Journal: Scientific Reports

Article Title: Immunogenicity and protection efficacy of a COVID-19 DNA vaccine encoding spike protein with D614G mutation and optimization of large-scale DNA vaccine production

doi: 10.1038/s41598-024-64690-5

Figure Lengend Snippet: In silico analyses of vaccine antigen. ( A ) Variations detected in Spike gene of 20 SARS-CoV-2 isolates collected from various provinces of Türkiye. The map in figure was generated by Microsoft 365 Excel. ( B ) A1841G mutation causes D614G amino acid alteration ( C ) 3D image of human ACE2 receptor docked with Wuhan Spike protein and coSpikeD614G vaccine antigen. Purple color: Spike protein; blue color: receptor binding domain (RBD) of Spike protein; green color: human ACE2 receptor; red color: receptor binding motif of human ACE2 protein.

Article Snippet: Neutralizing antibody responses in vaccinated mice sera were analyzed using an ELISA-based SARS-CoV-2 surrogate Virus Neutralization Test (sVNT) which is a S1 protein/ACE2 ligand binding assay) (AffinityImmuno, Canada) according to the manufacturer’s protocol.

Techniques: In Silico, Generated, Mutagenesis, Binding Assay

Journal: Scientific Reports

Article Title: Immunogenicity and protection efficacy of a COVID-19 DNA vaccine encoding spike protein with D614G mutation and optimization of large-scale DNA vaccine production

doi: 10.1038/s41598-024-64690-5

Figure Lengend Snippet: Animal studies and humoral immune response elicited by pcoSpikeD614G DNA vaccine. ( A ) Brief timeline of animal studies. BALB/c mice were immunized to assess the immunogenicity and K18-hACE2 transgenic mice were immunized to determine protective efficacy conferred by pcoSpikeD614G DNA vaccine ( B ) Western blot image shows the presence of anti-S1 and anti S1 + S2 antibody responses in sera of vaccinated BALB/c mice. Lane 1: Protein ladder; Lanes 2 and 3: recombinant S1 protein probed with pooled pcoSpikeD614G vaccinated mice sera immunized thrice through ID + EP and IM routes. Red arrowheads show the recombinant S1 proteins with a size of over \documentclass[12pt]{minimal} \usepackage{amsmath} \usepackage{wasysym} \usepackage{amsfonts} \usepackage{amssymb} \usepackage{amsbsy} \usepackage{mathrsfs} \usepackage{upgreek} \setlength{\oddsidemargin}{-69pt} \begin{document}$$\sim$$\end{document} ∼ 76.41 kDa due to glycosylation; Lanes 4 and 5: recombinant S1 + S2 protein pooled with pcoSpikeD614G vaccinated mice sera immunized thrice through ID + EP and IM routes. Blue arrowheads show the recombinant S1 + S2 proteins with a size of over \documentclass[12pt]{minimal} \usepackage{amsmath} \usepackage{wasysym} \usepackage{amsfonts} \usepackage{amssymb} \usepackage{amsbsy} \usepackage{mathrsfs} \usepackage{upgreek} \setlength{\oddsidemargin}{-69pt} \begin{document}$$\sim$$\end{document} ∼ 138.5 kDa due to glycosylation; Lane 6–7 and 8–9: recombinant S1 and S1 + S2 proteins probed with pooled vaccinated mice sera administered with empty pVAX1. Original Western blot images have been shown in Supplementary file named S5 Figure ( C ) The anti-S1 IgG kinetics derived from mice immunized with pcoSpikeD614G administered through ID + EP and IM routes show significant increase at day 70 compared to controls ( D ) IgG2a/IgG1 polarization was assessed at days 0 and 70 for empty pVAX1 and pcoSpikeD614G and IgG2a responses were significantly higher ( E ) The ratio of IgG2a/IgG1 is slightly higher in pcoSpikeD614G administered through IM route ( F ) SARS-CoV-2 Surrogate Virus Neutralization Test showed that inhibition potential of 1:10 diluted sera obtained from vaccinated mice at day 70 was significantly higher than controls ( G ) SARS-CoV-2 50% neutralization titers (VNT50) in sera of in pcoSpikeD614G immunized mice collated on day 70. Data are presented as GMT ± SD.

Article Snippet: Neutralizing antibody responses in vaccinated mice sera were analyzed using an ELISA-based SARS-CoV-2 surrogate Virus Neutralization Test (sVNT) which is a S1 protein/ACE2 ligand binding assay) (AffinityImmuno, Canada) according to the manufacturer’s protocol.

Techniques: Transgenic Assay, Western Blot, Recombinant, Derivative Assay, Virus, Neutralization, Inhibition

Journal: Scientific Reports

Article Title: Immunogenicity and protection efficacy of a COVID-19 DNA vaccine encoding spike protein with D614G mutation and optimization of large-scale DNA vaccine production

doi: 10.1038/s41598-024-64690-5

Figure Lengend Snippet: Protection conferred by pcoSpikeD614G DNA vaccine in K18-hACE2 transgenic mice challenged by SARS-CoV-2. ( A ) Gross pathological scoring of lungs to evaluate the level of pneumonia. Healthy lungs were scored as 0, edema-hyperemia between 0.5 and 1, pneumonia lesions were between 1.5 and 5 and dead mice were scored as 5. ( B ) Mean Ct values of RT-qPCR targeting NC gene of SARS‐CoV‐2 derived from the lungs of mice. The virus load is represented by Crossing point threshold (Ct) values. The Ct value of a negative PCR sample is accepted as 40 ( C ) Histopathology scoring of lungs to evaluate the level of inflammation status. Absence of inflammation is represented by 0 and various level of inflammation were scored between 1 and 3. Three lungs from the control group and one lung from the mice group immunized with pcoSpikeD614G administered through IM route were not scored ( D, E ) In the group of mice immunized with pcoSpikeD614G administered through IM route, images show normal lungs at magnifications of 4 × and 10 × . Arrow shows alveoli with normal morphology ( F, G ) In the group of mice immunized with pcoSpikeD614G administered through ID + EP route, images show normal lungs at magnifications of 4 × and 10 × . Arrow shows alveoli with normal morphology. ( H, I ) In the control group of mice, images show lungs with interstitial inflammatory cell infiltration at magnifications of 4 × and 10 × . Asterisks indicate areas of lymphocyte infiltration in the interalveolar spaces. ( J ) Kaplan–Meier Survival analysis after intranasal instillation of 10 5 TCID 50 virus for 3 consecutive days.

Article Snippet: Neutralizing antibody responses in vaccinated mice sera were analyzed using an ELISA-based SARS-CoV-2 surrogate Virus Neutralization Test (sVNT) which is a S1 protein/ACE2 ligand binding assay) (AffinityImmuno, Canada) according to the manufacturer’s protocol.

Techniques: Transgenic Assay, Quantitative RT-PCR, Derivative Assay, Virus, Histopathology

Journal: medRxiv

Article Title: Long-term immunogenicity of BNT162b2 vaccination in the elderly and in younger health care workers

doi: 10.1101/2021.08.26.21262468

Figure Lengend Snippet: (A) Anti-S1 IgG in serum measured by a microarray-based immunoassay, (B) serum pseudovirus neutralization against the Delta VOC two and six months after vaccination measured by pNT, and (C) SARS-CoV-2 S1 specific T cell response detected by IGRA. Dotted horizontal lines indicate themanufacturer’s threshold for anti-S1 IgG ≥1 S/Co (A) and the lower limit of detection (1:10 dilution) for pNT (B). Horizontal lines within plotted data regions indicate the median and interquartile range, except for pNT, where the geometric mean and 95% confidence interval is shown. P values (all <0.0001) were calculated by non-parametric Mann Whitney U test. S/Co: signal-to-cutoff, pNT: pseudovirus neutralization test, ID 50 : 50% inhibition dilution, IGRA: interferon-γ release assay, IU: international units.

Article Snippet: Functional neutralization capacity was investigated using a commercially available ELISA-based SARS-CoV-2 RBD-ACE2 binding inhibition assay (surrogate SARS-CoV-2 neutralization test (sVNT) cPass (medac GmbH, https://international.medac.de ).

Techniques: Microarray, Neutralization, MANN-WHITNEY, Inhibition, Release Assay

Journal: medRxiv

Article Title: Long-term immunogenicity of BNT162b2 vaccination in the elderly and in younger health care workers

doi: 10.1101/2021.08.26.21262468

Figure Lengend Snippet: ( A ) Anti-SARS-CoV-2 N, ( B ) RBD- ( C ) and full-spike IgG measured in the serum of BNT162b2 vaccinated HCW and elderly persons six months after the first vaccination. ( D ) Neutralizing capacity was measured by sVNT and ( E ) serum neutralization against Alpha (B.1.1.7) VOC detected by pNT in vaccinated HCW and elderly persons six months after the first vaccination. ( F ) Binding capacity of serum IgG against six different RBDs of SARS-CoV-2 variants carrying the indicated mutations in HCW and elderly, measured by ELISA. Dotted lines indicate the manufacturer’s threshold values: for anti-N, anti-RBD, and anti-full spike IgG ≥1 S/Co, for sVNT >30%, and the lower limit of detection (1:10 dilution) for pNT. Lines indicate the median and interquartile range except for pNT, where the geometric mean and 95% confidence interval are shown. P values were calculated by the non-parametric Mann Whitney U test or Kruskal-Wallis test with Dunn’s multiple comparisons test. S/Co: signal-to-cutoff, N: nucleocapsid protein, RBD: receptor-binding domain, sVNT: surrogate virus neutralization test, ACE2: angiotensin-converting enzyme 2, ID50: 50% inhibition dilution.

Article Snippet: Functional neutralization capacity was investigated using a commercially available ELISA-based SARS-CoV-2 RBD-ACE2 binding inhibition assay (surrogate SARS-CoV-2 neutralization test (sVNT) cPass (medac GmbH, https://international.medac.de ).

Techniques: Neutralization, Binding Assay, Enzyme-linked Immunosorbent Assay, MANN-WHITNEY, Inhibition