl. = splenocytes; * = p<0.05. Statistical significance determined via 2-way ANOVA (repeated measure) and post-hoc positive test.

Article Snippet: MaxiSorp plates (Nunc) were coated with S1 (monomeric, AA 1–674) or S2 (monomeric, AA 685–1211 The Native Antigen Company; 50 ng/well) or with prefusion stabilized SARS-CoV-2 spike protein (250 ng/well) in PBS for overnight adsorption at 4°C.

Techniques: Virus, Enzyme-linked Immunospot

Journal: Nature

Article Title: ChAdOx1 nCoV-19 vaccine prevents SARS-CoV-2 pneumonia in rhesus macaques

doi: 10.1038/s41586-020-2608-y

Figure Lengend Snippet: No histological changes were observed in the lungs of ChAdOx1 nCoV-19 prime (a) and prime-boost (b) vaccinated animals. c) Interstitial pneumonia with edema (asterisk), type II pneumocyte hyperplasia (arrowhead) and syncytial cells (arrow) in control animals. No SARS-CoV-2 antigen was detected by immunohistochemistry in the lungs of ChAdOx1 nCoV-19 prime (d) and prime-boost (e) vaccinated animals. f) SARS-CoV-2 antigen (visible as red-brown staining) was detected by immunohistochemistry in type I and type II pneumocytes in the lungs of control animals. 18 sections, taken from 6 different lung lobes are evaluated for each animal; a representative lesion from each group was selected for the figure. Magnification: 400x, scale bar = 20 µm.

Article Snippet: MaxiSorp plates (Nunc) were coated with S1 (monomeric, AA 1–674) or S2 (monomeric, AA 685–1211 The Native Antigen Company; 50 ng/well) or with prefusion stabilized SARS-CoV-2 spike protein (250 ng/well) in PBS for overnight adsorption at 4°C.

Techniques: Immunohistochemistry, Staining

Journal: Nature

Article Title: ChAdOx1 nCoV-19 vaccine prevents SARS-CoV-2 pneumonia in rhesus macaques

doi: 10.1038/s41586-020-2608-y

Figure Lengend Snippet: Viral load in lung tissue of rhesus macaques challenged with SARS-CoV-2 at 7 DPI.

Article Snippet: MaxiSorp plates (Nunc) were coated with S1 (monomeric, AA 1–674) or S2 (monomeric, AA 685–1211 The Native Antigen Company; 50 ng/well) or with prefusion stabilized SARS-CoV-2 spike protein (250 ng/well) in PBS for overnight adsorption at 4°C.

Techniques: