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mouse anti sars cov 2 spike monoclonal antibody  (Sino Biological)


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    Structured Review

    Sino Biological mouse anti sars cov 2 spike monoclonal antibody
    IN <t>Ad4-SARS-CoV-2</t> induces binding antibodies on the nasal mucosa Heatmap depicting binding IgA (left), IgG (center) and IgM (right) responses observed in hamster nasal wash (top), BAL (center), and serum (bottom) for each immunization group (columns) against the indicated SARS-CoV antigens (rows), including Spike (S), the S2 and receptor-binding domain (RBD), and Nucleocapsid protein. Mean response values in each group of 6 animals are graphed.
    Mouse Anti Sars Cov 2 Spike Monoclonal Antibody, supplied by Sino Biological, used in various techniques. Bioz Stars score: 96/100, based on 171 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/mouse+anti+spike+antibody/pmc13090634-374-19-24?v=Sino+Biological
    Average 96 stars, based on 171 article reviews
    mouse anti sars cov 2 spike monoclonal antibody - by Bioz Stars, 2026-07
    96/100 stars

    Images

    1) Product Images from "Intranasal replicating adenovirus type 4-vectored SARS-CoV-2 vaccines induce durable and efficacious responses in preclinical testing"

    Article Title: Intranasal replicating adenovirus type 4-vectored SARS-CoV-2 vaccines induce durable and efficacious responses in preclinical testing

    Journal: iScience

    doi: 10.1016/j.isci.2026.115433

    IN Ad4-SARS-CoV-2 induces binding antibodies on the nasal mucosa Heatmap depicting binding IgA (left), IgG (center) and IgM (right) responses observed in hamster nasal wash (top), BAL (center), and serum (bottom) for each immunization group (columns) against the indicated SARS-CoV antigens (rows), including Spike (S), the S2 and receptor-binding domain (RBD), and Nucleocapsid protein. Mean response values in each group of 6 animals are graphed.
    Figure Legend Snippet: IN Ad4-SARS-CoV-2 induces binding antibodies on the nasal mucosa Heatmap depicting binding IgA (left), IgG (center) and IgM (right) responses observed in hamster nasal wash (top), BAL (center), and serum (bottom) for each immunization group (columns) against the indicated SARS-CoV antigens (rows), including Spike (S), the S2 and receptor-binding domain (RBD), and Nucleocapsid protein. Mean response values in each group of 6 animals are graphed.

    Techniques Used: Binding Assay

    IN Ad4-Spike immunization limits weight loss following SARS-CoV-2 challenge Average percent body weight relative to the day of challenge, at either 56 days or 174 days following the first immunization. Hamsters were weighed once individually per day. Dots represent median and bars represent standard error for each group ( N = 10 hamsters).
    Figure Legend Snippet: IN Ad4-Spike immunization limits weight loss following SARS-CoV-2 challenge Average percent body weight relative to the day of challenge, at either 56 days or 174 days following the first immunization. Hamsters were weighed once individually per day. Dots represent median and bars represent standard error for each group ( N = 10 hamsters).

    Techniques Used:

    IN immunization with Ad4-Anc reduces lung pathology following SARS-CoV-2 challenge Hamsters from each group were sacrificed four days after challenge (day 60 or 178) to assess disease severity in the lungs. Scores ranging from 1 to 5 were assigned based on pneumonia severity and IHC staining in the bronchial epithelium and alveoli. Average scores for each group ( N = 4 hamsters), and the percent of the lung affected by pneumonia, are indicated.
    Figure Legend Snippet: IN immunization with Ad4-Anc reduces lung pathology following SARS-CoV-2 challenge Hamsters from each group were sacrificed four days after challenge (day 60 or 178) to assess disease severity in the lungs. Scores ranging from 1 to 5 were assigned based on pneumonia severity and IHC staining in the bronchial epithelium and alveoli. Average scores for each group ( N = 4 hamsters), and the percent of the lung affected by pneumonia, are indicated.

    Techniques Used: Immunohistochemistry

    IN Ad4-Spike induces the durable restriction of SARS-CoV-2 replication after transmission. Weight loss and viral replication data following transmission at day 268 (A) Percent body weight relative to the day of challenge, represented as in . Dots represent the median for each group ( N = 6 hamsters) and bars represent standard error (B) AUC of viral copies measured in oropharyngeal swabs of individual hamsters, as represented in . Values for each hamster are indicated with dots, the median AUC value for each group ( N = 6 hamsters) is represented as a black line. The dotted line indicates the AUC limit of detection over 14 days (12,600 copies). (C) Curves representing viral replication over time after challenge. The limit of detection is depicted by the dotted line (10 copies per m L). Dots represent medians and bars represent standard error for each group ( N = 6 animals). The Wilcoxon rank-sum test was used for pairwise comparisons to generate the indicated p values.
    Figure Legend Snippet: IN Ad4-Spike induces the durable restriction of SARS-CoV-2 replication after transmission. Weight loss and viral replication data following transmission at day 268 (A) Percent body weight relative to the day of challenge, represented as in . Dots represent the median for each group ( N = 6 hamsters) and bars represent standard error (B) AUC of viral copies measured in oropharyngeal swabs of individual hamsters, as represented in . Values for each hamster are indicated with dots, the median AUC value for each group ( N = 6 hamsters) is represented as a black line. The dotted line indicates the AUC limit of detection over 14 days (12,600 copies). (C) Curves representing viral replication over time after challenge. The limit of detection is depicted by the dotted line (10 copies per m L). Dots represent medians and bars represent standard error for each group ( N = 6 animals). The Wilcoxon rank-sum test was used for pairwise comparisons to generate the indicated p values.

    Techniques Used: Transmission Assay



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    IN <t>Ad4-SARS-CoV-2</t> induces binding antibodies on the nasal mucosa Heatmap depicting binding IgA (left), IgG (center) and IgM (right) responses observed in hamster nasal wash (top), BAL (center), and serum (bottom) for each immunization group (columns) against the indicated SARS-CoV antigens (rows), including Spike (S), the S2 and receptor-binding domain (RBD), and Nucleocapsid protein. Mean response values in each group of 6 animals are graphed.
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    Image Search Results


    IN Ad4-SARS-CoV-2 induces binding antibodies on the nasal mucosa Heatmap depicting binding IgA (left), IgG (center) and IgM (right) responses observed in hamster nasal wash (top), BAL (center), and serum (bottom) for each immunization group (columns) against the indicated SARS-CoV antigens (rows), including Spike (S), the S2 and receptor-binding domain (RBD), and Nucleocapsid protein. Mean response values in each group of 6 animals are graphed.

    Journal: iScience

    Article Title: Intranasal replicating adenovirus type 4-vectored SARS-CoV-2 vaccines induce durable and efficacious responses in preclinical testing

    doi: 10.1016/j.isci.2026.115433

    Figure Lengend Snippet: IN Ad4-SARS-CoV-2 induces binding antibodies on the nasal mucosa Heatmap depicting binding IgA (left), IgG (center) and IgM (right) responses observed in hamster nasal wash (top), BAL (center), and serum (bottom) for each immunization group (columns) against the indicated SARS-CoV antigens (rows), including Spike (S), the S2 and receptor-binding domain (RBD), and Nucleocapsid protein. Mean response values in each group of 6 animals are graphed.

    Article Snippet: For surface staining cells were harvested using 0.01 M EDTA in PBS 48 h after infection and stained with mouse anti-SARS-CoV-2 spike monoclonal antibody (Sino Biological, Inc., Beijing, China) (50 μL at 1.0 μg/mL final concentration), washed twice with phosphate buffered saline (PBS) (Quality Biological, Gaithersburg, MD, USA) containing 1% bovine serum albumin (Sigma-Aldrich, St. Louis, MO, USA), then stained with a goat anti-mouse IgG PE-conjugated secondary antibody (BioLegend, San Diego, CA, USA) (100 μL at 2.0 μg/mL final concentration) at 37°C for 30 min each.

    Techniques: Binding Assay

    IN Ad4-Spike immunization limits weight loss following SARS-CoV-2 challenge Average percent body weight relative to the day of challenge, at either 56 days or 174 days following the first immunization. Hamsters were weighed once individually per day. Dots represent median and bars represent standard error for each group ( N = 10 hamsters).

    Journal: iScience

    Article Title: Intranasal replicating adenovirus type 4-vectored SARS-CoV-2 vaccines induce durable and efficacious responses in preclinical testing

    doi: 10.1016/j.isci.2026.115433

    Figure Lengend Snippet: IN Ad4-Spike immunization limits weight loss following SARS-CoV-2 challenge Average percent body weight relative to the day of challenge, at either 56 days or 174 days following the first immunization. Hamsters were weighed once individually per day. Dots represent median and bars represent standard error for each group ( N = 10 hamsters).

    Article Snippet: For surface staining cells were harvested using 0.01 M EDTA in PBS 48 h after infection and stained with mouse anti-SARS-CoV-2 spike monoclonal antibody (Sino Biological, Inc., Beijing, China) (50 μL at 1.0 μg/mL final concentration), washed twice with phosphate buffered saline (PBS) (Quality Biological, Gaithersburg, MD, USA) containing 1% bovine serum albumin (Sigma-Aldrich, St. Louis, MO, USA), then stained with a goat anti-mouse IgG PE-conjugated secondary antibody (BioLegend, San Diego, CA, USA) (100 μL at 2.0 μg/mL final concentration) at 37°C for 30 min each.

    Techniques:

    IN immunization with Ad4-Anc reduces lung pathology following SARS-CoV-2 challenge Hamsters from each group were sacrificed four days after challenge (day 60 or 178) to assess disease severity in the lungs. Scores ranging from 1 to 5 were assigned based on pneumonia severity and IHC staining in the bronchial epithelium and alveoli. Average scores for each group ( N = 4 hamsters), and the percent of the lung affected by pneumonia, are indicated.

    Journal: iScience

    Article Title: Intranasal replicating adenovirus type 4-vectored SARS-CoV-2 vaccines induce durable and efficacious responses in preclinical testing

    doi: 10.1016/j.isci.2026.115433

    Figure Lengend Snippet: IN immunization with Ad4-Anc reduces lung pathology following SARS-CoV-2 challenge Hamsters from each group were sacrificed four days after challenge (day 60 or 178) to assess disease severity in the lungs. Scores ranging from 1 to 5 were assigned based on pneumonia severity and IHC staining in the bronchial epithelium and alveoli. Average scores for each group ( N = 4 hamsters), and the percent of the lung affected by pneumonia, are indicated.

    Article Snippet: For surface staining cells were harvested using 0.01 M EDTA in PBS 48 h after infection and stained with mouse anti-SARS-CoV-2 spike monoclonal antibody (Sino Biological, Inc., Beijing, China) (50 μL at 1.0 μg/mL final concentration), washed twice with phosphate buffered saline (PBS) (Quality Biological, Gaithersburg, MD, USA) containing 1% bovine serum albumin (Sigma-Aldrich, St. Louis, MO, USA), then stained with a goat anti-mouse IgG PE-conjugated secondary antibody (BioLegend, San Diego, CA, USA) (100 μL at 2.0 μg/mL final concentration) at 37°C for 30 min each.

    Techniques: Immunohistochemistry

    IN Ad4-Spike induces the durable restriction of SARS-CoV-2 replication after transmission. Weight loss and viral replication data following transmission at day 268 (A) Percent body weight relative to the day of challenge, represented as in . Dots represent the median for each group ( N = 6 hamsters) and bars represent standard error (B) AUC of viral copies measured in oropharyngeal swabs of individual hamsters, as represented in . Values for each hamster are indicated with dots, the median AUC value for each group ( N = 6 hamsters) is represented as a black line. The dotted line indicates the AUC limit of detection over 14 days (12,600 copies). (C) Curves representing viral replication over time after challenge. The limit of detection is depicted by the dotted line (10 copies per m L). Dots represent medians and bars represent standard error for each group ( N = 6 animals). The Wilcoxon rank-sum test was used for pairwise comparisons to generate the indicated p values.

    Journal: iScience

    Article Title: Intranasal replicating adenovirus type 4-vectored SARS-CoV-2 vaccines induce durable and efficacious responses in preclinical testing

    doi: 10.1016/j.isci.2026.115433

    Figure Lengend Snippet: IN Ad4-Spike induces the durable restriction of SARS-CoV-2 replication after transmission. Weight loss and viral replication data following transmission at day 268 (A) Percent body weight relative to the day of challenge, represented as in . Dots represent the median for each group ( N = 6 hamsters) and bars represent standard error (B) AUC of viral copies measured in oropharyngeal swabs of individual hamsters, as represented in . Values for each hamster are indicated with dots, the median AUC value for each group ( N = 6 hamsters) is represented as a black line. The dotted line indicates the AUC limit of detection over 14 days (12,600 copies). (C) Curves representing viral replication over time after challenge. The limit of detection is depicted by the dotted line (10 copies per m L). Dots represent medians and bars represent standard error for each group ( N = 6 animals). The Wilcoxon rank-sum test was used for pairwise comparisons to generate the indicated p values.

    Article Snippet: For surface staining cells were harvested using 0.01 M EDTA in PBS 48 h after infection and stained with mouse anti-SARS-CoV-2 spike monoclonal antibody (Sino Biological, Inc., Beijing, China) (50 μL at 1.0 μg/mL final concentration), washed twice with phosphate buffered saline (PBS) (Quality Biological, Gaithersburg, MD, USA) containing 1% bovine serum albumin (Sigma-Aldrich, St. Louis, MO, USA), then stained with a goat anti-mouse IgG PE-conjugated secondary antibody (BioLegend, San Diego, CA, USA) (100 μL at 2.0 μg/mL final concentration) at 37°C for 30 min each.

    Techniques: Transmission Assay

    PAD expression in vivo and in vitro after SARS-CoV-2 infection (A) Volcano plot relative to the reanalysis of transcriptomic data from ten studies of healthy lungs and four studies of patients with COVID-19 (Delorey et al.). (B and C) Calu3 cells are infected with 2020B.1 (MOI 0.01), Delta (MOI 0.01), or Omicron (MOI 0.01) SARS-CoV-2 variants, and PAD4 (B) and PAD2 (C) mRNA are assessed by RT-qPCR. Data are normalized to the housekeeping gene PGK1 and expressed as mean fold change ±SEM over mock-infected cells of three independent experiments. ∗ p < 0.05, ∗∗ p < 0.01, and ∗∗∗ p < 0.001; one-way ANOVA followed by Bonferroni’s post-test. (D) Protein lysates from Calu3 infected with SARS-CoV-2 (2020B.1 variant, MOI 0.1 PFU/cell) or uninfected cells (mock) at different hours post-infection (hpi) are analyzed for citrullinated proteins using an Rh-PG citrulline-specific probe (left panel) or an anti-CCP antibody (right panel). Samples are subjected to gel electrophoresis, and SARS-CoV-2 infection is confirmed using an anti-SARS-CoV-2 Spike (S SARS-CoV-2) antibody. β-actin is used as a loading control. One representative blot of three independent experiments is shown. (E) Volcano plot depicts host (gray dots) and viral (red dots) citrullinated proteins of SARS-infected vs. mock-infected cells at 48 hpi (2020B.1 variant, MOI 0.1 PFU/cell). The x axis represents the ratio of citrullination between mock and infected cells at the indicated time points, while the y axis indicates the statistical significance. Both variables are plotted on a logarithmic scale ( n = 3). (F) PANTHER classification of over-citrullinated cellular proteins based on protein classes.

    Journal: iScience

    Article Title: Targeting peptidyl-arginine deiminase 4 suppresses SARS-CoV-2 replication and modulates the inflammatory response

    doi: 10.1016/j.isci.2026.115038

    Figure Lengend Snippet: PAD expression in vivo and in vitro after SARS-CoV-2 infection (A) Volcano plot relative to the reanalysis of transcriptomic data from ten studies of healthy lungs and four studies of patients with COVID-19 (Delorey et al.). (B and C) Calu3 cells are infected with 2020B.1 (MOI 0.01), Delta (MOI 0.01), or Omicron (MOI 0.01) SARS-CoV-2 variants, and PAD4 (B) and PAD2 (C) mRNA are assessed by RT-qPCR. Data are normalized to the housekeeping gene PGK1 and expressed as mean fold change ±SEM over mock-infected cells of three independent experiments. ∗ p < 0.05, ∗∗ p < 0.01, and ∗∗∗ p < 0.001; one-way ANOVA followed by Bonferroni’s post-test. (D) Protein lysates from Calu3 infected with SARS-CoV-2 (2020B.1 variant, MOI 0.1 PFU/cell) or uninfected cells (mock) at different hours post-infection (hpi) are analyzed for citrullinated proteins using an Rh-PG citrulline-specific probe (left panel) or an anti-CCP antibody (right panel). Samples are subjected to gel electrophoresis, and SARS-CoV-2 infection is confirmed using an anti-SARS-CoV-2 Spike (S SARS-CoV-2) antibody. β-actin is used as a loading control. One representative blot of three independent experiments is shown. (E) Volcano plot depicts host (gray dots) and viral (red dots) citrullinated proteins of SARS-infected vs. mock-infected cells at 48 hpi (2020B.1 variant, MOI 0.1 PFU/cell). The x axis represents the ratio of citrullination between mock and infected cells at the indicated time points, while the y axis indicates the statistical significance. Both variables are plotted on a logarithmic scale ( n = 3). (F) PANTHER classification of over-citrullinated cellular proteins based on protein classes.

    Article Snippet: The following primary antibodies were sequentially applied to the slides: SARS-CoV-1/2 Spike Protein (clone 2B3E5, Cell Signaling), mouse anti-mouse β-tubulin IV (clone T7941, Merck), rat anti-mouse F4/80 (clone CI:A3-1, Bio-Rad Laboratories), rabbit anti-mouse PAD2 (polyclonal, Proteintech), rabbit anti-mouse PAD4 (polyclonal, Proteintech), rabbit anti-mouse Ly6g (clone EPR22909-135, Abcam), rat anti-mouse CD3 (clone CD3-12, Abcam), and rabbit anti-mouse cytokeratin 8 (clone EP1628Y, Abcam).

    Techniques: Expressing, In Vivo, In Vitro, Infection, Quantitative RT-PCR, Variant Assay, Nucleic Acid Electrophoresis, Control

    SARS-CoV-2 infection affects protein citrullination by modulating PAD expression in vivo (A and B) PAD4 and PAD2 mRNA expression in mouse lungs (A) and brains (B) assessed by RT-qPCR. Each dot represents an individual mouse sample ( n = 6). Data are normalized to the housekeeping gene β-actin and presented relative to one randomly selected non-infected sample. Data represent mean ± SEM. Statistical significance is determined using non parametric t test, ∗ p < 0.05. (C) Western blot analysis of protein lysates from pooled uninfected (mock) and SARS-CoV-2-infected (6 dpi, Delta variant) mouse lungs and brains. The analysis is performed using antibodies against PAD2, PAD4, and β-actin, the latter to ensure equal loading. A representative blot from three independent experiments is shown. (D) Multiplex immunofluorescence staining for the detection of SARS-CoV-2-targeted cells in mouse brains and lung sections at 6 dpi and mock control. S SARS-CoV-2 protein, PAD2, and PAD4 positive signals are represented in green, pink, and yellow, respectively. Cell nuclei are visualized by DAPI (blue). Original magnification 20×. (E) Quantification of PAD2-and PAD4-positive cells is performed using the inForm Image Analysis software (Akoya Biosciences). For each mouse ( n = 6), one representative section is analyzed to determine cell density (cells/mm 2 ). Data represent mean ± SEM. Statistical significance is determined using non parametric t test, ∗ p < 0.05. (F) Volcano plot shows citrullinated proteins in pooled protein lysates from SARS-CoV-2-infected vs. mock-infected mouse lungs at 6 dpi. Lysates are incubated with biotin-PG to isolate citrullinated proteins on streptavidin agarose, followed by on-bead tryptic digestion and LC-MS/MS analysis. Each dot represents an identified citrullinated protein. The x axis shows the citrullination ratio between mock and infected samples, and the y axis indicates statistical significance, both on a logarithmic scale ( n = 3). (G) PANTHER classification of over-citrullinated cellular proteins based on protein classes.

    Journal: iScience

    Article Title: Targeting peptidyl-arginine deiminase 4 suppresses SARS-CoV-2 replication and modulates the inflammatory response

    doi: 10.1016/j.isci.2026.115038

    Figure Lengend Snippet: SARS-CoV-2 infection affects protein citrullination by modulating PAD expression in vivo (A and B) PAD4 and PAD2 mRNA expression in mouse lungs (A) and brains (B) assessed by RT-qPCR. Each dot represents an individual mouse sample ( n = 6). Data are normalized to the housekeeping gene β-actin and presented relative to one randomly selected non-infected sample. Data represent mean ± SEM. Statistical significance is determined using non parametric t test, ∗ p < 0.05. (C) Western blot analysis of protein lysates from pooled uninfected (mock) and SARS-CoV-2-infected (6 dpi, Delta variant) mouse lungs and brains. The analysis is performed using antibodies against PAD2, PAD4, and β-actin, the latter to ensure equal loading. A representative blot from three independent experiments is shown. (D) Multiplex immunofluorescence staining for the detection of SARS-CoV-2-targeted cells in mouse brains and lung sections at 6 dpi and mock control. S SARS-CoV-2 protein, PAD2, and PAD4 positive signals are represented in green, pink, and yellow, respectively. Cell nuclei are visualized by DAPI (blue). Original magnification 20×. (E) Quantification of PAD2-and PAD4-positive cells is performed using the inForm Image Analysis software (Akoya Biosciences). For each mouse ( n = 6), one representative section is analyzed to determine cell density (cells/mm 2 ). Data represent mean ± SEM. Statistical significance is determined using non parametric t test, ∗ p < 0.05. (F) Volcano plot shows citrullinated proteins in pooled protein lysates from SARS-CoV-2-infected vs. mock-infected mouse lungs at 6 dpi. Lysates are incubated with biotin-PG to isolate citrullinated proteins on streptavidin agarose, followed by on-bead tryptic digestion and LC-MS/MS analysis. Each dot represents an identified citrullinated protein. The x axis shows the citrullination ratio between mock and infected samples, and the y axis indicates statistical significance, both on a logarithmic scale ( n = 3). (G) PANTHER classification of over-citrullinated cellular proteins based on protein classes.

    Article Snippet: The following primary antibodies were sequentially applied to the slides: SARS-CoV-1/2 Spike Protein (clone 2B3E5, Cell Signaling), mouse anti-mouse β-tubulin IV (clone T7941, Merck), rat anti-mouse F4/80 (clone CI:A3-1, Bio-Rad Laboratories), rabbit anti-mouse PAD2 (polyclonal, Proteintech), rabbit anti-mouse PAD4 (polyclonal, Proteintech), rabbit anti-mouse Ly6g (clone EPR22909-135, Abcam), rat anti-mouse CD3 (clone CD3-12, Abcam), and rabbit anti-mouse cytokeratin 8 (clone EP1628Y, Abcam).

    Techniques: Infection, Expressing, In Vivo, Quantitative RT-PCR, Western Blot, Variant Assay, Multiplex Assay, Immunofluorescence, Staining, Control, Software, Incubation, Liquid Chromatography with Mass Spectroscopy

    Antiviral effects of PAD inhibitors against SARS-CoV-2 in vitro (A and B) Quantification of SARS-CoV-2 2020B.1 (MOI 0.1) viral particle production in Calu-3 and Huh7.5 cells (at 24 hpi and 48 hpi, respectively) treated with the increasing concentrations of BB-Cl (A) or GSK199 (B), determined by plaque assay. Results are expressed as a percentage relative to vehicle-treated cells (DMSO, marked as 0 on the graph). Data are presented as means ± SEM from four independent experiments and are analyzed by one-way ANOVA followed by Bonferroni’s post-test. ∗ p < 0.05, ∗∗ p < 0.01, and ∗∗∗ p < 0.001. (C and D) Cell viability of uninfected Calu3 and Huh7.5 cells treated with the indicated concentrations of BB-Cl (C) and GSK199 (D) for 72 h, determined for each concentration by MTT assay. Values are expressed as means ± SEM of three independent experiments. (E) Viral titers of different SARS-CoV-2 variants are measured in Calu-3 cells at 24 hpi (MOI = 0.1) following treatment with BB-Cl (5 μM) or GSK199 (20 μM). Viral particle production is determined by plaque assay. Data are expressed as mean ± SEM from four independent experiments and analyzed by t test. ∗ p < 0.05, ∗∗ p < 0.01, and ∗∗∗ p < 0.001. (F) Viral titers of different SARS-CoV-2 variants (as indicated in the figure) are measured in Huh7.5 cells at 48 hpi (MOI = 0.1) following treatment with BB-Cl (5 μM) or GSK199 (20 μM). Viral particle production is determined by plaque assay. Data are expressed as mean ± SEM from four independent experiments and analyzed by t test. ∗ p < 0.05, ∗∗ p < 0.01, and ∗∗∗ p < 0.001.

    Journal: iScience

    Article Title: Targeting peptidyl-arginine deiminase 4 suppresses SARS-CoV-2 replication and modulates the inflammatory response

    doi: 10.1016/j.isci.2026.115038

    Figure Lengend Snippet: Antiviral effects of PAD inhibitors against SARS-CoV-2 in vitro (A and B) Quantification of SARS-CoV-2 2020B.1 (MOI 0.1) viral particle production in Calu-3 and Huh7.5 cells (at 24 hpi and 48 hpi, respectively) treated with the increasing concentrations of BB-Cl (A) or GSK199 (B), determined by plaque assay. Results are expressed as a percentage relative to vehicle-treated cells (DMSO, marked as 0 on the graph). Data are presented as means ± SEM from four independent experiments and are analyzed by one-way ANOVA followed by Bonferroni’s post-test. ∗ p < 0.05, ∗∗ p < 0.01, and ∗∗∗ p < 0.001. (C and D) Cell viability of uninfected Calu3 and Huh7.5 cells treated with the indicated concentrations of BB-Cl (C) and GSK199 (D) for 72 h, determined for each concentration by MTT assay. Values are expressed as means ± SEM of three independent experiments. (E) Viral titers of different SARS-CoV-2 variants are measured in Calu-3 cells at 24 hpi (MOI = 0.1) following treatment with BB-Cl (5 μM) or GSK199 (20 μM). Viral particle production is determined by plaque assay. Data are expressed as mean ± SEM from four independent experiments and analyzed by t test. ∗ p < 0.05, ∗∗ p < 0.01, and ∗∗∗ p < 0.001. (F) Viral titers of different SARS-CoV-2 variants (as indicated in the figure) are measured in Huh7.5 cells at 48 hpi (MOI = 0.1) following treatment with BB-Cl (5 μM) or GSK199 (20 μM). Viral particle production is determined by plaque assay. Data are expressed as mean ± SEM from four independent experiments and analyzed by t test. ∗ p < 0.05, ∗∗ p < 0.01, and ∗∗∗ p < 0.001.

    Article Snippet: The following primary antibodies were sequentially applied to the slides: SARS-CoV-1/2 Spike Protein (clone 2B3E5, Cell Signaling), mouse anti-mouse β-tubulin IV (clone T7941, Merck), rat anti-mouse F4/80 (clone CI:A3-1, Bio-Rad Laboratories), rabbit anti-mouse PAD2 (polyclonal, Proteintech), rabbit anti-mouse PAD4 (polyclonal, Proteintech), rabbit anti-mouse Ly6g (clone EPR22909-135, Abcam), rat anti-mouse CD3 (clone CD3-12, Abcam), and rabbit anti-mouse cytokeratin 8 (clone EP1628Y, Abcam).

    Techniques: In Vitro, Plaque Assay, Concentration Assay, MTT Assay

    PAD4 inhibition blocks SARS-CoV-2 protein and genome synthesis (A) Relative viral RNA quantification in cell extracts from SARS-CoV-2-infected Huh7.5 (left) or Calu3 (right) cells (MOI 0.1) treated with GSK199 (20 μM), BB-Cl (5 μM), or DMSO, determined by comparative RT-qPCR. Values are expressed relative to vehicle-treated samples and normalized to the housekeeping gene PGK1. Data represent mean ± SEM from three independent experiments and are analyzed by t test. ∗ p < 0.05, ∗∗ p < 0.01, and ∗∗∗ p < 0.001. (B) Western blot analysis of viral protein expression in Huh7.5 and Calu3 cells, either mock-infected or infected with SARS-CoV-2 (MOI 0.1) and treated with GSK199 (20 μM), BB-Cl (5 μM), or DMSO. One representative blot from three independent experiments is shown. (C) Schematic representation of infection and treatment protocols used for the viral entry inhibition assay. (D) Viral entry assay on VERO-E6 (left) and TMPRSS2-rexpressing VERO-E6 (right) infected with VSV-Spike GFP (MOI 1). GFP-positive infected cells are microscopically counted, and the results are expressed as a percentage relative to vehicle-treated cells, and are analyzed by t test. Data represent mean ± SEM. (E) Absolute quantification of viral RNA released from SARS-CoV-2-infected Huh7.5 (left) or Calu3 (right) cells (MOI 0.1) treated with DMSO (vehicle) or GSK199 (20 μM), either using standard treatment (total) or added at 2 hpi (Post), determined by qRT-PCR. Supernatants are collected at 24 hpi for Calu3 and 48 hpi for Huh7.5. Data represent mean ± SEM from three independent experiments and are analyzed by t test. ∗ p < 0.05 and ∗∗ p < 0.01.

    Journal: iScience

    Article Title: Targeting peptidyl-arginine deiminase 4 suppresses SARS-CoV-2 replication and modulates the inflammatory response

    doi: 10.1016/j.isci.2026.115038

    Figure Lengend Snippet: PAD4 inhibition blocks SARS-CoV-2 protein and genome synthesis (A) Relative viral RNA quantification in cell extracts from SARS-CoV-2-infected Huh7.5 (left) or Calu3 (right) cells (MOI 0.1) treated with GSK199 (20 μM), BB-Cl (5 μM), or DMSO, determined by comparative RT-qPCR. Values are expressed relative to vehicle-treated samples and normalized to the housekeeping gene PGK1. Data represent mean ± SEM from three independent experiments and are analyzed by t test. ∗ p < 0.05, ∗∗ p < 0.01, and ∗∗∗ p < 0.001. (B) Western blot analysis of viral protein expression in Huh7.5 and Calu3 cells, either mock-infected or infected with SARS-CoV-2 (MOI 0.1) and treated with GSK199 (20 μM), BB-Cl (5 μM), or DMSO. One representative blot from three independent experiments is shown. (C) Schematic representation of infection and treatment protocols used for the viral entry inhibition assay. (D) Viral entry assay on VERO-E6 (left) and TMPRSS2-rexpressing VERO-E6 (right) infected with VSV-Spike GFP (MOI 1). GFP-positive infected cells are microscopically counted, and the results are expressed as a percentage relative to vehicle-treated cells, and are analyzed by t test. Data represent mean ± SEM. (E) Absolute quantification of viral RNA released from SARS-CoV-2-infected Huh7.5 (left) or Calu3 (right) cells (MOI 0.1) treated with DMSO (vehicle) or GSK199 (20 μM), either using standard treatment (total) or added at 2 hpi (Post), determined by qRT-PCR. Supernatants are collected at 24 hpi for Calu3 and 48 hpi for Huh7.5. Data represent mean ± SEM from three independent experiments and are analyzed by t test. ∗ p < 0.05 and ∗∗ p < 0.01.

    Article Snippet: The following primary antibodies were sequentially applied to the slides: SARS-CoV-1/2 Spike Protein (clone 2B3E5, Cell Signaling), mouse anti-mouse β-tubulin IV (clone T7941, Merck), rat anti-mouse F4/80 (clone CI:A3-1, Bio-Rad Laboratories), rabbit anti-mouse PAD2 (polyclonal, Proteintech), rabbit anti-mouse PAD4 (polyclonal, Proteintech), rabbit anti-mouse Ly6g (clone EPR22909-135, Abcam), rat anti-mouse CD3 (clone CD3-12, Abcam), and rabbit anti-mouse cytokeratin 8 (clone EP1628Y, Abcam).

    Techniques: Inhibition, Infection, Quantitative RT-PCR, Western Blot, Expressing, Quantitative Proteomics

    In vivo antiviral activity of PAD inhibitors against SARS-CoV-2 (A) Schematic representation of the treatment and infection protocols in K18-hACE2 transgenic mice. (B) Quantification of viral genome copies in mouse lungs—treated and infected as described in A— by ddPCR. Each dot represents an individual mouse sample ( n = 6). (C) Plaque assay to determine the number of infectious viral particles in mouse nasal swabs. (D) Graphical representation of cumulative scores from for immunohistochemical staining of mouse lungs using an anti-SARS-CoV-2 nucleocapsid antibody ( n = 4). Y axis represents the cumulative score calculated according to the criteria described in . (E) Graphical representation of cumulative scores from for H&E staining of mouse lungs ( n = 4), showing histopathological alterations classified by anatomical site and summarized as a percentage of tissue involvement. (F) Quantification of viral genome copies in mouse brains—treated and infected, as represented in A— by ddPCR. Each dot represents an individual mouse sample ( n = 6). (G) Distribution of brains from vehicle- or PAD-inhibitor-treated infected mice based on viral load (low: < below 10 3 ; high: > below 10 3 ). (H) Graphical representation of cumulative scores from for immunohistochemical staining of mouse brains using anti-SARS-CoV-2 nucleocapsid antibody ( n = 4). Y axis represents the cumulative score calculated according to the criteria described in . (I) Graphical representation of cumulative scores from for H&E staining of mouse brains ( n = 4), showing histopathological alterations classified by anatomical site and summarized as a percentage of tissue involvement. Data from all experiments, which represent mean ± SEM, are analyzed using an unpaired two-tailed t test. ∗ p < 0.05, ∗∗ p < 0.01, and ∗∗∗ p < 0.001.

    Journal: iScience

    Article Title: Targeting peptidyl-arginine deiminase 4 suppresses SARS-CoV-2 replication and modulates the inflammatory response

    doi: 10.1016/j.isci.2026.115038

    Figure Lengend Snippet: In vivo antiviral activity of PAD inhibitors against SARS-CoV-2 (A) Schematic representation of the treatment and infection protocols in K18-hACE2 transgenic mice. (B) Quantification of viral genome copies in mouse lungs—treated and infected as described in A— by ddPCR. Each dot represents an individual mouse sample ( n = 6). (C) Plaque assay to determine the number of infectious viral particles in mouse nasal swabs. (D) Graphical representation of cumulative scores from for immunohistochemical staining of mouse lungs using an anti-SARS-CoV-2 nucleocapsid antibody ( n = 4). Y axis represents the cumulative score calculated according to the criteria described in . (E) Graphical representation of cumulative scores from for H&E staining of mouse lungs ( n = 4), showing histopathological alterations classified by anatomical site and summarized as a percentage of tissue involvement. (F) Quantification of viral genome copies in mouse brains—treated and infected, as represented in A— by ddPCR. Each dot represents an individual mouse sample ( n = 6). (G) Distribution of brains from vehicle- or PAD-inhibitor-treated infected mice based on viral load (low: < below 10 3 ; high: > below 10 3 ). (H) Graphical representation of cumulative scores from for immunohistochemical staining of mouse brains using anti-SARS-CoV-2 nucleocapsid antibody ( n = 4). Y axis represents the cumulative score calculated according to the criteria described in . (I) Graphical representation of cumulative scores from for H&E staining of mouse brains ( n = 4), showing histopathological alterations classified by anatomical site and summarized as a percentage of tissue involvement. Data from all experiments, which represent mean ± SEM, are analyzed using an unpaired two-tailed t test. ∗ p < 0.05, ∗∗ p < 0.01, and ∗∗∗ p < 0.001.

    Article Snippet: The following primary antibodies were sequentially applied to the slides: SARS-CoV-1/2 Spike Protein (clone 2B3E5, Cell Signaling), mouse anti-mouse β-tubulin IV (clone T7941, Merck), rat anti-mouse F4/80 (clone CI:A3-1, Bio-Rad Laboratories), rabbit anti-mouse PAD2 (polyclonal, Proteintech), rabbit anti-mouse PAD4 (polyclonal, Proteintech), rabbit anti-mouse Ly6g (clone EPR22909-135, Abcam), rat anti-mouse CD3 (clone CD3-12, Abcam), and rabbit anti-mouse cytokeratin 8 (clone EP1628Y, Abcam).

    Techniques: In Vivo, Activity Assay, Infection, Transgenic Assay, Plaque Assay, Immunohistochemical staining, Staining, Two Tailed Test

    PAD inhibition modulates SARS-CoV-2-induced inflammation (A–D) Relative mRNA levels of the indicated genes are quantified by comparative RT-qPCR from total RNA of Calu-3 cells infected with the SARS-CoV-2 Delta strain (gray bars; MOI 0.1, 24 hpi) and treated with GSK199 (20 μM), BB-Cl (5 μM), or DMSO (vehicle). Data from three independent experiments were normalized to the housekeeping gene PGK1 and plotted as mean fold change ±SEM over mock-infected cells (white bars). (∗ p < 0.05, ∗∗ p < 0.01, and ∗∗∗ p < 0.001; one-way ANOVA followed by Bonferroni’s post-test). Red asterisks indicate comparisons with mock control, while black asterisks refer to comparisons among treatments. (E–J) Relative mRNA levels of the indicated cytokines are quantified by RT-qPCR from lung tissues of mice infected with SARS-CoV-2 Delta variant (10 5 PFU, i.n.) for 4 days and treated with BB-Cl (1 mg/kg), GSK199 (30 mg/kg), or DMSO (vehicle). Each dot represents one individual mouse. Data are normalized to the housekeeping gene β-actin and are presented relative to the mean ΔCt value of vehicle-treated mice. Statistical significance is assessed using an unpaired two-tailed t test (∗ p < 0.05, ∗∗ p < 0.01, and ∗∗∗ p < 0.001).

    Journal: iScience

    Article Title: Targeting peptidyl-arginine deiminase 4 suppresses SARS-CoV-2 replication and modulates the inflammatory response

    doi: 10.1016/j.isci.2026.115038

    Figure Lengend Snippet: PAD inhibition modulates SARS-CoV-2-induced inflammation (A–D) Relative mRNA levels of the indicated genes are quantified by comparative RT-qPCR from total RNA of Calu-3 cells infected with the SARS-CoV-2 Delta strain (gray bars; MOI 0.1, 24 hpi) and treated with GSK199 (20 μM), BB-Cl (5 μM), or DMSO (vehicle). Data from three independent experiments were normalized to the housekeeping gene PGK1 and plotted as mean fold change ±SEM over mock-infected cells (white bars). (∗ p < 0.05, ∗∗ p < 0.01, and ∗∗∗ p < 0.001; one-way ANOVA followed by Bonferroni’s post-test). Red asterisks indicate comparisons with mock control, while black asterisks refer to comparisons among treatments. (E–J) Relative mRNA levels of the indicated cytokines are quantified by RT-qPCR from lung tissues of mice infected with SARS-CoV-2 Delta variant (10 5 PFU, i.n.) for 4 days and treated with BB-Cl (1 mg/kg), GSK199 (30 mg/kg), or DMSO (vehicle). Each dot represents one individual mouse. Data are normalized to the housekeeping gene β-actin and are presented relative to the mean ΔCt value of vehicle-treated mice. Statistical significance is assessed using an unpaired two-tailed t test (∗ p < 0.05, ∗∗ p < 0.01, and ∗∗∗ p < 0.001).

    Article Snippet: The following primary antibodies were sequentially applied to the slides: SARS-CoV-1/2 Spike Protein (clone 2B3E5, Cell Signaling), mouse anti-mouse β-tubulin IV (clone T7941, Merck), rat anti-mouse F4/80 (clone CI:A3-1, Bio-Rad Laboratories), rabbit anti-mouse PAD2 (polyclonal, Proteintech), rabbit anti-mouse PAD4 (polyclonal, Proteintech), rabbit anti-mouse Ly6g (clone EPR22909-135, Abcam), rat anti-mouse CD3 (clone CD3-12, Abcam), and rabbit anti-mouse cytokeratin 8 (clone EP1628Y, Abcam).

    Techniques: Inhibition, Quantitative RT-PCR, Infection, Control, Variant Assay, Two Tailed Test

    PAD inhibition protects against SARS-CoV-2-induced pathological changes (A–C) Volcano plots show citrullinated proteins in pooled protein lysates from SARS-CoV-2- vs. mock-infected mouse lungs at 4 dpi. Lysates are incubated with biotin-PG to isolate citrullinated proteins on streptavidin agarose, followed by on-bead tryptic digestion and LC-MS/MS analysis. Each dot represents an identified citrullinated protein. The x axis shows the citrullination ratio between mock and infected samples (A) and between infected mice treated with vehicle (DMSO) or with PAD inhibitors BB-Cl-amidine (B) or GSK199 (C), while the y axis indicates statistical significance, both on a logarithmic scale ( n = 3). (D) Heatmap shows the fold changes of differentially citrullinated proteins identified in lung tissues across the indicated pairwise comparisons: uninfected vs. infected, and vehicle-treated infected vs. BB-Cl–treated or GSK199–treated infected mice. The color scale represents relative citrullination levels expressed as log 2 fold change.

    Journal: iScience

    Article Title: Targeting peptidyl-arginine deiminase 4 suppresses SARS-CoV-2 replication and modulates the inflammatory response

    doi: 10.1016/j.isci.2026.115038

    Figure Lengend Snippet: PAD inhibition protects against SARS-CoV-2-induced pathological changes (A–C) Volcano plots show citrullinated proteins in pooled protein lysates from SARS-CoV-2- vs. mock-infected mouse lungs at 4 dpi. Lysates are incubated with biotin-PG to isolate citrullinated proteins on streptavidin agarose, followed by on-bead tryptic digestion and LC-MS/MS analysis. Each dot represents an identified citrullinated protein. The x axis shows the citrullination ratio between mock and infected samples (A) and between infected mice treated with vehicle (DMSO) or with PAD inhibitors BB-Cl-amidine (B) or GSK199 (C), while the y axis indicates statistical significance, both on a logarithmic scale ( n = 3). (D) Heatmap shows the fold changes of differentially citrullinated proteins identified in lung tissues across the indicated pairwise comparisons: uninfected vs. infected, and vehicle-treated infected vs. BB-Cl–treated or GSK199–treated infected mice. The color scale represents relative citrullination levels expressed as log 2 fold change.

    Article Snippet: The following primary antibodies were sequentially applied to the slides: SARS-CoV-1/2 Spike Protein (clone 2B3E5, Cell Signaling), mouse anti-mouse β-tubulin IV (clone T7941, Merck), rat anti-mouse F4/80 (clone CI:A3-1, Bio-Rad Laboratories), rabbit anti-mouse PAD2 (polyclonal, Proteintech), rabbit anti-mouse PAD4 (polyclonal, Proteintech), rabbit anti-mouse Ly6g (clone EPR22909-135, Abcam), rat anti-mouse CD3 (clone CD3-12, Abcam), and rabbit anti-mouse cytokeratin 8 (clone EP1628Y, Abcam).

    Techniques: Inhibition, Infection, Incubation, Liquid Chromatography with Mass Spectroscopy