mers  (Sino Biological)


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    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
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    Structured Review

    Sino Biological mers
    Identification of the binders by phage ELISA. (A) Monoclonal phage ELISA was carried out to identify the binders to GPC3, <t>Her2,</t> PD1, <t>MERS</t> 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.
    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 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    mers - by Bioz Stars, 2021-04
    94/100 stars

    Images

    1) 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 ( ).

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    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: 95/100, 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 95 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
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    95
    Sino Biological rabbit polyclonal antiserum against mers cov
    Sequencing of <t>MERS-CoV</t> Spike protein and data analysis
    Rabbit Polyclonal Antiserum Against Mers Cov, supplied by Sino Biological, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rabbit polyclonal antiserum against mers cov/product/Sino Biological
    Average 95 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
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    99
    Sino Biological mers cov spike protein
    Phylogenetic analysis of a partial spike S2 domain. A maximum likelihood tree based on the GTR+G+I model using 1000 bootstraps was generated from a spike S2 domain genome fragment corresponding to nucleotides 23781–24395 of HCoV-EMC/2012. The newly identified <t>MERS-CoV</t> sequences are depicted in bold , recent MERS-CoV sequences associated with an outbreak in Jordan in 2015 are depicted in bold . Bootstrap values of
    Mers Cov Spike Protein, supplied by Sino Biological, used in various techniques. Bioz Stars score: 99/100, 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
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    Image Search Results


    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

    Sequencing of MERS-CoV Spike protein and data analysis

    Journal: Virology

    Article Title: Intratracheal exposure of common marmosets to MERS-CoV Jordan-n3/2012 or MERS-CoV EMC/2012 isolates does not result in lethal disease

    doi: 10.1016/j.virol.2015.07.013

    Figure Lengend Snippet: Sequencing of MERS-CoV Spike protein and data analysis

    Article Snippet: To detect MERS-CoV antigen, IHC was performed using a rabbit polyclonal antiserum against MERS-CoV (Sino Biological PR China) (1:1000).

    Techniques: Sequencing

    Gross and Histopathology. (A) Gross lung pathology demonstrating mostly normal lung with multifocal to coalescing moderate interstitial pneumonia in the left caudal lung lobe. (B) Low magnification of lung field from MERS-CoV EMC inoculated subject demonstrating

    Journal: Virology

    Article Title: Intratracheal exposure of common marmosets to MERS-CoV Jordan-n3/2012 or MERS-CoV EMC/2012 isolates does not result in lethal disease

    doi: 10.1016/j.virol.2015.07.013

    Figure Lengend Snippet: Gross and Histopathology. (A) Gross lung pathology demonstrating mostly normal lung with multifocal to coalescing moderate interstitial pneumonia in the left caudal lung lobe. (B) Low magnification of lung field from MERS-CoV EMC inoculated subject demonstrating

    Article Snippet: To detect MERS-CoV antigen, IHC was performed using a rabbit polyclonal antiserum against MERS-CoV (Sino Biological PR China) (1:1000).

    Techniques: Histopathology

    Ex-vivo analysis of primary cells. Cells were isolated from lung, kidney, and bronchoalveolar lavage (BAL), and one-step growth kinetics were performed as described in Materials and Methods. Lung, Kidney and BAL demonstrate that MERS-CoV is able to replicate

    Journal: Virology

    Article Title: Intratracheal exposure of common marmosets to MERS-CoV Jordan-n3/2012 or MERS-CoV EMC/2012 isolates does not result in lethal disease

    doi: 10.1016/j.virol.2015.07.013

    Figure Lengend Snippet: Ex-vivo analysis of primary cells. Cells were isolated from lung, kidney, and bronchoalveolar lavage (BAL), and one-step growth kinetics were performed as described in Materials and Methods. Lung, Kidney and BAL demonstrate that MERS-CoV is able to replicate

    Article Snippet: To detect MERS-CoV antigen, IHC was performed using a rabbit polyclonal antiserum against MERS-CoV (Sino Biological PR China) (1:1000).

    Techniques: Ex Vivo, Isolation

    Fluorescence Reduction Neutralizing Assay. Sera from all subjects was evaluated for the presence of neutralizing antibody to MERS-CoV as described in Materials and Methods. Neutralizing antibody could be detected above the background of the Mock infected

    Journal: Virology

    Article Title: Intratracheal exposure of common marmosets to MERS-CoV Jordan-n3/2012 or MERS-CoV EMC/2012 isolates does not result in lethal disease

    doi: 10.1016/j.virol.2015.07.013

    Figure Lengend Snippet: Fluorescence Reduction Neutralizing Assay. Sera from all subjects was evaluated for the presence of neutralizing antibody to MERS-CoV as described in Materials and Methods. Neutralizing antibody could be detected above the background of the Mock infected

    Article Snippet: To detect MERS-CoV antigen, IHC was performed using a rabbit polyclonal antiserum against MERS-CoV (Sino Biological PR China) (1:1000).

    Techniques: Fluorescence, Neutralizing Assay, Infection

    Two Selected G2-escape Mutations and Their Impact on G2 Binding and Neutralization. (A-D) Binding of G2 Fab to immobilized (A) MERS-CoV S1-NTD, (B) MERS-CoV S-2P, (C) MERS-CoV S-2P-S28F and (D) MERS-CoV S-2P-G198D measured by surface plasmon resonance (SPR). The same concentration series of G2 Fab was used in A-D. Best global fit of the data to a 1:1 binding model is shown as colored lines. (E) Neutralization activity of G2 IgG was measured against pseudotyped lentivirus bearing MERS-CoV S (WT) and two variants (S28F and G198D). Percent neutralization of WT (red), S28F (blue) and G198D (black) S pseudovirions at the different antibody concentrations is shown. Data points represent the mean of three technical replicates with standard errors. (F) Neutralization activity of G2 IgG was measured against authentic MERS-CoV (WT) and the G198D variant. Percent neutralization of WT (red) and G198D (black) MERS-CoV at the different antibody concentrations is shown. Data points for the wild-type virus represent the mean of two technical replicates.

    Journal: Cell reports

    Article Title: Structural Definition of a Neutralization-sensitive Epitope on the MERS-CoV S1-NTD

    doi: 10.1016/j.celrep.2019.08.052

    Figure Lengend Snippet: Two Selected G2-escape Mutations and Their Impact on G2 Binding and Neutralization. (A-D) Binding of G2 Fab to immobilized (A) MERS-CoV S1-NTD, (B) MERS-CoV S-2P, (C) MERS-CoV S-2P-S28F and (D) MERS-CoV S-2P-G198D measured by surface plasmon resonance (SPR). The same concentration series of G2 Fab was used in A-D. Best global fit of the data to a 1:1 binding model is shown as colored lines. (E) Neutralization activity of G2 IgG was measured against pseudotyped lentivirus bearing MERS-CoV S (WT) and two variants (S28F and G198D). Percent neutralization of WT (red), S28F (blue) and G198D (black) S pseudovirions at the different antibody concentrations is shown. Data points represent the mean of three technical replicates with standard errors. (F) Neutralization activity of G2 IgG was measured against authentic MERS-CoV (WT) and the G198D variant. Percent neutralization of WT (red) and G198D (black) MERS-CoV at the different antibody concentrations is shown. Data points for the wild-type virus represent the mean of two technical replicates.

    Article Snippet: Subsequently, cells were stained with MERS-CoV S rabbit polyclonal antibodies (Sino Biological, Beijing, China) and then secondary goat anti-rabbit IgG H & L labeled with Alexa Fluor® 488 (AF488) was added.

    Techniques: Binding Assay, Neutralization, SPR Assay, Concentration Assay, Activity Assay, Variant Assay

    G2 IgG Prevents the Binding of MERS-CoV S Protein to DDP4-Expressing Cells. Normalized binding efficiency of GFP-tagged MERS-CoV S-2P proteins to DPP4-expressing FreeStyle 293F cells in the presence or absence of IgGs was calculated from median fluorescence intensity (MFI) values. FreeStyle 293-F cells were transfected with a plasmid encoding full-length DPP4 60 h before the experiment. Non-transfected cells (NT) incubated with MERS-CoV S-2P, as well as transfected cells incubated with PBS, were used as negative controls. AM14 is an irrelevant RSV F-specific neutralizing antibody used as another negative control. Bar graph shows the mean and error bars indicate the standard deviation (n = 3 biologically independent experiments with two technical replicates).

    Journal: Cell reports

    Article Title: Structural Definition of a Neutralization-sensitive Epitope on the MERS-CoV S1-NTD

    doi: 10.1016/j.celrep.2019.08.052

    Figure Lengend Snippet: G2 IgG Prevents the Binding of MERS-CoV S Protein to DDP4-Expressing Cells. Normalized binding efficiency of GFP-tagged MERS-CoV S-2P proteins to DPP4-expressing FreeStyle 293F cells in the presence or absence of IgGs was calculated from median fluorescence intensity (MFI) values. FreeStyle 293-F cells were transfected with a plasmid encoding full-length DPP4 60 h before the experiment. Non-transfected cells (NT) incubated with MERS-CoV S-2P, as well as transfected cells incubated with PBS, were used as negative controls. AM14 is an irrelevant RSV F-specific neutralizing antibody used as another negative control. Bar graph shows the mean and error bars indicate the standard deviation (n = 3 biologically independent experiments with two technical replicates).

    Article Snippet: Subsequently, cells were stained with MERS-CoV S rabbit polyclonal antibodies (Sino Biological, Beijing, China) and then secondary goat anti-rabbit IgG H & L labeled with Alexa Fluor® 488 (AF488) was added.

    Techniques: Binding Assay, Expressing, Fluorescence, Transfection, Plasmid Preparation, Incubation, Negative Control, Standard Deviation

    G2-mediated Inhibition of DPP4 binding to MERS-CoV S Depends on Oligomeric State of the Spike (A–C) Normalized binding efficiency of DPP4, in the presence or absence of Fab, to cells transfected with plasmids encoding (A) membrane-anchored S1 (S1-TM), (B) full-length S-WT (S-WT-FL) and (C) full-length S-2P (S-2P-FL). Cells incubated with PBS were used as negative controls. Bar graph shows the mean and error bars indicate the standard deviation (n = 3 biologically independent experiments with two technical replicates). (D) Surface plasmon resonance competition assay. Response curves for MERS-CoV S-2P, alone or in the presence of a 5-fold molar excess of indicated Fabs or IgGs, passed over immobilized DPP4 ectodomain. The curves for S-2P or S-2P supplemented with Fab or IgG are shown with a solid line, whereas control curves for samples without S-2P are shown with a dotted line.

    Journal: Cell reports

    Article Title: Structural Definition of a Neutralization-sensitive Epitope on the MERS-CoV S1-NTD

    doi: 10.1016/j.celrep.2019.08.052

    Figure Lengend Snippet: G2-mediated Inhibition of DPP4 binding to MERS-CoV S Depends on Oligomeric State of the Spike (A–C) Normalized binding efficiency of DPP4, in the presence or absence of Fab, to cells transfected with plasmids encoding (A) membrane-anchored S1 (S1-TM), (B) full-length S-WT (S-WT-FL) and (C) full-length S-2P (S-2P-FL). Cells incubated with PBS were used as negative controls. Bar graph shows the mean and error bars indicate the standard deviation (n = 3 biologically independent experiments with two technical replicates). (D) Surface plasmon resonance competition assay. Response curves for MERS-CoV S-2P, alone or in the presence of a 5-fold molar excess of indicated Fabs or IgGs, passed over immobilized DPP4 ectodomain. The curves for S-2P or S-2P supplemented with Fab or IgG are shown with a solid line, whereas control curves for samples without S-2P are shown with a dotted line.

    Article Snippet: Subsequently, cells were stained with MERS-CoV S rabbit polyclonal antibodies (Sino Biological, Beijing, China) and then secondary goat anti-rabbit IgG H & L labeled with Alexa Fluor® 488 (AF488) was added.

    Techniques: Inhibition, Binding Assay, Transfection, Incubation, Standard Deviation, SPR Assay, Competitive Binding Assay

    Phylogenetic analysis of a partial spike S2 domain. A maximum likelihood tree based on the GTR+G+I model using 1000 bootstraps was generated from a spike S2 domain genome fragment corresponding to nucleotides 23781–24395 of HCoV-EMC/2012. The newly identified MERS-CoV sequences are depicted in bold , recent MERS-CoV sequences associated with an outbreak in Jordan in 2015 are depicted in bold . Bootstrap values of

    Journal: Vector Borne and Zoonotic Diseases

    Article Title: High Prevalence of Middle East Respiratory Coronavirus in Young Dromedary Camels in Jordan

    doi: 10.1089/vbz.2016.2062

    Figure Lengend Snippet: Phylogenetic analysis of a partial spike S2 domain. A maximum likelihood tree based on the GTR+G+I model using 1000 bootstraps was generated from a spike S2 domain genome fragment corresponding to nucleotides 23781–24395 of HCoV-EMC/2012. The newly identified MERS-CoV sequences are depicted in bold , recent MERS-CoV sequences associated with an outbreak in Jordan in 2015 are depicted in bold . Bootstrap values of

    Article Snippet: 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.

    Techniques: Generated