sars cov 2 2019 ncov nucleocapsid antibody rabbit pab (Sino Biological)


Name:
SARS CoV 2 2019 nCoV Nucleocapsid Antibody Rabbit PAb
Description:
Produced in rabbits immunized with purified recombinant SARS CoV 2 2019 nCoV Nucleocapsid Protein Catalog 40588 V08B YP 009724397 2 335Gly Ala Met1 Ala419 The specific IgG was purified by SARS CoV 2 2019 nCoV Nucleocapsid affinity chromatography
Catalog Number:
40588-t62
Product Aliases:
Anti-coronavirus NP Antibody, Anti-coronavirus Nucleocapsid Antibody, Anti-coronavirus Nucleoprotein Antibody, Anti-cov np Antibody, Anti-ncov NP Antibody, Anti-NCP-CoV Nucleocapsid Antibody, Anti-novel coronavirus NP Antibody, Anti-novel coronavirus Nucleocapsid Antibody, Anti-novel coronavirus Nucleoprotein Antibody, Anti-np Antibody, Anti-nucleocapsid Antibody, Anti-Nucleoprotein Antibody
Price:
None
Applications:
WB,ELISA
Host:
Rabbit
Immunogen:
Recombinant SARS-CoV-2 / 2019-nCoV Nucleocapsid Protein (Catalog#40588-V08B)
Category:
Primary Antibody
Antibody Type:
PAb
Isotype:
Rabbit IgG
Reactivity:
2019 nCoV
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Structured Review

Produced in rabbits immunized with purified recombinant SARS CoV 2 2019 nCoV Nucleocapsid Protein Catalog 40588 V08B YP 009724397 2 335Gly Ala Met1 Ala419 The specific IgG was purified by SARS CoV 2 2019 nCoV Nucleocapsid affinity chromatography
https://www.bioz.com/result/sars cov 2 2019 ncov nucleocapsid antibody rabbit pab/product/Sino Biological
Average 94 stars, based on 10 article reviews
Price from $9.99 to $1999.99
Images
1) Product Images from "ILRUN downregulates ACE2 expression and blocks infection of human cells by SARS-CoV-2"
Article Title: ILRUN downregulates ACE2 expression and blocks infection of human cells by SARS-CoV-2
Journal: bioRxiv
doi: 10.1101/2020.11.13.381343

Figure Legend Snippet: ILRUN supresses SARS-CoV-2 infection and down-regulates host genes essential for SARS-CoV-2 entry. (A) Transcription profile of SARS-CoV-2 in Caco-2 cells transfected with 40 nM siNEG or siILRUN for 72 h at 6 h and 24 h post infection. (B) SARS-CoV-2 titres of supernatants from Caco-2 cells infected with SARS-CoV-2 (24 h, MOI 0.3) post-transfection with siRNAs (40 nM, 72 h) *p
Techniques Used: Infection, Transfection

Figure Legend Snippet: Validation of ILRUN function and SARS-CoV-2 infection in Caco-2 cells. (A) ILRUN mRNA levels (2 −ΔΔCt relative to GAPDH ) in Caco-2 cells transfected with siRNAs (40 nM, 72 h) targeting ILRUN or a nontargeting control (siNEG). **p
Techniques Used: Infection, Transfection
2) Product Images from "Functional immune mapping with deep-learning enabled phenomics applied to immunomodulatory and COVID-19 drug discovery"
Article Title: Functional immune mapping with deep-learning enabled phenomics applied to immunomodulatory and COVID-19 drug discovery
Journal: bioRxiv
doi: 10.1101/2020.08.02.233064

Figure Legend Snippet: Repurposed library screening for COVID-19 using phenomics A . Syk, c-Met and PI3K inhibitors rescue the severe COVID-19 specific cytokine storm high-dimensional phenoprint (perturbed state) to the healthy phenoprint (target state). B . Example images of target and perturbed cell populations for the cytokine storm and SARS-CoV2 viral models. C . Infection of HRCE yielded a phenoprint against the mock-infected target population with an assay z-factor of 0.43 for the separation in on-perturbation score for the mock and infected populations. D-F . Projections of compound response in the context of the perturbation vector generated in SARS-CoV-2-infected HRCE, Vero, and Calu3 cells. Off-perturbation values clipped at 50 for visualization. G . Compound impact on endothelial barrier function as quantified by ECIS assay. Values are normalized from 0 (cytokine storm cocktail-treated wells) to 100 (mock-treated wells). Data was averaged over a 12-minute window at hour 12 of ECIS measurement to visualize concentration response curves for the indicated compounds. H . Infection rate as determined by SARS-CoV-2 nucleocapsid antibody staining of infected HRCEs treated with the denoted compounds. I . Plot of efficacious molecules by hit-scores in SARS-CoV-2 HRCE assay vs cytokine storm assay. Orange circles denote molecules registered in interventional COVID-19 clinical trials at the time of submission. Dotted lines presented as a visual guide depicting a hit score of 0.6.
Techniques Used: Library Screening, Infection, Plasmid Preparation, Generated, Electric Cell-substrate Impedance Sensing, Concentration Assay, Staining

Figure Legend Snippet: SARS-CoV-2 infection model A . Quantification of active SARS-CoV-2 production over time in the indicated cell types using TCID50 measurement on Vero cells (n=2). B . Representative images of HRCE, Calu3 and Vero cells immunostained with SARS-CoV-2 nucleocapsid protein (pink) and modified cell paint dyes C . Infection rates of each tested cell type as analyzed by nucleocapsid immunostaining. Of note, HRCE donors displayed significant variation in infectability and only a minority of donors exhibited infection rates high enough for screening. Antibody stains were performed after the principal analysis concluded, and are therefore not represented in the primary dataset used for phenoprint evaluation and compound screening. D . Infection of HRCE yielded a phenoprint against the mock-infected target population with an assay z-factor of 0.43 and was selected for further investigation. Vero and Calu3 cells also demonstrated screenable phenoprints. E . Quantification of percentage of cells infected using nucleocapsid protein immunostaining in Calu3 cells at 96 hours post infection for key compounds F . Consistency of hit scores for selected compounds across HRCE donors and between cell types. G . Projections of compound response of JAK inhibitor and control compounds onto the perturbation vector generated in SARS-CoV-2-infected HRCE. H . Quantification of percent of cells infected using nucleocapsid protein immunostaining in HRCE cells at 96 hours post infection for JAK inhibitors
Techniques Used: Infection, Modification, Immunostaining, Plasmid Preparation, Generated
3) Product Images from "Functional immune mapping with deep-learning enabled phenomics applied to immunomodulatory and COVID-19 drug discovery"
Article Title: Functional immune mapping with deep-learning enabled phenomics applied to immunomodulatory and COVID-19 drug discovery
Journal: bioRxiv
doi: 10.1101/2020.08.02.233064

Figure Legend Snippet: Repurposed library screening for COVID-19 using phenomics A . Syk, c-Met and PI3K inhibitors rescue the severe COVID-19 specific cytokine storm high-dimensional phenoprint (perturbed state) to the healthy phenoprint (target state). B . Example images of target and perturbed cell populations for the cytokine storm and SARS-CoV2 viral models. C . Infection of HRCE yielded a phenoprint against the mock-infected target population with an assay z-factor of 0.43 for the separation in on-perturbation score for the mock and infected populations. D-F . Projections of compound response in the context of the perturbation vector generated in SARS-CoV-2-infected HRCE, Vero, and Calu3 cells. Off-perturbation values clipped at 50 for visualization. G . Compound impact on endothelial barrier function as quantified by ECIS assay. Values are normalized from 0 (cytokine storm cocktail-treated wells) to 100 (mock-treated wells). Data was averaged over a 12-minute window at hour 12 of ECIS measurement to visualize concentration response curves for the indicated compounds. H . Infection rate as determined by SARS-CoV-2 nucleocapsid antibody staining of infected HRCEs treated with the denoted compounds. I . Plot of efficacious molecules by hit-scores in SARS-CoV-2 HRCE assay vs cytokine storm assay. Orange circles denote molecules registered in interventional COVID-19 clinical trials at the time of submission. Dotted lines presented as a visual guide depicting a hit score of 0.6.
Techniques Used: Library Screening, Infection, Plasmid Preparation, Generated, Electric Cell-substrate Impedance Sensing, Concentration Assay, Staining

Figure Legend Snippet: SARS-CoV-2 infection model A . Quantification of active SARS-CoV-2 production over time in the indicated cell types using TCID50 measurement on Vero cells (n=2). B . Representative images of HRCE, Calu3 and Vero cells immunostained with SARS-CoV-2 nucleocapsid protein (pink) and modified cell paint dyes C . Infection rates of each tested cell type as analyzed by nucleocapsid immunostaining. Of note, HRCE donors displayed significant variation in infectability and only a minority of donors exhibited infection rates high enough for screening. Antibody stains were performed after the principal analysis concluded, and are therefore not represented in the primary dataset used for phenoprint evaluation and compound screening. D . Infection of HRCE yielded a phenoprint against the mock-infected target population with an assay z-factor of 0.43 and was selected for further investigation. Vero and Calu3 cells also demonstrated screenable phenoprints. E . Quantification of percentage of cells infected using nucleocapsid protein immunostaining in Calu3 cells at 96 hours post infection for key compounds F . Consistency of hit scores for selected compounds across HRCE donors and between cell types. G . Projections of compound response of JAK inhibitor and control compounds onto the perturbation vector generated in SARS-CoV-2-infected HRCE. H . Quantification of percent of cells infected using nucleocapsid protein immunostaining in HRCE cells at 96 hours post infection for JAK inhibitors
Techniques Used: Infection, Modification, Immunostaining, Plasmid Preparation, Generated
4) Product Images from "An effective, safe and cost-effective cell-based chimeric vaccine against SARS-CoV2"
Article Title: An effective, safe and cost-effective cell-based chimeric vaccine against SARS-CoV2
Journal: bioRxiv
doi: 10.1101/2020.08.19.258244

Figure Legend Snippet: The immunogenicity, efficacy and safety of C-Vac for SARS-CoV-2 infection. (A) Flow cytometry-based detection of MHC(HLA-A2)-peptide complex binding affinity in T2 cells. (B) The expression of antigens in 293T-based C-Vac is confirmed by Western Blot. N, 293T cells transfected with the plasmid expressing a full length N gene. (C D) Pseudovirus neutralization titers of hamster serum at day 7 after the first immunization and day 21 (boosted at day 14) after vaccination with 293T-based C-Vac, MiT C-Vac: Mitomycin C-treated C-Vac, Lys C-Vac: Lysed C-Vac. (E) Histological characteristics of hamster lung at day 7 after the first vaccination. Original magnification 200× (F) Allograft volume of transformed fibroblasts expressing RBD-truncated N protein in the Syrian hamsters immunized with different regime. 5×10 6 BHK21 cells expressing C-Vac antigen (RBD-Ntap) were subcutaneously injected into immunized hamsters for challenge at day 45 after boost, and the volume of allografts were measured at 14 days after inoculation of the BHK21 cells expressing RBD-Ntap into the immunized hamsters.
Techniques Used: Infection, Flow Cytometry, Binding Assay, Expressing, Western Blot, Transfection, Plasmid Preparation, Neutralization, Transformation Assay, Injection

Figure Legend Snippet: The RBD domain of Spike is crucial for the SARS-CoV2 Vaccine. (A) The functional domain of SARS-CoV-2 spike protein. (B) Potential B cell antigen of RBD domain from SARS-CoV2 is predicted by Discotope software based on their 3D structure. (C) Potential linear B cell epitopes of SARS-CoV-2 full S protein are analysed with the IEDB database. (D) The location of potential antigens in RBD domain (SARS-CoV-2:red, SARS-CoV: Purple) and interaction model between RBD and ACE2 receptor (interface is marked yellow) are marked with Discovery Studio. (E) The expression of Spike and nucleocapsid with wild-type sequence in 293T cells are detected by Western Blot assay. (F) The expression of Spike and its derivatives with codon optimization (opt).
Techniques Used: Functional Assay, Software, Expressing, Sequencing, Western Blot

Figure Legend Snippet: Construction of chimeric vaccine for SARS-CoV-2. (A) Potential B-cell epitopes of N protein is predicted by IEDB database. (B) Potential MHCI-binding peptides of N. (C) Functional domain of SARS-CoV N protein (Upper) and its antibody epitope map reported in previous study. (D) The skeleton of Chimeric Vaccine for SARS-CoV-2, RBD: spike RBD domain (306-541 aa), Ntap: T-cell-associated peptide of N (211-339 aa). (E) Characterization of SARS-CoV-2-derived protein and C-Vac antigen by SARS-CoV-2 antisera and commercial antibodies against SARS-CoV2 spike RBD or Nucleocapsid.
Techniques Used: Binding Assay, Functional Assay, Derivative Assay
5) Product Images from "Heterogeneous antibodies against SARS-CoV-2 spike receptor binding domain and nucleocapsid with implications for COVID-19 immunity"
Article Title: Heterogeneous antibodies against SARS-CoV-2 spike receptor binding domain and nucleocapsid with implications for COVID-19 immunity
Journal: JCI Insight
doi: 10.1172/jci.insight.142386

Figure Legend Snippet: Comparison of seroconversion in patients with COVID-19 and healthy individuals. ( A ) ELISA with S-RBD protein coating and 1:100 dilution of repeated serum samples of patients with SARS-CoV-2 and healthy individuals. Absorbance normalized to the respective no antigen control for each sample at 450 nm reported. SARS-CoV-2 (blue), n = 88 (from 21 patients); HS 2017–2019 (white), n = 104; HS 2020 (white), n = 308. Arrows list consecutive serum samples evaluated for each case. Inset graphs depict the data separated based on healthy serum collected from 2017 to 2019 (left inset) and 2020 (right inset). ( B ) ELISA with N-protein coating and 1:100 dilution of the first and last serum samples of patients with SARS-CoV-2 and healthy individuals. Absorbance normalized to the respective no antigen control for each sample at 450 nm reported. SARS-CoV-2 (blue), n = 37 (from 21 patients); HS 2017–2019 (white), n = 103; HS 2020 (white), n = 308. Arrows list consecutive serum samples evaluated for each case. Inset graphs depict the data separated based on healthy serum collected from 2017 to 2019 (top inset) and 2020 (bottom inset). ( C ) Pie charts depicting percentage of samples positive for indicated antigens. SARS-CoV-2, n = 21; HS 2017–2019, n = 103; HS 2020, n = 308; non–COVID-19 samples (NCSs), n = 45; HIV, n = 7; all, n = 484.
Techniques Used: Enzyme-linked Immunosorbent Assay

Figure Legend Snippet: Detection of serum binding antibodies against SARS-CoV-2 proteins in patients with PCR-confirmed COVID-19 and healthy samples. ( A ) Timeline of COVID-19 diagnosis/ICU admittance, serum sample collection, and convalescent plasma (CP) administration. Time 0 is defined as day of COVID-19 diagnosis (PCR positive for SARS-CoV-2) and ICU admittance. Blood collections are denoted in gray and CP administration is denoted in pink. Patients were stratified based on current status (recovered, hospitalized, or deceased). Patient 29 from our cohort had symptoms but was PCR negative for SARS-CoV-2; this sample was not included in figures since there was no proof of disease. ( B ) Schematic of SARS-CoV-2 viral structure (top panel) and antigens assayed (bottom panel). S-protein, light orange; envelope protein, yellow; membrane glycoprotein, dark orange; RNA, blue; N-protein, green. Absorbance normalized to the respective no antigen control for each sample at 450 nm plotted for S-RBD (left panel), and N-protein (right panel), antigen coating with the most recent (or only) SARS-CoV-2 samples not treated with CP ( n = 21) and healthy samples collected in 2017–2019 (HS 2017–2019, n = 104 for S-RBD, n = 103 for N-protein) and 2020 (HS 2020, n = 308). Data are presented with each dot representing the mean normalized absorbance for a given serum sample; the red bar depicts the median ± interquartile range of all samples. HS, healthy sample; NC (line), negative control cutoff (see Methods). Kruskal-Wallis with Dunn’s multiple-comparisons test performed. **** P
Techniques Used: Binding Assay, Polymerase Chain Reaction, Negative Control

Figure Legend Snippet: Pseudotyped SARS-CoV-2 virion neutralization activity of serum binding antibodies against S-RBD and N-protein. ( A ) Luminescence normalized to FBS+Virus control obtained from pseudovirus neutralization assay at 1:20 serum dilution. ( B ) Matched serological results for S-RBD at 1:100 serum dilution (top 2 panels) and 1:20 serum dilution (bottom 2 panels). Absorbance normalized to the respective no antigen control for each sample at 450 nm reported. Case numbers are color-coded: green: recovered, red: deceased, blue: hospitalized. ( C ) Matched serological results for N-protein at 1:100 serum dilution and 1:20 serum dilution. Absorbance normalized to the respective no antigen control for each sample at 450 nm reported. Case numbers are color-coded: green: recovered, red: deceased, blue: hospitalized. Data ( A – C ) are reported as mean ± standard deviation (SD) of 3 technical replicates for each sample. ( D ) Heatmap depicting positive and negative categorization of the listed serum cases for each viral protein tested in serological and neutr3alization assays. Low titer positive as defined by detecting of binding antibodies shown in Figure 2, C and D , 1:20 titer.
Techniques Used: Neutralization, Activity Assay, Binding Assay, Standard Deviation
6) Product Images from "SARS-CoV-2 and Three Related Coronaviruses Utilize Multiple ACE2 Orthologs and Are Potently Blocked by an Improved ACE2-Ig"
Article Title: SARS-CoV-2 and Three Related Coronaviruses Utilize Multiple ACE2 Orthologs and Are Potently Blocked by an Improved ACE2-Ig
Journal: Journal of Virology
doi: 10.1128/JVI.01283-20

Figure Legend Snippet: A wide range of ACE2 orthologs support binding to RBD proteins of SARS-CoV-2 and three related coronaviruses. (A) 293T cells were transfected with adjusted amounts of the indicated ACE2-ortholog plasmids to have similar expression levels of the ACE2 ortholog proteins. Cells were then stained with an RBD-mouse IgG2 Fc fusion protein of SARS-CoV-2 WHU01, Pangolin-CoV-2020, Bat-CoV RaTG13, or SARS-CoV BJ01, followed by staining with an Alexa 488-goat anti-mouse IgG secondary antibody. RBD-ACE2 binding was detected using flow cytometry. (B) Percentages of cells positive for RBD binding in panel A are presented as a heatmap according to the indicated color code. (C) Expression levels of the indicated ACE2 orthologs were detected using Western blotting. The data shown are representative of two independent experiments performed by two different people with similar results.
Techniques Used: Binding Assay, Transfection, Expressing, Staining, Flow Cytometry, Western Blot

Figure Legend Snippet: A wide range of ACE2 orthologs support cell entry of SARS-CoV-2 and three related coronaviruses. (A to F) 293T cells in 96-well plates were transfected with adjusted amounts of the indicated ACE2-ortholog plasmids to have similar expression levels of the ACE2 ortholog proteins. Cells were then infected with retrovirus-based luciferase reporter pseudoviral particles (pp) enveloped with the indicated spike proteins. ACE2 ortholog-mediated viral entry was measured by luciferase reporter expression at 48 h (A to D and F) or 60 h (E) postinfection. (G) The relative infection (%) values for each ACE2 ortholog-mediated viral entry shown in panels A to F were independently calculated against the highest expression values of the same pseudotype panel and are presented as a heatmap according to the indicated color code. (H) 293T cells expressing ACE2 orthologs of the indicated species were infected with SARS-CoV-2 live virus at 800 TCID 50 . Cells were then fixed and stained with rabbit anti-SARS-CoV-2 nucleocapsid (NP) polyclonal antibody for fluorescence microscopy at 24 h postinfection. Red indicates SARS-CoV-2 NP, and blue indicates cell nuclei. Scale bars, 50 μm. The data shown are representative of two or three experiments independently performed by two different people with similar results, and data points in panels A to F represent the means ± the SD of four biological replicates.
Techniques Used: Transfection, Expressing, Infection, Luciferase, Staining, Fluorescence, Microscopy

Figure Legend Snippet: The 740-D30E variant of ACE2-Ig broadly neutralizes entry of SARS-CoV-2, SARS-CoV, Pangolin-CoV-2020 and Bat-CoV RaTG13. (A to D) Human ACE2-expressing 293T were infected with the indicated pseudotypes in the presence of an Fc fusion protein, F10-scFv (gray), ACE2 740-wt (blue), or ACE2 740-D30E (red). Viral entry was measured by luciferase reporter expression at 48 h (A, B, and D) or 60 h (C) postinfection, and the percent infection (Infection%) values were calculated. Note that the D30E mutation on the ACE2-Ig protein improved the protein’s neutralization activity against SARS-CoV-2 (A) and RaTG13 (C) but not Pangolin-CoV-2020 (B) or SARS-CoV (D). The dashed line in panels C and D indicates the background luciferase signals detected in the pseudovirus-infected parental 293T cells. (E) Human ACE2 residue D30 forms a salt bridge with the SARS-CoV-2 RBD residue K417 (PDB accession no. 6M0J ). SARS-CoV-2 and RaTG13 have a K417 residue at their spike proteins, while Pangolin-CoV has an R417 residue and SARS-CoV has a V417 residue at their spike proteins, respectively. Thus, a stabilized salt bridge interaction between E30 of the ACE2-Ig protein and K417 of the virus spike protein is likely responsible for the D30E mutation-mediated neutralization enhancement. The data shown are representative of two or three experiments independently performed by two different people with similar results, and data points in panels A to D represent the means ± the SD of three or four biological replicates.
Techniques Used: Variant Assay, Expressing, Infection, Luciferase, Mutagenesis, Neutralization, Activity Assay

Figure Legend Snippet: Recombinant RBD-Ig and ACE2-Ig variants efficiently block SARS-CoV-2 entry. (A) Diagrams of RBD-Ig and ACE2-Ig fusion proteins used in the following studies. (B and C) ACE2-expressing 293T cells were infected with SARS-CoV-2 spike-pseudotyped retrovirus (pp) in the presence of purified recombinant RBD-Ig (B) and ACE2-Ig (C) fusion proteins at the indicated concentrations. An Fc fusion protein of an anti-influenza HA antibody, F10-scFv, was used as a control protein here. Viral entry was measured by the luciferase reporter at 48 h postinfection. Luminescence values observed at each concentration were divided by the values observed at concentration zero to calculate the percent infection (Infection%) values. Note that all the 740-version variants showed significantly better potency than the 615-version variants (two-tailed two-sample t test, P
Techniques Used: Recombinant, Blocking Assay, Expressing, Infection, Purification, Luciferase, Concentration Assay, Two Tailed Test

Figure Legend Snippet: SARS-CoV-2 and ACE2 contact residues are conserved among four SARS-like viruses and 16 ACE2 orthologs, respectively. (A) Interactions between the SARS-CoV-2 receptor binding domain (RBD, red) and ACE2 (blue) involve a large number of contact residues (PDB accession no. 6M0J ). RBD residues
Techniques Used: Binding Assay

Figure Legend Snippet: A further improved ACE2-Ig variant with an antibody-like configuration potently neutralizes SARS-CoV-2 live virus. (A) Diagrams of ACE2-Ig variants characterized in the following studies. CH1, IgG heavy-chain constant region 1; CL, human antibody kappa light-chain constant region. (B and C) Human ACE2-expressing 293T (B) or HeLa (C) cells were infected with SARS-CoV-2 pseudotype in the presence of the indicated human IgG1 Fc fusion proteins at the indicated concentrations. An anti-HIV antibody b12 was used as a human IgG1 control. Viral entry was measured by luciferase reporter expression at 48 h postinfection, and the percent infection (Infection%) values were calculated. Estimated IC 50 and IC 90 values for each protein are directly derived from the curves and are shown to the right of the figures. (D) Human ACE2-expressing HeLa cells were infected with SARS-CoV-2 live virus at 800 TCID 50 in the presence of the b12 control protein, ACE2-Ig-v1, or ACE2-Ig-v3 at the indicated concentrations. Cells were then fixed and stained with rabbit anti-SARS-CoV-2 NP polyclonal antibody for fluorescence microscopy at 24 h postinfection. Red indicates SARS-CoV-2 NP and blue indicates cell nuclei. Scale bars, 200 μm. Note that ACE2-Ig-v3 at 0.8 μg/ml (1.85 nM) completely abolished viral NP signal. The data shown are representative of two or three experiments independently performed by two different people with similar results, and data points in panels B and C represent the means ± the SD of three biological replicates.
Techniques Used: Variant Assay, Expressing, Infection, Luciferase, Derivative Assay, Staining, Fluorescence, Microscopy
7) Product Images from "An effective, safe and cost-effective cell-based chimeric vaccine against SARS-CoV2"
Article Title: An effective, safe and cost-effective cell-based chimeric vaccine against SARS-CoV2
Journal: bioRxiv
doi: 10.1101/2020.08.19.258244

Figure Legend Snippet: The immunogenicity, efficacy and safety of C-Vac for SARS-CoV-2 infection. (A) Flow cytometry-based detection of MHC(HLA-A2)-peptide complex binding affinity in T2 cells. (B) The expression of antigens in 293T-based C-Vac is confirmed by Western Blot. N, 293T cells transfected with the plasmid expressing a full length N gene. (C D) Pseudovirus neutralization titers of hamster serum at day 7 after the first immunization and day 21 (boosted at day 14) after vaccination with 293T-based C-Vac, MiT C-Vac: Mitomycin C-treated C-Vac, Lys C-Vac: Lysed C-Vac. (E) Histological characteristics of hamster lung at day 7 after the first vaccination. Original magnification 200× (F) Allograft volume of transformed fibroblasts expressing RBD-truncated N protein in the Syrian hamsters immunized with different regime. 5×10 6 BHK21 cells expressing C-Vac antigen (RBD-Ntap) were subcutaneously injected into immunized hamsters for challenge at day 45 after boost, and the volume of allografts were measured at 14 days after inoculation of the BHK21 cells expressing RBD-Ntap into the immunized hamsters.
Techniques Used: Infection, Flow Cytometry, Binding Assay, Expressing, Western Blot, Transfection, Plasmid Preparation, Neutralization, Transformation Assay, Injection

Figure Legend Snippet: The RBD domain of Spike is crucial for the SARS-CoV2 Vaccine. (A) The functional domain of SARS-CoV-2 spike protein. (B) Potential B cell antigen of RBD domain from SARS-CoV2 is predicted by Discotope software based on their 3D structure. (C) Potential linear B cell epitopes of SARS-CoV-2 full S protein are analysed with the IEDB database. (D) The location of potential antigens in RBD domain (SARS-CoV-2:red, SARS-CoV: Purple) and interaction model between RBD and ACE2 receptor (interface is marked yellow) are marked with Discovery Studio. (E) The expression of Spike and nucleocapsid with wild-type sequence in 293T cells are detected by Western Blot assay. (F) The expression of Spike and its derivatives with codon optimization (opt).
Techniques Used: Functional Assay, Software, Expressing, Sequencing, Western Blot

Figure Legend Snippet: Construction of chimeric vaccine for SARS-CoV-2. (A) Potential B-cell epitopes of N protein is predicted by IEDB database. (B) Potential MHCI-binding peptides of N. (C) Functional domain of SARS-CoV N protein (Upper) and its antibody epitope map reported in previous study. (D) The skeleton of Chimeric Vaccine for SARS-CoV-2, RBD: spike RBD domain (306-541 aa), Ntap: T-cell-associated peptide of N (211-339 aa). (E) Characterization of SARS-CoV-2-derived protein and C-Vac antigen by SARS-CoV-2 antisera and commercial antibodies against SARS-CoV2 spike RBD or Nucleocapsid.
Techniques Used: Binding Assay, Functional Assay, Derivative Assay
8) Product Images from "Heterogeneous antibodies against SARS-CoV-2 spike receptor binding domain and nucleocapsid with implications for COVID-19 immunity"
Article Title: Heterogeneous antibodies against SARS-CoV-2 spike receptor binding domain and nucleocapsid with implications for COVID-19 immunity
Journal: JCI Insight
doi: 10.1172/jci.insight.142386

Figure Legend Snippet: Comparison of seroconversion in patients with COVID-19 and healthy individuals. ( A ) ELISA with S-RBD protein coating and 1:100 dilution of repeated serum samples of patients with SARS-CoV-2 and healthy individuals. Absorbance normalized to the respective no antigen control for each sample at 450 nm reported. SARS-CoV-2 (blue), n = 88 (from 21 patients); HS 2017–2019 (white), n = 104; HS 2020 (white), n = 308. Arrows list consecutive serum samples evaluated for each case. Inset graphs depict the data separated based on healthy serum collected from 2017 to 2019 (left inset) and 2020 (right inset). ( B ) ELISA with N-protein coating and 1:100 dilution of the first and last serum samples of patients with SARS-CoV-2 and healthy individuals. Absorbance normalized to the respective no antigen control for each sample at 450 nm reported. SARS-CoV-2 (blue), n = 37 (from 21 patients); HS 2017–2019 (white), n = 103; HS 2020 (white), n = 308. Arrows list consecutive serum samples evaluated for each case. Inset graphs depict the data separated based on healthy serum collected from 2017 to 2019 (top inset) and 2020 (bottom inset). ( C ) Pie charts depicting percentage of samples positive for indicated antigens. SARS-CoV-2, n = 21; HS 2017–2019, n = 103; HS 2020, n = 308; non–COVID-19 samples (NCSs), n = 45; HIV, n = 7; all, n = 484.
Techniques Used: Enzyme-linked Immunosorbent Assay

Figure Legend Snippet: Detection of serum binding antibodies against SARS-CoV-2 proteins in patients with PCR-confirmed COVID-19 and healthy samples. ( A ) Timeline of COVID-19 diagnosis/ICU admittance, serum sample collection, and convalescent plasma (CP) administration. Time 0 is defined as day of COVID-19 diagnosis (PCR positive for SARS-CoV-2) and ICU admittance. Blood collections are denoted in gray and CP administration is denoted in pink. Patients were stratified based on current status (recovered, hospitalized, or deceased). Patient 29 from our cohort had symptoms but was PCR negative for SARS-CoV-2; this sample was not included in figures since there was no proof of disease. ( B ) Schematic of SARS-CoV-2 viral structure (top panel) and antigens assayed (bottom panel). S-protein, light orange; envelope protein, yellow; membrane glycoprotein, dark orange; RNA, blue; N-protein, green. Absorbance normalized to the respective no antigen control for each sample at 450 nm plotted for S-RBD (left panel), and N-protein (right panel), antigen coating with the most recent (or only) SARS-CoV-2 samples not treated with CP ( n = 21) and healthy samples collected in 2017–2019 (HS 2017–2019, n = 104 for S-RBD, n = 103 for N-protein) and 2020 (HS 2020, n = 308). Data are presented with each dot representing the mean normalized absorbance for a given serum sample; the red bar depicts the median ± interquartile range of all samples. HS, healthy sample; NC (line), negative control cutoff (see Methods). Kruskal-Wallis with Dunn’s multiple-comparisons test performed. **** P
Techniques Used: Binding Assay, Polymerase Chain Reaction, Negative Control

Figure Legend Snippet: Pseudotyped SARS-CoV-2 virion neutralization activity of serum binding antibodies against S-RBD and N-protein. ( A ) Luminescence normalized to FBS+Virus control obtained from pseudovirus neutralization assay at 1:20 serum dilution. ( B ) Matched serological results for S-RBD at 1:100 serum dilution (top 2 panels) and 1:20 serum dilution (bottom 2 panels). Absorbance normalized to the respective no antigen control for each sample at 450 nm reported. Case numbers are color-coded: green: recovered, red: deceased, blue: hospitalized. ( C ) Matched serological results for N-protein at 1:100 serum dilution and 1:20 serum dilution. Absorbance normalized to the respective no antigen control for each sample at 450 nm reported. Case numbers are color-coded: green: recovered, red: deceased, blue: hospitalized. Data ( A – C ) are reported as mean ± standard deviation (SD) of 3 technical replicates for each sample. ( D ) Heatmap depicting positive and negative categorization of the listed serum cases for each viral protein tested in serological and neutr3alization assays. Low titer positive as defined by detecting of binding antibodies shown in Figure 2, C and D , 1:20 titer.
Techniques Used: Neutralization, Activity Assay, Binding Assay, Standard Deviation
9) Product Images from "Potential therapeutic effects of dipyridamole in the severely ill patients with COVID-19"
Article Title: Potential therapeutic effects of dipyridamole in the severely ill patients with COVID-19
Journal: Acta Pharmaceutica Sinica. B
doi: 10.1016/j.apsb.2020.04.008

Figure Legend Snippet: Suppressive effects of dipyridamole (DIP) and chloroquine on SARS-CoV-2 replication in vitro . (A) Chemical structure of DIP. (B) Enzyme activity of Mpro in the presence of ascending concentrations of DIP. (C) Dose-dependent suppression of SARS-CoV-2 replication by DIP and chloroquine in vitro . Virus titers were measured by Foci forming assay, inhibition rates were performed by indirect immunoinfluscent assay, and calculated inhibition rates of different dosages of DIP or chloroquine were compared with virus control. P values were calculated by ANOVA.
Techniques Used: In Vitro, Activity Assay, Inhibition
10) Product Images from "Functional immune mapping with deep-learning enabled phenomics applied to immunomodulatory and COVID-19 drug discovery"
Article Title: Functional immune mapping with deep-learning enabled phenomics applied to immunomodulatory and COVID-19 drug discovery
Journal: bioRxiv
doi: 10.1101/2020.08.02.233064

Figure Legend Snippet: Repurposed library screening for COVID-19 using phenomics A . Syk, c-Met and PI3K inhibitors rescue the severe COVID-19 specific cytokine storm high-dimensional phenoprint (perturbed state) to the healthy phenoprint (target state). B . Example images of target and perturbed cell populations for the cytokine storm and SARS-CoV2 viral models. C . Infection of HRCE yielded a phenoprint against the mock-infected target population with an assay z-factor of 0.43 for the separation in on-perturbation score for the mock and infected populations. D-F . Projections of compound response in the context of the perturbation vector generated in SARS-CoV-2-infected HRCE, Vero, and Calu3 cells. Off-perturbation values clipped at 50 for visualization. G . Compound impact on endothelial barrier function as quantified by ECIS assay. Values are normalized from 0 (cytokine storm cocktail-treated wells) to 100 (mock-treated wells). Data was averaged over a 12-minute window at hour 12 of ECIS measurement to visualize concentration response curves for the indicated compounds. H . Infection rate as determined by SARS-CoV-2 nucleocapsid antibody staining of infected HRCEs treated with the denoted compounds. I . Plot of efficacious molecules by hit-scores in SARS-CoV-2 HRCE assay vs cytokine storm assay. Orange circles denote molecules registered in interventional COVID-19 clinical trials at the time of submission. Dotted lines presented as a visual guide depicting a hit score of 0.6.
Techniques Used: Library Screening, Infection, Plasmid Preparation, Generated, Electric Cell-substrate Impedance Sensing, Concentration Assay, Staining

Figure Legend Snippet: SARS-CoV-2 infection model A . Quantification of active SARS-CoV-2 production over time in the indicated cell types using TCID50 measurement on Vero cells (n=2). B . Representative images of HRCE, Calu3 and Vero cells immunostained with SARS-CoV-2 nucleocapsid protein (pink) and modified cell paint dyes C . Infection rates of each tested cell type as analyzed by nucleocapsid immunostaining. Of note, HRCE donors displayed significant variation in infectability and only a minority of donors exhibited infection rates high enough for screening. Antibody stains were performed after the principal analysis concluded, and are therefore not represented in the primary dataset used for phenoprint evaluation and compound screening. D . Infection of HRCE yielded a phenoprint against the mock-infected target population with an assay z-factor of 0.43 and was selected for further investigation. Vero and Calu3 cells also demonstrated screenable phenoprints. E . Quantification of percentage of cells infected using nucleocapsid protein immunostaining in Calu3 cells at 96 hours post infection for key compounds F . Consistency of hit scores for selected compounds across HRCE donors and between cell types. G . Projections of compound response of JAK inhibitor and control compounds onto the perturbation vector generated in SARS-CoV-2-infected HRCE. H . Quantification of percent of cells infected using nucleocapsid protein immunostaining in HRCE cells at 96 hours post infection for JAK inhibitors
Techniques Used: Infection, Modification, Immunostaining, Plasmid Preparation, Generated
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