mers  (Sino Biological)


Bioz Verified Symbol Sino Biological is a verified supplier
Bioz Manufacturer Symbol Sino Biological manufactures this product  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 86
    Name:
    MERS CoV Spike Protein
    Description:
    A DNA sequence encoding the extracellular domain of spike protein MERS CoV AFS88936 1 Met1 Trp1297 was fused with a polyhistidine tag at the C terminus
    Catalog Number:
    40069-V08B
    Price:
    None
    Category:
    recombinant protein
    Product Aliases:
    coronavirus s1 Protein MERS-CoV, coronavirus s2 Protein MERS-CoV, coronavirus spike Protein MERS-CoV, cov spike Protein MERS-CoV, ncov RBD Protein MERS-CoV, ncov s1 Protein MERS-CoV, ncov s2 Protein MERS-CoV, ncov spike Protein MERS-CoV, RBD Protein MERS-CoV, S Protein MERS-CoV, s1 Protein MERS-CoV, Spike RBD Protein MERS-CoV
    Host:
    Baculovirus-Insect Cells
    Buy from Supplier


    Structured Review

    Sino Biological mers
    <t>AIR</t> assay for antibodies to respiratory viruses. For each antigen, six replicate spots are printed in two different locations on the chip. Each group of six spots is surrounded by negative control reference spots (anti-FITC). Blank (background) areas are included as additional negative controls. Key: 1: human coronavirus (HKU isolate) spike glycoprotein, aa 1–760; 2: <t>MERS-CoV</t> spike glycoprotein, S1 domain; 3: MERS-CoV spike glycoprotein, receptor binding domain (RBD); 4: SARS-CoV spike glycoprotein, S1 domain; 5: SARS-CoV spike glycoprotein, RBD; 6: SARS-CoV-2 spike glycoprotein, S1+S2 ECD; 7: SARS-CoV-2 spike glycoprotein, S2 ECD; 8: SARS-CoV-2 spike glycoprotein, S1 domain; 9: SARS-CoV-2 spike glycoprotein, RBD; 10: human coronavirus (HCoV-229E isolate) spike glycoprotein, S1+S2 ECD; 11: human coronavirus (HCoV-OC43 isolate) spike glycoprotein, S1+S2 ECD; 12: influenza B/Brisbane/2008 hemagglutinin; 13: influenza A/California/2009 (H1N1) hemagglutinin; 14: influenza A/Wisconsin/2005 (H3N2) influenza. F1 , F2 , and F3 are derived from spotting three different dilutions of anti-FITC. The image at right is a representative array exposed to Pooled Normal Human Serum (PNHS) at a 1:4 dilution.
    A DNA sequence encoding the extracellular domain of spike protein MERS CoV AFS88936 1 Met1 Trp1297 was fused with a polyhistidine tag at the C terminus
    https://www.bioz.com/result/mers/product/Sino Biological
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    mers - by Bioz Stars, 2021-05
    86/100 stars

    Images

    1) Product Images from "Array-based analysis of SARS-CoV-2, other coronaviruses, and influenza antibodies in convalescent COVID-19 patients"

    Article Title: Array-based analysis of SARS-CoV-2, other coronaviruses, and influenza antibodies in convalescent COVID-19 patients

    Journal: Biosensors & Bioelectronics

    doi: 10.1016/j.bios.2020.112643

    AIR assay for antibodies to respiratory viruses. For each antigen, six replicate spots are printed in two different locations on the chip. Each group of six spots is surrounded by negative control reference spots (anti-FITC). Blank (background) areas are included as additional negative controls. Key: 1: human coronavirus (HKU isolate) spike glycoprotein, aa 1–760; 2: MERS-CoV spike glycoprotein, S1 domain; 3: MERS-CoV spike glycoprotein, receptor binding domain (RBD); 4: SARS-CoV spike glycoprotein, S1 domain; 5: SARS-CoV spike glycoprotein, RBD; 6: SARS-CoV-2 spike glycoprotein, S1+S2 ECD; 7: SARS-CoV-2 spike glycoprotein, S2 ECD; 8: SARS-CoV-2 spike glycoprotein, S1 domain; 9: SARS-CoV-2 spike glycoprotein, RBD; 10: human coronavirus (HCoV-229E isolate) spike glycoprotein, S1+S2 ECD; 11: human coronavirus (HCoV-OC43 isolate) spike glycoprotein, S1+S2 ECD; 12: influenza B/Brisbane/2008 hemagglutinin; 13: influenza A/California/2009 (H1N1) hemagglutinin; 14: influenza A/Wisconsin/2005 (H3N2) influenza. F1 , F2 , and F3 are derived from spotting three different dilutions of anti-FITC. The image at right is a representative array exposed to Pooled Normal Human Serum (PNHS) at a 1:4 dilution.
    Figure Legend Snippet: AIR assay for antibodies to respiratory viruses. For each antigen, six replicate spots are printed in two different locations on the chip. Each group of six spots is surrounded by negative control reference spots (anti-FITC). Blank (background) areas are included as additional negative controls. Key: 1: human coronavirus (HKU isolate) spike glycoprotein, aa 1–760; 2: MERS-CoV spike glycoprotein, S1 domain; 3: MERS-CoV spike glycoprotein, receptor binding domain (RBD); 4: SARS-CoV spike glycoprotein, S1 domain; 5: SARS-CoV spike glycoprotein, RBD; 6: SARS-CoV-2 spike glycoprotein, S1+S2 ECD; 7: SARS-CoV-2 spike glycoprotein, S2 ECD; 8: SARS-CoV-2 spike glycoprotein, S1 domain; 9: SARS-CoV-2 spike glycoprotein, RBD; 10: human coronavirus (HCoV-229E isolate) spike glycoprotein, S1+S2 ECD; 11: human coronavirus (HCoV-OC43 isolate) spike glycoprotein, S1+S2 ECD; 12: influenza B/Brisbane/2008 hemagglutinin; 13: influenza A/California/2009 (H1N1) hemagglutinin; 14: influenza A/Wisconsin/2005 (H3N2) influenza. F1 , F2 , and F3 are derived from spotting three different dilutions of anti-FITC. The image at right is a representative array exposed to Pooled Normal Human Serum (PNHS) at a 1:4 dilution.

    Techniques Used: Chromatin Immunoprecipitation, Negative Control, Binding Assay, Derivative Assay

    Response of a commercial anti-SARS-CoV-2 rabbit polyclonal antibody (pAb) on the array. (A) Array exposed 20% FBS + 10% PNHS; (B) Array exposed to 1 μg/mL anti-SARS-CoV-2 pAb in 20% FBS + 10% PNHS. Strong responses to SARS-CoV-2 S1+S2 ECD, S1, and RBD are observed, as well as smaller cross-reactive responses to HCoV-229E, HCoV-OC43, and MERS spike proteins; (C) quantitative data for the titration. Concentrations of pAb are provided at the top of each column in ng/mL; response values at each concentration for each antigen are provided in Ångstroms of build. (D) Titration curves for the four SARS-CoV-2 antigens with standard deviation of replicate probe spots at each concentration.
    Figure Legend Snippet: Response of a commercial anti-SARS-CoV-2 rabbit polyclonal antibody (pAb) on the array. (A) Array exposed 20% FBS + 10% PNHS; (B) Array exposed to 1 μg/mL anti-SARS-CoV-2 pAb in 20% FBS + 10% PNHS. Strong responses to SARS-CoV-2 S1+S2 ECD, S1, and RBD are observed, as well as smaller cross-reactive responses to HCoV-229E, HCoV-OC43, and MERS spike proteins; (C) quantitative data for the titration. Concentrations of pAb are provided at the top of each column in ng/mL; response values at each concentration for each antigen are provided in Ångstroms of build. (D) Titration curves for the four SARS-CoV-2 antigens with standard deviation of replicate probe spots at each concentration.

    Techniques Used: Titration, Concentration Assay, Standard Deviation

    2) Product Images from "Construction and next-generation sequencing analysis of a large phage-displayed VNAR single-domain antibody library from six naïve nurse sharks"

    Article Title: Construction and next-generation sequencing analysis of a large phage-displayed VNAR single-domain antibody library from six naïve nurse sharks

    Journal: Antibody Therapeutics

    doi: 10.1093/abt/tby011

    Identification of the binders by phage ELISA. (A) Monoclonal phage ELISA was carried out to identify the binders to GPC3, Her2, PD1, MERS S-protein, SARS S-protein, PE38, and hFc. A random phage that had no binding to all tested antigens was used as irrelevant control in the phage ELISA. (B) Octet kinetic assay for PE38-B6-his single-domain soluble protein. Seven concentrations of PE38-B6-his protein used from high to low are 780 nM, 195 nM, 48.8 nM, 12.2 nM, 3.06 nM, 0.76 nM, and 0.19 nM. (C) The representative data were shown for PE38-B6 concentration 3.06 nM. The K d , k on , k off , and k off standard error was summarized in this table.
    Figure Legend Snippet: Identification of the binders by phage ELISA. (A) Monoclonal phage ELISA was carried out to identify the binders to GPC3, Her2, PD1, MERS S-protein, SARS S-protein, PE38, and hFc. A random phage that had no binding to all tested antigens was used as irrelevant control in the phage ELISA. (B) Octet kinetic assay for PE38-B6-his single-domain soluble protein. Seven concentrations of PE38-B6-his protein used from high to low are 780 nM, 195 nM, 48.8 nM, 12.2 nM, 3.06 nM, 0.76 nM, and 0.19 nM. (C) The representative data were shown for PE38-B6 concentration 3.06 nM. The K d , k on , k off , and k off standard error was summarized in this table.

    Techniques Used: Enzyme-linked Immunosorbent Assay, Binding Assay, Kinetic Assay, Concentration Assay

    3) Product Images from "Array-based analysis of SARS-CoV-2, other coronaviruses, and influenza antibodies in convalescent COVID-19 patients"

    Article Title: Array-based analysis of SARS-CoV-2, other coronaviruses, and influenza antibodies in convalescent COVID-19 patients

    Journal: bioRxiv

    doi: 10.1101/2020.06.15.153064

    AIR assay for antibodies to respiratory viruses. For each antigen, six replicate spots are printed in two different locations on the chip. Each group of six spots is surrounded by negative control reference spots (anti-FITC). Blank (background) areas are included as additional negative controls. Key: 1: human coronavirus (HKU isolate) spike glycoprotein, aa 1-760; 2: MERS-CoV spike glycoprotein, S1 domain; 3: MERS-CoV spike glycoprotein, receptor binding domain (RBD); 4: SARS-CoV spike glycoprotein, S1 domain; 5: SARS-CoV spike glycoprotein, RBD; 6: SARS-CoV-2 spike glycoprotein, S1+S2 ECD; 7: SARS-CoV-2 spike glycoprotein, S2 ECD; 8: SARS-CoV-2 spike glycoprotein, S1 domain; 9: SARS-CoV-2 spike glycoprotein, RBD; 10: human coronavirus (HCoV-229E isolate) spike glycoprotein, S1+S2 ECD; 11: human coronavirus (HCoV-OC43 isolate) spike glycoprotein, S1+S2 ECD; 12: influenza B/Brisbane/2008 hemagglutinin; 13: influenza A/California/2009 (H1N1) hemagglutinin; 14: influenza A/Wisconsin/2005 (H3N2) influenza. F1 , F2 , and F3 are derived from spotting three different dilutions of anti-FITC. The image at right is a representative array exposed to Pooled Normal Human Serum (PNHS) at a 1:4 dilution.
    Figure Legend Snippet: AIR assay for antibodies to respiratory viruses. For each antigen, six replicate spots are printed in two different locations on the chip. Each group of six spots is surrounded by negative control reference spots (anti-FITC). Blank (background) areas are included as additional negative controls. Key: 1: human coronavirus (HKU isolate) spike glycoprotein, aa 1-760; 2: MERS-CoV spike glycoprotein, S1 domain; 3: MERS-CoV spike glycoprotein, receptor binding domain (RBD); 4: SARS-CoV spike glycoprotein, S1 domain; 5: SARS-CoV spike glycoprotein, RBD; 6: SARS-CoV-2 spike glycoprotein, S1+S2 ECD; 7: SARS-CoV-2 spike glycoprotein, S2 ECD; 8: SARS-CoV-2 spike glycoprotein, S1 domain; 9: SARS-CoV-2 spike glycoprotein, RBD; 10: human coronavirus (HCoV-229E isolate) spike glycoprotein, S1+S2 ECD; 11: human coronavirus (HCoV-OC43 isolate) spike glycoprotein, S1+S2 ECD; 12: influenza B/Brisbane/2008 hemagglutinin; 13: influenza A/California/2009 (H1N1) hemagglutinin; 14: influenza A/Wisconsin/2005 (H3N2) influenza. F1 , F2 , and F3 are derived from spotting three different dilutions of anti-FITC. The image at right is a representative array exposed to Pooled Normal Human Serum (PNHS) at a 1:4 dilution.

    Techniques Used: Chromatin Immunoprecipitation, Negative Control, Binding Assay, Derivative Assay

    Response of a commercial anti-SARS-CoV-2 rabbit polyclonal antibody (pAb) on the array. (A) array exposed to array exposed to 20% FBS + 10% PNHS; (B) array exposed to 1 μg/mL anti-SARS-CoV-2 pAb in 20% FBS + 10% PNHS. Strong responses to SARS-CoV-2 S1+S2 ECD, S1, and RBD are observed, as well as smaller cross-reactive responses to HCoV-229E, HCoV-OC43, and MERS spike proteins; (C) quantitative data for the titration. Concentrations of pAb are provided at the top of each column in ng/mL; response values at each concentration for each antigen are provided in Angstroms of build. (D) Titration curves for the four SARS-CoV-2 antigens with standard deviation of replicate probe spots at each concentration.
    Figure Legend Snippet: Response of a commercial anti-SARS-CoV-2 rabbit polyclonal antibody (pAb) on the array. (A) array exposed to array exposed to 20% FBS + 10% PNHS; (B) array exposed to 1 μg/mL anti-SARS-CoV-2 pAb in 20% FBS + 10% PNHS. Strong responses to SARS-CoV-2 S1+S2 ECD, S1, and RBD are observed, as well as smaller cross-reactive responses to HCoV-229E, HCoV-OC43, and MERS spike proteins; (C) quantitative data for the titration. Concentrations of pAb are provided at the top of each column in ng/mL; response values at each concentration for each antigen are provided in Angstroms of build. (D) Titration curves for the four SARS-CoV-2 antigens with standard deviation of replicate probe spots at each concentration.

    Techniques Used: Titration, Concentration Assay, Standard Deviation

    4) Product Images from "Construction and next-generation sequencing analysis of a large phage-displayed VNAR single-domain antibody library from six naïve nurse sharks"

    Article Title: Construction and next-generation sequencing analysis of a large phage-displayed VNAR single-domain antibody library from six naïve nurse sharks

    Journal: Antibody Therapeutics

    doi: 10.1093/abt/tby011

    Identification of the binders by phage ELISA. (A) Monoclonal phage ELISA was carried out to identify the binders to GPC3, Her2, PD1, MERS S-protein, SARS S-protein, PE38, and hFc. A random phage that had no binding to all tested antigens was used as irrelevant control in the phage ELISA. (B) Octet kinetic assay for PE38-B6-his single-domain soluble protein. Seven concentrations of PE38-B6-his protein used from high to low are 780 nM, 195 nM, 48.8 nM, 12.2 nM, 3.06 nM, 0.76 nM, and 0.19 nM. (C) The representative data were shown for PE38-B6 concentration 3.06 nM. The K d , k on , k off , and k off standard error was summarized in this table.
    Figure Legend Snippet: Identification of the binders by phage ELISA. (A) Monoclonal phage ELISA was carried out to identify the binders to GPC3, Her2, PD1, MERS S-protein, SARS S-protein, PE38, and hFc. A random phage that had no binding to all tested antigens was used as irrelevant control in the phage ELISA. (B) Octet kinetic assay for PE38-B6-his single-domain soluble protein. Seven concentrations of PE38-B6-his protein used from high to low are 780 nM, 195 nM, 48.8 nM, 12.2 nM, 3.06 nM, 0.76 nM, and 0.19 nM. (C) The representative data were shown for PE38-B6 concentration 3.06 nM. The K d , k on , k off , and k off standard error was summarized in this table.

    Techniques Used: Enzyme-linked Immunosorbent Assay, Binding Assay, Kinetic Assay, Concentration Assay

    Related Articles

    Infection:

    Article Title: Inhibition of Proprotein Convertases Abrogates Processing of the Middle Eastern Respiratory Syndrome Coronavirus Spike Protein in Infected Cells but Does Not Reduce Viral Infectivity
    Article Snippet: MERS-S expression was detected using a monoclonal antibody directed against the V5 tag (Invitrogen) or a polyclonal antibody directed against the S2 subunit of the MERS-S protein (Sino Biological). .. For detection of MERS-S protein in infected cells, Caco-2 and Vero B4 cells were infected with MERS-CoV (human betacoronavirus 2c EMC/2012) at a multiplicity of infection (MOI) of 0.01 and 5, respectively. .. At 24 hours after infection, the cells were harvested and treated with NuPAGE LDS sample buffer (Invitrogen), boiled for 20 minutes at 95°C, and analyzed by Western blot as described above.

    Incubation:

    Article Title: HTCC as a Polymeric Inhibitor of SARS-CoV-2 and MERS-CoV
    Article Snippet: Immunostaining and confocal imaging.Fixed cells were permeabilized with 0.1% Triton X-100–1× PBS and incubated overnight at 4°C in 1× PBS supplemented with 5% bovine serum albumin (BSA) and 0.5% Tween 20. .. To visualize MERS-CoV particles, cells were incubated for 2 h at room temperature with mouse anti-MERS-CoV N IgGs (Sino Biological, China) (1:1,000 dilution), followed by 1 h of incubation with Alexa Fluor 488-labeled goat anti-mouse IgG (Thermo Fisher Scientific, Poland) (2.5 μg/ml). .. Actin filaments were stained using phalloidin coupled with Alexa Fluor 633 (Thermo Fisher Scientific, Poland) (0.2 U/ml).

    other:

    Article Title: Safety and immunogenicity of a candidate Middle East respiratory syndrome coronavirus viral-vectored vaccine: a dose-escalation, open-label, non-randomised, uncontrolled, phase 1 trial
    Article Snippet: Peptides were pooled into 13 pools for the MERS-CoV spike protein containing 18 or 21 peptides, plus a single pool of five peptides for the tissue plasminogen activator leader.

    Article Title: Ultrapotent Human Neutralizing Antibody Repertoires Against Middle East Respiratory Syndrome Coronavirus From a Recovered Patient
    Article Snippet: MERS-GD27 and MERS-GD33 showed subnanomolar affinity for the MERS-CoV S protein (K D equivalent to 0.775 and 0.575 nM, respectively), which was consistent with MERS-4 (K D equivalent to 0.978 nM) [ ].

    Article Title: A unifying structural and functional model of the coronavirus replication organelle: tracking down RNA synthesis
    Article Snippet: The human monoclonal antibody used against MERS-CoV spike protein was kindly provided by Dr. Berend Jan Bosch (Utrecht University) and has been described elsewhere as 1.6f9 [ ].

    Enzyme-linked Immunosorbent Assay:

    Article Title: High Prevalence of Middle East Respiratory Coronavirus in Young Dromedary Camels in Jordan
    Article Snippet: Phylogenetic trees of the S2 domain were generated using Mega 6.0.6 with the maximum likelihood statistical method based on the GTR+G+I model with 1000 bootstraps replicates. .. Sera were analyzed by MERS-CoV spike protein (S) enzyme-linked immunosorbent assay (ELISA); Maxisorp (Nunc) plates were coated overnight with S1 protein (Sino Biological) and blocked with 1% milk. ..

    Functional Assay:

    Article Title: Ultrapotent Human Neutralizing Antibody Repertoires Against Middle East Respiratory Syndrome Coronavirus From a Recovered Patient
    Article Snippet: .. Functional Characterization of Antibodies With Potent Neutralizing Activity Against Middle East Respiratory Syndrome Coronavirus In VitroTo characterize the function of human mAbs targeting the MERS-CoV S protein, we produced 13 mAbs from 11 cell cultures in large quantities from different combinations of VH and VL/VK genes in the same well by a double gene expression vector transiently transfected into human embryonic kidney (HEK) 293FS cells. .. All of the Abs target the S protein from the endoplasmic reticulum membrane protein complex (EMC) strain, as determined by ELISA, whereas the H7 (a mAb against hemagglutination of the influenza virus) was used as an irrelevant Ab control and phosphate-buffered saline was used as a blank control ( ).

    Activity Assay:

    Article Title: Ultrapotent Human Neutralizing Antibody Repertoires Against Middle East Respiratory Syndrome Coronavirus From a Recovered Patient
    Article Snippet: .. Functional Characterization of Antibodies With Potent Neutralizing Activity Against Middle East Respiratory Syndrome Coronavirus In VitroTo characterize the function of human mAbs targeting the MERS-CoV S protein, we produced 13 mAbs from 11 cell cultures in large quantities from different combinations of VH and VL/VK genes in the same well by a double gene expression vector transiently transfected into human embryonic kidney (HEK) 293FS cells. .. All of the Abs target the S protein from the endoplasmic reticulum membrane protein complex (EMC) strain, as determined by ELISA, whereas the H7 (a mAb against hemagglutination of the influenza virus) was used as an irrelevant Ab control and phosphate-buffered saline was used as a blank control ( ).

    Produced:

    Article Title: Ultrapotent Human Neutralizing Antibody Repertoires Against Middle East Respiratory Syndrome Coronavirus From a Recovered Patient
    Article Snippet: .. Functional Characterization of Antibodies With Potent Neutralizing Activity Against Middle East Respiratory Syndrome Coronavirus In VitroTo characterize the function of human mAbs targeting the MERS-CoV S protein, we produced 13 mAbs from 11 cell cultures in large quantities from different combinations of VH and VL/VK genes in the same well by a double gene expression vector transiently transfected into human embryonic kidney (HEK) 293FS cells. .. All of the Abs target the S protein from the endoplasmic reticulum membrane protein complex (EMC) strain, as determined by ELISA, whereas the H7 (a mAb against hemagglutination of the influenza virus) was used as an irrelevant Ab control and phosphate-buffered saline was used as a blank control ( ).

    Expressing:

    Article Title: Ultrapotent Human Neutralizing Antibody Repertoires Against Middle East Respiratory Syndrome Coronavirus From a Recovered Patient
    Article Snippet: .. Functional Characterization of Antibodies With Potent Neutralizing Activity Against Middle East Respiratory Syndrome Coronavirus In VitroTo characterize the function of human mAbs targeting the MERS-CoV S protein, we produced 13 mAbs from 11 cell cultures in large quantities from different combinations of VH and VL/VK genes in the same well by a double gene expression vector transiently transfected into human embryonic kidney (HEK) 293FS cells. .. All of the Abs target the S protein from the endoplasmic reticulum membrane protein complex (EMC) strain, as determined by ELISA, whereas the H7 (a mAb against hemagglutination of the influenza virus) was used as an irrelevant Ab control and phosphate-buffered saline was used as a blank control ( ).

    Plasmid Preparation:

    Article Title: Ultrapotent Human Neutralizing Antibody Repertoires Against Middle East Respiratory Syndrome Coronavirus From a Recovered Patient
    Article Snippet: .. Functional Characterization of Antibodies With Potent Neutralizing Activity Against Middle East Respiratory Syndrome Coronavirus In VitroTo characterize the function of human mAbs targeting the MERS-CoV S protein, we produced 13 mAbs from 11 cell cultures in large quantities from different combinations of VH and VL/VK genes in the same well by a double gene expression vector transiently transfected into human embryonic kidney (HEK) 293FS cells. .. All of the Abs target the S protein from the endoplasmic reticulum membrane protein complex (EMC) strain, as determined by ELISA, whereas the H7 (a mAb against hemagglutination of the influenza virus) was used as an irrelevant Ab control and phosphate-buffered saline was used as a blank control ( ).

    Transfection:

    Article Title: Ultrapotent Human Neutralizing Antibody Repertoires Against Middle East Respiratory Syndrome Coronavirus From a Recovered Patient
    Article Snippet: .. Functional Characterization of Antibodies With Potent Neutralizing Activity Against Middle East Respiratory Syndrome Coronavirus In VitroTo characterize the function of human mAbs targeting the MERS-CoV S protein, we produced 13 mAbs from 11 cell cultures in large quantities from different combinations of VH and VL/VK genes in the same well by a double gene expression vector transiently transfected into human embryonic kidney (HEK) 293FS cells. .. All of the Abs target the S protein from the endoplasmic reticulum membrane protein complex (EMC) strain, as determined by ELISA, whereas the H7 (a mAb against hemagglutination of the influenza virus) was used as an irrelevant Ab control and phosphate-buffered saline was used as a blank control ( ).

    Similar Products

  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 96
    Sino Biological mers cov spike protein
    Single-dose intranasal immunization with <t>PIV5-MERS-S</t> induced robust <t>MERS-CoV-specific</t> CD8 T cell response in human DPP4 knockin (hDPP4 KI) mice. (A) Schematic diagram showing the experimental plan to examine CD8 T cell response after immunization and challenge. hDPP4 KI mice were intranasally immunized with 10 4 PFU PIV5-MERS-S or PIV5-GFP. At 4 weeks or 4 days following immunization, mice were challenged with MERS MA 6.1.2, single-cell suspensions from the lungs of immunized mice were stimulated with MERS-CoV spike peptides (S343 and S1165), and specific CD8 T cells were determined by IFN-γ intracellular staining. (B to D) Representative FACS plots (B), percentage (C), and total number (D) of MERS-CoV-specific CD8 T cells in the lungs at 4 weeks after immunization. (E to G) Representative FACS plots (E), percentage (F), and total number (G) of MERS-CoV-specific CD8 T cells in the lungs at 4 days after MERS MA 6.1.2 challenge ( n = 6 mice per group). Data are representative of two independent experiments. Data presented represent mean ± SE; * denotes P
    Mers Cov Spike Protein, supplied by Sino Biological, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/mers cov spike protein/product/Sino Biological
    Average 96 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    mers cov spike protein - by Bioz Stars, 2021-05
    96/100 stars
      Buy from Supplier

    97
    Sino Biological mouse anti mers cov n iggs
    Coronavirus internalization into susceptible cells is hampered by HTCC. (A and B) Precooled HAE cultures were incubated with an ice-cold <t>MERS-CoV</t> suspension in the presence or absence of HTCC-63 (200 μg/ml) for 2 h at 37°C. Next, cells were fixed in paraformaldehyde (PFA) and immunostained for MERS-CoV N protein and actin. Virus entry was analyzed with confocal microscopy. The data shown are representative of results from three independent experiments, each performed in triplicate. Mann-Whitney test, *** * , P
    Mouse Anti Mers Cov N Iggs, supplied by Sino Biological, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/mouse anti mers cov n iggs/product/Sino Biological
    Average 97 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    mouse anti mers cov n iggs - by Bioz Stars, 2021-05
    97/100 stars
      Buy from Supplier

    96
    Sino Biological rabbit anti mers cov antibodies
    Characterization of <t>anti-MERS-S2P</t> IgGs. ( A ) Immunofluorescence assay of anti-MERS-S2P IgGs on cell lines infected with various human coronaviruses (hCoVs) to determine their <t>MERS-CoV</t> specificity. Scale bar, 200 μm. ( B ) Size-exclusion chromatography analysis of S2A3 (IgG) and S2D5 (IgG). The molecular weights (kDa) of the molecular mass markers are shown above the corresponding peaks at the top chromatogram. ( C ) Surface plasmon resonance (SPR) analysis of S2A3 (IgG) and S2D5 (IgG) on a MERS-S2P-immobilized sensor chip to determine their apparent binding affinities. The fitted-lines are marked by red.
    Rabbit Anti Mers Cov Antibodies, supplied by Sino Biological, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rabbit anti mers cov antibodies/product/Sino Biological
    Average 96 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    rabbit anti mers cov antibodies - by Bioz Stars, 2021-05
    96/100 stars
      Buy from Supplier

    93
    Sino Biological monoclonal mouse anti mers cov nucleocapsid antibody
    Mapping of H2-d restricted T cell epitopes in <t>MERS-CoV</t> N protein; ( a–b ) BALB/c mice (n = 2 to 4) were immunized twice (21-day interval) i.p. or i.m. with 10 8 PFU of recombinant MVA-MERS-N (MVA-N), non-recombinant MVA (MVA) or PBS. Splenocytes from vaccinated mice were incubated in the presence of subpools (V8.1, V8.2, H8.1, H8.2) from positive matrix pools ( a ) or individual 15-mers peptides #89 or #90 ( b ). IFN-γ spot-forming CD8+ T cells (IFN-γ SFC) were quantified by ELISPOT. The lines represent means.
    Monoclonal Mouse Anti Mers Cov Nucleocapsid Antibody, supplied by Sino Biological, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/monoclonal mouse anti mers cov nucleocapsid antibody/product/Sino Biological
    Average 93 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    monoclonal mouse anti mers cov nucleocapsid antibody - by Bioz Stars, 2021-05
    93/100 stars
      Buy from Supplier

    Image Search Results


    Single-dose intranasal immunization with PIV5-MERS-S induced robust MERS-CoV-specific CD8 T cell response in human DPP4 knockin (hDPP4 KI) mice. (A) Schematic diagram showing the experimental plan to examine CD8 T cell response after immunization and challenge. hDPP4 KI mice were intranasally immunized with 10 4 PFU PIV5-MERS-S or PIV5-GFP. At 4 weeks or 4 days following immunization, mice were challenged with MERS MA 6.1.2, single-cell suspensions from the lungs of immunized mice were stimulated with MERS-CoV spike peptides (S343 and S1165), and specific CD8 T cells were determined by IFN-γ intracellular staining. (B to D) Representative FACS plots (B), percentage (C), and total number (D) of MERS-CoV-specific CD8 T cells in the lungs at 4 weeks after immunization. (E to G) Representative FACS plots (E), percentage (F), and total number (G) of MERS-CoV-specific CD8 T cells in the lungs at 4 days after MERS MA 6.1.2 challenge ( n = 6 mice per group). Data are representative of two independent experiments. Data presented represent mean ± SE; * denotes P

    Journal: mBio

    Article Title: Single-Dose, Intranasal Immunization with Recombinant Parainfluenza Virus 5 Expressing Middle East Respiratory Syndrome Coronavirus (MERS-CoV) Spike Protein Protects Mice from Fatal MERS-CoV Infection

    doi: 10.1128/mBio.00554-20

    Figure Lengend Snippet: Single-dose intranasal immunization with PIV5-MERS-S induced robust MERS-CoV-specific CD8 T cell response in human DPP4 knockin (hDPP4 KI) mice. (A) Schematic diagram showing the experimental plan to examine CD8 T cell response after immunization and challenge. hDPP4 KI mice were intranasally immunized with 10 4 PFU PIV5-MERS-S or PIV5-GFP. At 4 weeks or 4 days following immunization, mice were challenged with MERS MA 6.1.2, single-cell suspensions from the lungs of immunized mice were stimulated with MERS-CoV spike peptides (S343 and S1165), and specific CD8 T cells were determined by IFN-γ intracellular staining. (B to D) Representative FACS plots (B), percentage (C), and total number (D) of MERS-CoV-specific CD8 T cells in the lungs at 4 weeks after immunization. (E to G) Representative FACS plots (E), percentage (F), and total number (G) of MERS-CoV-specific CD8 T cells in the lungs at 4 days after MERS MA 6.1.2 challenge ( n = 6 mice per group). Data are representative of two independent experiments. Data presented represent mean ± SE; * denotes P

    Article Snippet: The expression of MERS-CoV spike protein was detected by a rabbit anti-MERS-CoV S2 antibody (Sino Biological, catalog no. 40070-T60) and colocalized using a mouse anti-α-tubulin antibody (Sigma, catalog no. T9026).

    Techniques: Knock-In, Mouse Assay, Staining, FACS

    Serum neutralizing antibodies produced in mice 4 weeks after single-dose intranasal immunization with PIV5-MERS-S. Naive mice were intranasally immunized with PIV5-GFP or PIV5-MERS-S. Sera were collected at 4 weeks postimmunization. Neutralization assay against MERS-CoV spike pseudovirions was performed as described in Materials and Methods. The neutralization results were measured in luciferase activity and plotted relative to mock-treatment value. (A and B) Neutralization assay results from C57BL/6 mice immunized with 10 4 PFU (A) and 10 6 PFU (B) PIV5-MERS-S or PIV5-GFP. (C) Neutralization assay results from BALB/c mice immunized with 10 6 PFU PIV5-MERS-S or PIV5-GFP. Data presented represent means ± SEs.

    Journal: mBio

    Article Title: Single-Dose, Intranasal Immunization with Recombinant Parainfluenza Virus 5 Expressing Middle East Respiratory Syndrome Coronavirus (MERS-CoV) Spike Protein Protects Mice from Fatal MERS-CoV Infection

    doi: 10.1128/mBio.00554-20

    Figure Lengend Snippet: Serum neutralizing antibodies produced in mice 4 weeks after single-dose intranasal immunization with PIV5-MERS-S. Naive mice were intranasally immunized with PIV5-GFP or PIV5-MERS-S. Sera were collected at 4 weeks postimmunization. Neutralization assay against MERS-CoV spike pseudovirions was performed as described in Materials and Methods. The neutralization results were measured in luciferase activity and plotted relative to mock-treatment value. (A and B) Neutralization assay results from C57BL/6 mice immunized with 10 4 PFU (A) and 10 6 PFU (B) PIV5-MERS-S or PIV5-GFP. (C) Neutralization assay results from BALB/c mice immunized with 10 6 PFU PIV5-MERS-S or PIV5-GFP. Data presented represent means ± SEs.

    Article Snippet: The expression of MERS-CoV spike protein was detected by a rabbit anti-MERS-CoV S2 antibody (Sino Biological, catalog no. 40070-T60) and colocalized using a mouse anti-α-tubulin antibody (Sigma, catalog no. T9026).

    Techniques: Produced, Mouse Assay, Neutralization, Luciferase, Activity Assay

    Representative images of lung tissues from mice receiving PBS (A), UV-inactivated MERS-CoV (B), or PIV5-MERS-S (C) treatment, followed by infection with MERS MA 6.1.2. Images obtained from tissues at 3 days after MERS MA 6.1.2 infection. Compared to PBS or PIV5-MERS, perivascular eosinophilic infiltration (arrows) in UV-MERS-treated mice was greatly increased. n = 3 to 4 mice/group. (D) Graph representing eosinophil infiltration in the lung tissues of mice from groups shown in panels A to . * denotes P

    Journal: mBio

    Article Title: Single-Dose, Intranasal Immunization with Recombinant Parainfluenza Virus 5 Expressing Middle East Respiratory Syndrome Coronavirus (MERS-CoV) Spike Protein Protects Mice from Fatal MERS-CoV Infection

    doi: 10.1128/mBio.00554-20

    Figure Lengend Snippet: Representative images of lung tissues from mice receiving PBS (A), UV-inactivated MERS-CoV (B), or PIV5-MERS-S (C) treatment, followed by infection with MERS MA 6.1.2. Images obtained from tissues at 3 days after MERS MA 6.1.2 infection. Compared to PBS or PIV5-MERS, perivascular eosinophilic infiltration (arrows) in UV-MERS-treated mice was greatly increased. n = 3 to 4 mice/group. (D) Graph representing eosinophil infiltration in the lung tissues of mice from groups shown in panels A to . * denotes P

    Article Snippet: The expression of MERS-CoV spike protein was detected by a rabbit anti-MERS-CoV S2 antibody (Sino Biological, catalog no. 40070-T60) and colocalized using a mouse anti-α-tubulin antibody (Sigma, catalog no. T9026).

    Techniques: Mouse Assay, Infection

    Histopathology in immunized mice challenged with MERS-CoV. hDPP4 KI mice were intranasally immunized with 10 4 PFU PIV5-MERS-S or PIV5-GFP. Four weeks after immunization, the mice were intranasally infected with 10 5 PFU MERS MA 6.1.2. (A) Representative images of H E-stained lung sections from PIV5-MERS-S- or PIV5-GFP-immunized hDPP4 KI mice at indicated days after MERS MA 6.1.2 challenge. Note the cellular infiltration (black arrows) and the hyaline membranes (red arrows). (B) Summary scores for disease in the lung sections. n = 3 to 5 mice/group. * denotes P

    Journal: mBio

    Article Title: Single-Dose, Intranasal Immunization with Recombinant Parainfluenza Virus 5 Expressing Middle East Respiratory Syndrome Coronavirus (MERS-CoV) Spike Protein Protects Mice from Fatal MERS-CoV Infection

    doi: 10.1128/mBio.00554-20

    Figure Lengend Snippet: Histopathology in immunized mice challenged with MERS-CoV. hDPP4 KI mice were intranasally immunized with 10 4 PFU PIV5-MERS-S or PIV5-GFP. Four weeks after immunization, the mice were intranasally infected with 10 5 PFU MERS MA 6.1.2. (A) Representative images of H E-stained lung sections from PIV5-MERS-S- or PIV5-GFP-immunized hDPP4 KI mice at indicated days after MERS MA 6.1.2 challenge. Note the cellular infiltration (black arrows) and the hyaline membranes (red arrows). (B) Summary scores for disease in the lung sections. n = 3 to 5 mice/group. * denotes P

    Article Snippet: The expression of MERS-CoV spike protein was detected by a rabbit anti-MERS-CoV S2 antibody (Sino Biological, catalog no. 40070-T60) and colocalized using a mouse anti-α-tubulin antibody (Sigma, catalog no. T9026).

    Techniques: Histopathology, Mouse Assay, Infection, Staining

    Comparison of the protective efficacy between single-dose immunization with UV light-inactivated MERS-CoV and PIV5-MERS-S. hDPP4 KI mice were immunized with 10 4 PFU PIV5-MERS-S via intranasal route; 10 6 PFU UV-inactivated MERS MA 6.1.2, mixed with Imject alum; or PBS via intramuscular route. Four weeks after immunization, immunized mice were infected with 10 5 PFU MERS-CoV. (A) Schematic timeline outlining experimental plan. (B and C) Survival (B) and weight loss (C) were monitored daily until 10 days postinfection. PBS, n = 9; UV MERS-CoV, n = 12; PIV5-MERS-S, n = 8. Data represent mean ± SE.

    Journal: mBio

    Article Title: Single-Dose, Intranasal Immunization with Recombinant Parainfluenza Virus 5 Expressing Middle East Respiratory Syndrome Coronavirus (MERS-CoV) Spike Protein Protects Mice from Fatal MERS-CoV Infection

    doi: 10.1128/mBio.00554-20

    Figure Lengend Snippet: Comparison of the protective efficacy between single-dose immunization with UV light-inactivated MERS-CoV and PIV5-MERS-S. hDPP4 KI mice were immunized with 10 4 PFU PIV5-MERS-S via intranasal route; 10 6 PFU UV-inactivated MERS MA 6.1.2, mixed with Imject alum; or PBS via intramuscular route. Four weeks after immunization, immunized mice were infected with 10 5 PFU MERS-CoV. (A) Schematic timeline outlining experimental plan. (B and C) Survival (B) and weight loss (C) were monitored daily until 10 days postinfection. PBS, n = 9; UV MERS-CoV, n = 12; PIV5-MERS-S, n = 8. Data represent mean ± SE.

    Article Snippet: The expression of MERS-CoV spike protein was detected by a rabbit anti-MERS-CoV S2 antibody (Sino Biological, catalog no. 40070-T60) and colocalized using a mouse anti-α-tubulin antibody (Sigma, catalog no. T9026).

    Techniques: Mouse Assay, Infection

    Generation and characterization of recombinant PIV5 expressing MERS-CoV spike protein. (A) Schematic of PIV5-MERS-S. NP, nucleoprotein; V, V protein; P, phosphoprotein; M, matrix protein; F, fusion protein; SH, small hydrophobic protein; HN, hemagglutinin-neuraminidase protein; L, RNA-dependent RNA polymerase. (B) Confirmation of MERS-CoV spike protein expression by Western blotting. Vero 81 cells were infected with PIV5-MERS-S at MOIs of 0.01, 0.1, and 1.0 or mock infected. At 2 days postinfection, MERS-CoV spike was detected with anti-MERS-S antibody by Western blotting. (C) Immunofluorescence of Vero cells infected with PIV5 and PIV5-MERS-S. Vero cells were infected with PIV5 and PIV5-MERS-S (MOI = 0.1) or mock infected. At 2 days postinfection, cells were fixed, permeabilized, and stained with anti-PIV5 V/P or anti-MERS-spike antibodies. Scale bar = 200 μm. (D) Growth rate of PIV5-MERS-S. MDBK cells were infected with PIV5 or PIV5-MERS-S at an MOI of 0.1. Media were collected daily for 5 days, and titers of viruses in the media were determined using plaque assay.

    Journal: mBio

    Article Title: Single-Dose, Intranasal Immunization with Recombinant Parainfluenza Virus 5 Expressing Middle East Respiratory Syndrome Coronavirus (MERS-CoV) Spike Protein Protects Mice from Fatal MERS-CoV Infection

    doi: 10.1128/mBio.00554-20

    Figure Lengend Snippet: Generation and characterization of recombinant PIV5 expressing MERS-CoV spike protein. (A) Schematic of PIV5-MERS-S. NP, nucleoprotein; V, V protein; P, phosphoprotein; M, matrix protein; F, fusion protein; SH, small hydrophobic protein; HN, hemagglutinin-neuraminidase protein; L, RNA-dependent RNA polymerase. (B) Confirmation of MERS-CoV spike protein expression by Western blotting. Vero 81 cells were infected with PIV5-MERS-S at MOIs of 0.01, 0.1, and 1.0 or mock infected. At 2 days postinfection, MERS-CoV spike was detected with anti-MERS-S antibody by Western blotting. (C) Immunofluorescence of Vero cells infected with PIV5 and PIV5-MERS-S. Vero cells were infected with PIV5 and PIV5-MERS-S (MOI = 0.1) or mock infected. At 2 days postinfection, cells were fixed, permeabilized, and stained with anti-PIV5 V/P or anti-MERS-spike antibodies. Scale bar = 200 μm. (D) Growth rate of PIV5-MERS-S. MDBK cells were infected with PIV5 or PIV5-MERS-S at an MOI of 0.1. Media were collected daily for 5 days, and titers of viruses in the media were determined using plaque assay.

    Article Snippet: The expression of MERS-CoV spike protein was detected by a rabbit anti-MERS-CoV S2 antibody (Sino Biological, catalog no. 40070-T60) and colocalized using a mouse anti-α-tubulin antibody (Sigma, catalog no. T9026).

    Techniques: Recombinant, Expressing, Western Blot, Infection, Immunofluorescence, Staining, Plaque Assay

    Single-dose intranasal immunization with PIV5-MERS-S completely protects hDPP4 KI mice from lethal MERS-CoV challenge. (A) Schematic timeline showing immunization, challenge, and the evaluation of protection. hDPP4 KI mice were intranasally immunized with 10 4 PFU PIV5-MERS-S, PIV5-GFP, or PBS. Four weeks after immunization, the mice were intranasally infected with 10 5 PFU MERS MA 6.1.2. (B and C) Survival (B) and weight loss (C) were monitored daily for 12 days. PIV5-MERS-S or PIV5-GFP, n = 10; PBS, n = 5. (D) At indicated days postinfection, virus lung titers were quantified by plaque assay. Data are representative of two independent experiments. Data presented represent mean ± SE; * denotes P

    Journal: mBio

    Article Title: Single-Dose, Intranasal Immunization with Recombinant Parainfluenza Virus 5 Expressing Middle East Respiratory Syndrome Coronavirus (MERS-CoV) Spike Protein Protects Mice from Fatal MERS-CoV Infection

    doi: 10.1128/mBio.00554-20

    Figure Lengend Snippet: Single-dose intranasal immunization with PIV5-MERS-S completely protects hDPP4 KI mice from lethal MERS-CoV challenge. (A) Schematic timeline showing immunization, challenge, and the evaluation of protection. hDPP4 KI mice were intranasally immunized with 10 4 PFU PIV5-MERS-S, PIV5-GFP, or PBS. Four weeks after immunization, the mice were intranasally infected with 10 5 PFU MERS MA 6.1.2. (B and C) Survival (B) and weight loss (C) were monitored daily for 12 days. PIV5-MERS-S or PIV5-GFP, n = 10; PBS, n = 5. (D) At indicated days postinfection, virus lung titers were quantified by plaque assay. Data are representative of two independent experiments. Data presented represent mean ± SE; * denotes P

    Article Snippet: The expression of MERS-CoV spike protein was detected by a rabbit anti-MERS-CoV S2 antibody (Sino Biological, catalog no. 40070-T60) and colocalized using a mouse anti-α-tubulin antibody (Sigma, catalog no. T9026).

    Techniques: Mouse Assay, Infection, Plaque Assay

    Coronavirus internalization into susceptible cells is hampered by HTCC. (A and B) Precooled HAE cultures were incubated with an ice-cold MERS-CoV suspension in the presence or absence of HTCC-63 (200 μg/ml) for 2 h at 37°C. Next, cells were fixed in paraformaldehyde (PFA) and immunostained for MERS-CoV N protein and actin. Virus entry was analyzed with confocal microscopy. The data shown are representative of results from three independent experiments, each performed in triplicate. Mann-Whitney test, *** * , P

    Journal: Journal of Virology

    Article Title: HTCC as a Polymeric Inhibitor of SARS-CoV-2 and MERS-CoV

    doi: 10.1128/JVI.01622-20

    Figure Lengend Snippet: Coronavirus internalization into susceptible cells is hampered by HTCC. (A and B) Precooled HAE cultures were incubated with an ice-cold MERS-CoV suspension in the presence or absence of HTCC-63 (200 μg/ml) for 2 h at 37°C. Next, cells were fixed in paraformaldehyde (PFA) and immunostained for MERS-CoV N protein and actin. Virus entry was analyzed with confocal microscopy. The data shown are representative of results from three independent experiments, each performed in triplicate. Mann-Whitney test, *** * , P

    Article Snippet: Mouse anti-MERS-CoV N IgGs (Sino Biological, China) (1:1,000 dilution) and horseradish peroxidase (HRP)-labeled rabbit anti-mouse IgG (Dako, Denmark) (65 ng/ml) were used to detect MERS-CoV nucleocapsid protein.

    Techniques: Incubation, Confocal Microscopy, MANN-WHITNEY

    HTCC blocks the interaction between the virus and its entry receptor. Precooled Huh7 cells were incubated for 3 h at 4°C with ice-cold MERS-CoV or subjected to mock treatment in the presence or absence of HTCC-63 (100 μg/ml). Next, cells were fixed with PFA and immunostained for MERS-CoV-N (green), DPP4 (red), and nuclear DNA (blue). MERS-CoV interaction with the DPP4 protein was analyzed with confocal microscopy. Colocalization of DPP4 with MERS-CoV-N was determined by confocal microscopy, and the results are presented as Manders’ M2 coefficient values after excluding nonspecific nuclear signal. Colocalization analysis was carried out with ImageJ JACoP (Just Another Colocalization Plugin). The decrease in colocalization was statistically significant ( P

    Journal: Journal of Virology

    Article Title: HTCC as a Polymeric Inhibitor of SARS-CoV-2 and MERS-CoV

    doi: 10.1128/JVI.01622-20

    Figure Lengend Snippet: HTCC blocks the interaction between the virus and its entry receptor. Precooled Huh7 cells were incubated for 3 h at 4°C with ice-cold MERS-CoV or subjected to mock treatment in the presence or absence of HTCC-63 (100 μg/ml). Next, cells were fixed with PFA and immunostained for MERS-CoV-N (green), DPP4 (red), and nuclear DNA (blue). MERS-CoV interaction with the DPP4 protein was analyzed with confocal microscopy. Colocalization of DPP4 with MERS-CoV-N was determined by confocal microscopy, and the results are presented as Manders’ M2 coefficient values after excluding nonspecific nuclear signal. Colocalization analysis was carried out with ImageJ JACoP (Just Another Colocalization Plugin). The decrease in colocalization was statistically significant ( P

    Article Snippet: Mouse anti-MERS-CoV N IgGs (Sino Biological, China) (1:1,000 dilution) and horseradish peroxidase (HRP)-labeled rabbit anti-mouse IgG (Dako, Denmark) (65 ng/ml) were used to detect MERS-CoV nucleocapsid protein.

    Techniques: Incubation, Confocal Microscopy

    Characterization of anti-MERS-S2P IgGs. ( A ) Immunofluorescence assay of anti-MERS-S2P IgGs on cell lines infected with various human coronaviruses (hCoVs) to determine their MERS-CoV specificity. Scale bar, 200 μm. ( B ) Size-exclusion chromatography analysis of S2A3 (IgG) and S2D5 (IgG). The molecular weights (kDa) of the molecular mass markers are shown above the corresponding peaks at the top chromatogram. ( C ) Surface plasmon resonance (SPR) analysis of S2A3 (IgG) and S2D5 (IgG) on a MERS-S2P-immobilized sensor chip to determine their apparent binding affinities. The fitted-lines are marked by red.

    Journal: Antibodies

    Article Title: Selection and Characterization of Monoclonal Antibodies Targeting Middle East Respiratory Syndrome Coronavirus through a Human Synthetic Fab Phage Display Library Panning

    doi: 10.3390/antib8030042

    Figure Lengend Snippet: Characterization of anti-MERS-S2P IgGs. ( A ) Immunofluorescence assay of anti-MERS-S2P IgGs on cell lines infected with various human coronaviruses (hCoVs) to determine their MERS-CoV specificity. Scale bar, 200 μm. ( B ) Size-exclusion chromatography analysis of S2A3 (IgG) and S2D5 (IgG). The molecular weights (kDa) of the molecular mass markers are shown above the corresponding peaks at the top chromatogram. ( C ) Surface plasmon resonance (SPR) analysis of S2A3 (IgG) and S2D5 (IgG) on a MERS-S2P-immobilized sensor chip to determine their apparent binding affinities. The fitted-lines are marked by red.

    Article Snippet: The rabbit anti-MERS-CoV antibodies (1:3000, Sino Biological), serially two-fold-diluted MERS-S2P, and HRP-conjugated goat anti-rabbit antibodies (1:6000, Sigma Aldrich) were added and incubated for 30 min at RT.

    Techniques: Immunofluorescence, Infection, Size-exclusion Chromatography, SPR Assay, Chromatin Immunoprecipitation, Binding Assay

    Output of the panning of the phage-displayed synthetic Fab library on MERS-S2P. ( A ) Monitoring of phage titers over three rounds (R1–R3) of panning. Black and gray bars indicate a ratio of phage output to input titers presented as a percentage (%) from panning on MERS-S2P-immobilized and -non-immobilized surfaces, respectively. The ratio of output to input (%) = (phage output titer ÷ phage input titer) × 100. ( B ) Phage ELISAs performed on MERS-S2P-, SARS-CoV spike protein-, a CoV spike protein-immobilized surfaces (blue, red, and green, respectively). ( C ) Amino acid sequences of three unique clones identified from panning (left) and their relative frequencies (%) (right). The sequences were aligned using the Kabat numbering system [ 38 ]. ELISA, enzyme-linked immunosorbent assay; MERS-S2P, Middle East respiratory syndrome-CoV S2 subunit protein; SARS-SP, severe acute respiratory syndrome-CoV S protein; HKU1-SP, hCoV HKU1 S protein; CoV, coronavirus; CDR, complementarity-determining region; FR, framework region.

    Journal: Antibodies

    Article Title: Selection and Characterization of Monoclonal Antibodies Targeting Middle East Respiratory Syndrome Coronavirus through a Human Synthetic Fab Phage Display Library Panning

    doi: 10.3390/antib8030042

    Figure Lengend Snippet: Output of the panning of the phage-displayed synthetic Fab library on MERS-S2P. ( A ) Monitoring of phage titers over three rounds (R1–R3) of panning. Black and gray bars indicate a ratio of phage output to input titers presented as a percentage (%) from panning on MERS-S2P-immobilized and -non-immobilized surfaces, respectively. The ratio of output to input (%) = (phage output titer ÷ phage input titer) × 100. ( B ) Phage ELISAs performed on MERS-S2P-, SARS-CoV spike protein-, a CoV spike protein-immobilized surfaces (blue, red, and green, respectively). ( C ) Amino acid sequences of three unique clones identified from panning (left) and their relative frequencies (%) (right). The sequences were aligned using the Kabat numbering system [ 38 ]. ELISA, enzyme-linked immunosorbent assay; MERS-S2P, Middle East respiratory syndrome-CoV S2 subunit protein; SARS-SP, severe acute respiratory syndrome-CoV S protein; HKU1-SP, hCoV HKU1 S protein; CoV, coronavirus; CDR, complementarity-determining region; FR, framework region.

    Article Snippet: The rabbit anti-MERS-CoV antibodies (1:3000, Sino Biological), serially two-fold-diluted MERS-S2P, and HRP-conjugated goat anti-rabbit antibodies (1:6000, Sigma Aldrich) were added and incubated for 30 min at RT.

    Techniques: Clone Assay, Enzyme-linked Immunosorbent Assay

    Detection of MERS-S2P using S2A3 (IgG) on ACCEL ELISA™ plates. ( A ) Schematic depicting the sandwich ELISA format to detect MERS-S2P using S2A3 (IgG) (capture antibody) and rabbit anti-MERS-CoV IgG (detection antibody) on ACCEL ELISA™ plates. ( B ) ELISA detection of MERS-S2P on a capture antibody (S2A3 (IgG)) immobilized using three different concentrations (3 µg/mL, 5 µg/mL, and 10 µg/mL) on ACCEL ELISA™ plates. The goodness of fit is indicated by the R 2 value. LOD, limit of detection.

    Journal: Antibodies

    Article Title: Selection and Characterization of Monoclonal Antibodies Targeting Middle East Respiratory Syndrome Coronavirus through a Human Synthetic Fab Phage Display Library Panning

    doi: 10.3390/antib8030042

    Figure Lengend Snippet: Detection of MERS-S2P using S2A3 (IgG) on ACCEL ELISA™ plates. ( A ) Schematic depicting the sandwich ELISA format to detect MERS-S2P using S2A3 (IgG) (capture antibody) and rabbit anti-MERS-CoV IgG (detection antibody) on ACCEL ELISA™ plates. ( B ) ELISA detection of MERS-S2P on a capture antibody (S2A3 (IgG)) immobilized using three different concentrations (3 µg/mL, 5 µg/mL, and 10 µg/mL) on ACCEL ELISA™ plates. The goodness of fit is indicated by the R 2 value. LOD, limit of detection.

    Article Snippet: The rabbit anti-MERS-CoV antibodies (1:3000, Sino Biological), serially two-fold-diluted MERS-S2P, and HRP-conjugated goat anti-rabbit antibodies (1:6000, Sigma Aldrich) were added and incubated for 30 min at RT.

    Techniques: Enzyme-linked Immunosorbent Assay, Sandwich ELISA

    Mapping of H2-d restricted T cell epitopes in MERS-CoV N protein; ( a–b ) BALB/c mice (n = 2 to 4) were immunized twice (21-day interval) i.p. or i.m. with 10 8 PFU of recombinant MVA-MERS-N (MVA-N), non-recombinant MVA (MVA) or PBS. Splenocytes from vaccinated mice were incubated in the presence of subpools (V8.1, V8.2, H8.1, H8.2) from positive matrix pools ( a ) or individual 15-mers peptides #89 or #90 ( b ). IFN-γ spot-forming CD8+ T cells (IFN-γ SFC) were quantified by ELISPOT. The lines represent means.

    Journal: Viruses

    Article Title: CD8+ T Cells Responding to the Middle East Respiratory Syndrome Coronavirus Nucleocapsid Protein Delivered by Vaccinia Virus MVA in Mice

    doi: 10.3390/v10120718

    Figure Lengend Snippet: Mapping of H2-d restricted T cell epitopes in MERS-CoV N protein; ( a–b ) BALB/c mice (n = 2 to 4) were immunized twice (21-day interval) i.p. or i.m. with 10 8 PFU of recombinant MVA-MERS-N (MVA-N), non-recombinant MVA (MVA) or PBS. Splenocytes from vaccinated mice were incubated in the presence of subpools (V8.1, V8.2, H8.1, H8.2) from positive matrix pools ( a ) or individual 15-mers peptides #89 or #90 ( b ). IFN-γ spot-forming CD8+ T cells (IFN-γ SFC) were quantified by ELISPOT. The lines represent means.

    Article Snippet: After 1 h blocking in a phosphate buffered saline (PBS) buffer containing 1% (w/v) non-fat dried milk and 0.1% (v/v) NP-40 detergent, the blots were incubated with monoclonal mouse anti-MERS-CoV Nucleocapsid antibody (Sino Biological, Beijing, China, 1:1000), monoclonal rabbit anti-MERS-CoV Spike Protein S1 Antibody (Sino Biological, 1:500), or polyclonal sera from MERS-CoV infected rabbits or cynomolgus macaques (kindly provided by Dr. Bart Haagmans, Erasmus Medical Center, Rotterdam, 1:1000) [ ] as primary antibodies.

    Techniques: Mouse Assay, Recombinant, Incubation, Enzyme-linked Immunospot

    Identification of an H2-d restricted T cell epitope in MERS-CoV N protein; ( a–d ) Groups of BALB/c mice ( n = 3 to 8) were vaccinated in a prime-boost regime with 10 8 PFU of MVA-MERS-N via i.p. ( a ) or i.m. ( b–d ) application. Mice immunized with non-recombinant MVA (MVA) and PBS served as negative controls. ( a-b ) Splenocytes were stimulated with individual 8-11-mer peptides and IFN-γ spot-forming CD8+ T cells (IFN-γ SFC) were measured by ELISPOT. ( c–d ) Splenocytes were stimulated with positive MERS-CoV N 10.2 peptide ( c ) or F2L 26-34 peptide ( d ) and IFN-γ producing CD8+ or CD4+ T cells were measured using intracellular cytokine staining assay and FACS analysis. The lines represent means. *

    Journal: Viruses

    Article Title: CD8+ T Cells Responding to the Middle East Respiratory Syndrome Coronavirus Nucleocapsid Protein Delivered by Vaccinia Virus MVA in Mice

    doi: 10.3390/v10120718

    Figure Lengend Snippet: Identification of an H2-d restricted T cell epitope in MERS-CoV N protein; ( a–d ) Groups of BALB/c mice ( n = 3 to 8) were vaccinated in a prime-boost regime with 10 8 PFU of MVA-MERS-N via i.p. ( a ) or i.m. ( b–d ) application. Mice immunized with non-recombinant MVA (MVA) and PBS served as negative controls. ( a-b ) Splenocytes were stimulated with individual 8-11-mer peptides and IFN-γ spot-forming CD8+ T cells (IFN-γ SFC) were measured by ELISPOT. ( c–d ) Splenocytes were stimulated with positive MERS-CoV N 10.2 peptide ( c ) or F2L 26-34 peptide ( d ) and IFN-γ producing CD8+ or CD4+ T cells were measured using intracellular cytokine staining assay and FACS analysis. The lines represent means. *

    Article Snippet: After 1 h blocking in a phosphate buffered saline (PBS) buffer containing 1% (w/v) non-fat dried milk and 0.1% (v/v) NP-40 detergent, the blots were incubated with monoclonal mouse anti-MERS-CoV Nucleocapsid antibody (Sino Biological, Beijing, China, 1:1000), monoclonal rabbit anti-MERS-CoV Spike Protein S1 Antibody (Sino Biological, 1:500), or polyclonal sera from MERS-CoV infected rabbits or cynomolgus macaques (kindly provided by Dr. Bart Haagmans, Erasmus Medical Center, Rotterdam, 1:1000) [ ] as primary antibodies.

    Techniques: Mouse Assay, Recombinant, Enzyme-linked Immunospot, Staining, FACS

    Analysis of recombinant MVA-MERS proteins; ( a ) Western Blot analysis of MERS-CoV N protein produced in CEF or HaCat cells. Lysates from cells infected with recombinant MVA (MVA-MERS-N, MVA-MERS-S) or non-recombinant MVA (MVA) at a MOI of five, or from non-infected cells (mock) were prepared at eight, 12, or 24 hpi. Proteins were analyzed by immunoblotting with a monoclonal anti-MERS-N antibody; ( b – d ) Western Blot analysis of MERS-CoV N and S proteins produced in CEF. Total cell extracts from CEF infected with recombinant MVA (MVA-MERS-N, MVA-MERS-S) or non-recombinant MVA (MVA) at a MOI of five, or from non-infected cells (mock) were prepared at 24 hpi. Cell lysates and proteins were tested by immunoblotting using monoclonal anti MERS-N and anti MERS-S antibody ( b ) or polyclonal sera from MERS-CoV infected rabbits ( c ) or cynomolgus macaques ( d ). Arrows indicate the N- or S-specific protein bands.

    Journal: Viruses

    Article Title: CD8+ T Cells Responding to the Middle East Respiratory Syndrome Coronavirus Nucleocapsid Protein Delivered by Vaccinia Virus MVA in Mice

    doi: 10.3390/v10120718

    Figure Lengend Snippet: Analysis of recombinant MVA-MERS proteins; ( a ) Western Blot analysis of MERS-CoV N protein produced in CEF or HaCat cells. Lysates from cells infected with recombinant MVA (MVA-MERS-N, MVA-MERS-S) or non-recombinant MVA (MVA) at a MOI of five, or from non-infected cells (mock) were prepared at eight, 12, or 24 hpi. Proteins were analyzed by immunoblotting with a monoclonal anti-MERS-N antibody; ( b – d ) Western Blot analysis of MERS-CoV N and S proteins produced in CEF. Total cell extracts from CEF infected with recombinant MVA (MVA-MERS-N, MVA-MERS-S) or non-recombinant MVA (MVA) at a MOI of five, or from non-infected cells (mock) were prepared at 24 hpi. Cell lysates and proteins were tested by immunoblotting using monoclonal anti MERS-N and anti MERS-S antibody ( b ) or polyclonal sera from MERS-CoV infected rabbits ( c ) or cynomolgus macaques ( d ). Arrows indicate the N- or S-specific protein bands.

    Article Snippet: After 1 h blocking in a phosphate buffered saline (PBS) buffer containing 1% (w/v) non-fat dried milk and 0.1% (v/v) NP-40 detergent, the blots were incubated with monoclonal mouse anti-MERS-CoV Nucleocapsid antibody (Sino Biological, Beijing, China, 1:1000), monoclonal rabbit anti-MERS-CoV Spike Protein S1 Antibody (Sino Biological, 1:500), or polyclonal sera from MERS-CoV infected rabbits or cynomolgus macaques (kindly provided by Dr. Bart Haagmans, Erasmus Medical Center, Rotterdam, 1:1000) [ ] as primary antibodies.

    Techniques: Recombinant, Western Blot, Produced, Infection

    Screening for H2-d restricted T cell epitopes in MERS-CoV N protein using matrix peptide pools; ( a – b ) groups of BALB/c mice ( n = 2 to 6) were vaccinated twice (21-day interval) by i.p. ( a ) or i.m. ( b ) application with 10 8 plaque-forming-units (PFU) of recombinant MVA-MERS-N (MVA-N). Mice inoculated with non-recombinant MVA (MVA) or phosphate-buffered saline (PBS) were used as controls. Splenocytes were restimulated in vitro with pools of overlapping peptides corresponding to MERS-CoV N protein. IFN-γ spot-forming CD8+ T cells (IFN-γ SFC) were measured by ELISPOT. The lines represent means.

    Journal: Viruses

    Article Title: CD8+ T Cells Responding to the Middle East Respiratory Syndrome Coronavirus Nucleocapsid Protein Delivered by Vaccinia Virus MVA in Mice

    doi: 10.3390/v10120718

    Figure Lengend Snippet: Screening for H2-d restricted T cell epitopes in MERS-CoV N protein using matrix peptide pools; ( a – b ) groups of BALB/c mice ( n = 2 to 6) were vaccinated twice (21-day interval) by i.p. ( a ) or i.m. ( b ) application with 10 8 plaque-forming-units (PFU) of recombinant MVA-MERS-N (MVA-N). Mice inoculated with non-recombinant MVA (MVA) or phosphate-buffered saline (PBS) were used as controls. Splenocytes were restimulated in vitro with pools of overlapping peptides corresponding to MERS-CoV N protein. IFN-γ spot-forming CD8+ T cells (IFN-γ SFC) were measured by ELISPOT. The lines represent means.

    Article Snippet: After 1 h blocking in a phosphate buffered saline (PBS) buffer containing 1% (w/v) non-fat dried milk and 0.1% (v/v) NP-40 detergent, the blots were incubated with monoclonal mouse anti-MERS-CoV Nucleocapsid antibody (Sino Biological, Beijing, China, 1:1000), monoclonal rabbit anti-MERS-CoV Spike Protein S1 Antibody (Sino Biological, 1:500), or polyclonal sera from MERS-CoV infected rabbits or cynomolgus macaques (kindly provided by Dr. Bart Haagmans, Erasmus Medical Center, Rotterdam, 1:1000) [ ] as primary antibodies.

    Techniques: Mouse Assay, Recombinant, In Vitro, Enzyme-linked Immunospot