rabbit anti mers cov antibodies Search Results


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    Sino Biological mers cov nucleocapsid protein antibody rabbit pab
    <t>MERS-CoV</t> loads in the swabs and tissues of immunised rhesus macaques following MERS-CoV infection detected by Real-time-PCR. a. Viral loads in swab samples at 1 dpi and 3 dpi. b. The viral loads in lung and trachea tissue 3 days after MERS-CoV infection. All of the detections were replicated for three times. M, H and L represent mock, high- and low-dose groups, respectively. * p
    Mers Cov Nucleocapsid Protein Antibody Rabbit Pab, 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
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    94
    Sino Biological rabbit anti mers cov spike antibody
    CEACAM5 overexpression does not confer infectivity to nonpermissive cells on <t>MERS-CoV</t> but enhances MERS-CoV entry into permissive cells. (A and C) To verify the role of CEACAM5 in MERS-CoV entry, human CEACAM5 was overexpressed in BHK21 cells or AD293 cells. The cells were then challenged with MERS-CoV at an MOI of 1 for 2 h at 37°C. After 2 h, the cells were washed extensively and harvested for qPCR analysis. (B and D) To verify the role of CEACAM5 in MERS-CoV attachment, human CEACAM5-expressing BHK21 cells or human CEACAM5-expressing AD293 cells were challenged with MERS-CoV at an MOI of 1 for 2 h at 4°C. The cell lysates were then harvested for qPCR analysis. DPP4- and empty-vector (pcDNA3.1)-expressing BHK21 cells were included as controls. The data are represented as means and standard deviations of the results of three independent experiments. The results from the human DPP4-, human CEACAM5-, and empty-vector-overexpressing samples were compared with those of the mock-treated samples. Statistical analyses were carried out using Student's t test. Statistical significance is indicated by the asterisks ( P
    Rabbit Anti Mers Cov Spike Antibody, supplied by Sino Biological, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rabbit anti mers cov spike antibody/product/Sino Biological
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    rabbit anti mers cov spike antibody - by Bioz Stars, 2021-06
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    94
    Sino Biological rabbit monoclonal antibodies
    CEACAM5 overexpression does not confer infectivity to nonpermissive cells on <t>MERS-CoV</t> but enhances MERS-CoV entry into permissive cells. (A and C) To verify the role of CEACAM5 in MERS-CoV entry, human CEACAM5 was overexpressed in BHK21 cells or AD293 cells. The cells were then challenged with MERS-CoV at an MOI of 1 for 2 h at 37°C. After 2 h, the cells were washed extensively and harvested for qPCR analysis. (B and D) To verify the role of CEACAM5 in MERS-CoV attachment, human CEACAM5-expressing BHK21 cells or human CEACAM5-expressing AD293 cells were challenged with MERS-CoV at an MOI of 1 for 2 h at 4°C. The cell lysates were then harvested for qPCR analysis. DPP4- and empty-vector (pcDNA3.1)-expressing BHK21 cells were included as controls. The data are represented as means and standard deviations of the results of three independent experiments. The results from the human DPP4-, human CEACAM5-, and empty-vector-overexpressing samples were compared with those of the mock-treated samples. Statistical analyses were carried out using Student's t test. Statistical significance is indicated by the asterisks ( P
    Rabbit Monoclonal Antibodies, supplied by Sino Biological, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rabbit monoclonal antibodies/product/Sino Biological
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    94
    Sino Biological rabbit polyclonal anti mers cov spike protein antibody
    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 Polyclonal Anti Mers Cov Spike Protein Antibody, supplied by Sino Biological, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rabbit polyclonal anti mers cov spike protein antibody/product/Sino Biological
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    rabbit polyclonal anti mers cov spike protein antibody - by Bioz Stars, 2021-06
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    Image Search Results


    MERS-CoV loads in the swabs and tissues of immunised rhesus macaques following MERS-CoV infection detected by Real-time-PCR. a. Viral loads in swab samples at 1 dpi and 3 dpi. b. The viral loads in lung and trachea tissue 3 days after MERS-CoV infection. All of the detections were replicated for three times. M, H and L represent mock, high- and low-dose groups, respectively. * p

    Journal: EBioMedicine

    Article Title: Recombinant Receptor Binding Domain Protein Induces Partial Protective Immunity in Rhesus Macaques Against Middle East Respiratory Syndrome Coronavirus Challenge

    doi: 10.1016/j.ebiom.2015.08.031

    Figure Lengend Snippet: MERS-CoV loads in the swabs and tissues of immunised rhesus macaques following MERS-CoV infection detected by Real-time-PCR. a. Viral loads in swab samples at 1 dpi and 3 dpi. b. The viral loads in lung and trachea tissue 3 days after MERS-CoV infection. All of the detections were replicated for three times. M, H and L represent mock, high- and low-dose groups, respectively. * p

    Article Snippet: A rabbit-serum-derived polyclonal antibody against nucleoprotein (Sino Biological Inc., cat: 100213-RP02) at 1:1000 dilution, was then incubated with the sections.

    Techniques: Infection, Real-time Polymerase Chain Reaction

    IHC staining of immunised rhesus macaque lung and trachea 3 days after MERS-CoV infection by a polyclonal antibody against nucleoprotein. a–c. IHC analysis of lung tissue. d–f. IHC staining of trachea tissue. Black arrows indicate the distribution of viral antigen. M, H and L represent mock, high- and low-dose groups, respectively.

    Journal: EBioMedicine

    Article Title: Recombinant Receptor Binding Domain Protein Induces Partial Protective Immunity in Rhesus Macaques Against Middle East Respiratory Syndrome Coronavirus Challenge

    doi: 10.1016/j.ebiom.2015.08.031

    Figure Lengend Snippet: IHC staining of immunised rhesus macaque lung and trachea 3 days after MERS-CoV infection by a polyclonal antibody against nucleoprotein. a–c. IHC analysis of lung tissue. d–f. IHC staining of trachea tissue. Black arrows indicate the distribution of viral antigen. M, H and L represent mock, high- and low-dose groups, respectively.

    Article Snippet: A rabbit-serum-derived polyclonal antibody against nucleoprotein (Sino Biological Inc., cat: 100213-RP02) at 1:1000 dilution, was then incubated with the sections.

    Techniques: Immunohistochemistry, Staining, Infection

    Schematic diagram of the rRBD vaccination and MERS-CoV challenge schedule. Monkeys were immunised three times intramuscularly (i.m.), at intervals of 8 or 17 weeks, and inoculated intratracheally with hCoV-EMC at a dosage of 6.5 × 10 7 TCID 50 (red arrow). Monkeys were bled periodically at 2, 10, and 25 weeks (− 14 days), and at − 1 day, 1 day and 3 days. Additionally, nasal, oral and rectal swabs were collected at − 1 day, 1 day and 3 days. Chest X-rays were performed 1 day before inoculation, and at 1 and 3 dpi. Following euthanasia, lung, trachea, spleen and kidney specimens of challenged monkeys were acquired at 3 dpi for detection.

    Journal: EBioMedicine

    Article Title: Recombinant Receptor Binding Domain Protein Induces Partial Protective Immunity in Rhesus Macaques Against Middle East Respiratory Syndrome Coronavirus Challenge

    doi: 10.1016/j.ebiom.2015.08.031

    Figure Lengend Snippet: Schematic diagram of the rRBD vaccination and MERS-CoV challenge schedule. Monkeys were immunised three times intramuscularly (i.m.), at intervals of 8 or 17 weeks, and inoculated intratracheally with hCoV-EMC at a dosage of 6.5 × 10 7 TCID 50 (red arrow). Monkeys were bled periodically at 2, 10, and 25 weeks (− 14 days), and at − 1 day, 1 day and 3 days. Additionally, nasal, oral and rectal swabs were collected at − 1 day, 1 day and 3 days. Chest X-rays were performed 1 day before inoculation, and at 1 and 3 dpi. Following euthanasia, lung, trachea, spleen and kidney specimens of challenged monkeys were acquired at 3 dpi for detection.

    Article Snippet: A rabbit-serum-derived polyclonal antibody against nucleoprotein (Sino Biological Inc., cat: 100213-RP02) at 1:1000 dilution, was then incubated with the sections.

    Techniques:

    Radiographic alterations and lung pathology. a. X-rays from rhesus macaques imaged prior to (− 1 day) and post-MERS-CoV inoculation (1 day and 3 days). Areas of interstitial infiltration, indicative of pneumonia, are highlighted (circle). b. Gross pathology of the lungs from necropsied animals at 3 dpi. Haemorrhage and necrosis indicated by the black circle in both high and low dose immunisation groups were small and local, which happened in the mock group were large and disperse. M, H and L represent the mock, high- and low-dose groups, respectively.

    Journal: EBioMedicine

    Article Title: Recombinant Receptor Binding Domain Protein Induces Partial Protective Immunity in Rhesus Macaques Against Middle East Respiratory Syndrome Coronavirus Challenge

    doi: 10.1016/j.ebiom.2015.08.031

    Figure Lengend Snippet: Radiographic alterations and lung pathology. a. X-rays from rhesus macaques imaged prior to (− 1 day) and post-MERS-CoV inoculation (1 day and 3 days). Areas of interstitial infiltration, indicative of pneumonia, are highlighted (circle). b. Gross pathology of the lungs from necropsied animals at 3 dpi. Haemorrhage and necrosis indicated by the black circle in both high and low dose immunisation groups were small and local, which happened in the mock group were large and disperse. M, H and L represent the mock, high- and low-dose groups, respectively.

    Article Snippet: A rabbit-serum-derived polyclonal antibody against nucleoprotein (Sino Biological Inc., cat: 100213-RP02) at 1:1000 dilution, was then incubated with the sections.

    Techniques:

    Histopathological changes in the lungs and tracheas of immunised rhesus macaques following MERS-CoV challenge. Three days after MERS-CoV inoculation, all monkeys were euthanised. Tissues were collected and stained with haematoxylin and eosin (H E). a–f. Pathological changes in the lungs of immunised monkeys. The black triangle indicates acute interstitial pneumonia diffused in lung tissue. The unfilled arrow indicates the hyaline member resulting from effusion of fibrin. The black arrow indicates the influx of inflammatory cells. g–i. Pathological findings of tracheas in immunised rhesus macaques. The black arrow highlights the infiltration of inflammatory cells in the tunica mucosa bronchiorum. The unfilled arrow indicates epithelium impairment in tunica mucosa bronchiorum. M, H and L represent the mock, high- and low-dose groups, respectively.

    Journal: EBioMedicine

    Article Title: Recombinant Receptor Binding Domain Protein Induces Partial Protective Immunity in Rhesus Macaques Against Middle East Respiratory Syndrome Coronavirus Challenge

    doi: 10.1016/j.ebiom.2015.08.031

    Figure Lengend Snippet: Histopathological changes in the lungs and tracheas of immunised rhesus macaques following MERS-CoV challenge. Three days after MERS-CoV inoculation, all monkeys were euthanised. Tissues were collected and stained with haematoxylin and eosin (H E). a–f. Pathological changes in the lungs of immunised monkeys. The black triangle indicates acute interstitial pneumonia diffused in lung tissue. The unfilled arrow indicates the hyaline member resulting from effusion of fibrin. The black arrow indicates the influx of inflammatory cells. g–i. Pathological findings of tracheas in immunised rhesus macaques. The black arrow highlights the infiltration of inflammatory cells in the tunica mucosa bronchiorum. The unfilled arrow indicates epithelium impairment in tunica mucosa bronchiorum. M, H and L represent the mock, high- and low-dose groups, respectively.

    Article Snippet: A rabbit-serum-derived polyclonal antibody against nucleoprotein (Sino Biological Inc., cat: 100213-RP02) at 1:1000 dilution, was then incubated with the sections.

    Techniques: Staining

    CEACAM5 overexpression does not confer infectivity to nonpermissive cells on MERS-CoV but enhances MERS-CoV entry into permissive cells. (A and C) To verify the role of CEACAM5 in MERS-CoV entry, human CEACAM5 was overexpressed in BHK21 cells or AD293 cells. The cells were then challenged with MERS-CoV at an MOI of 1 for 2 h at 37°C. After 2 h, the cells were washed extensively and harvested for qPCR analysis. (B and D) To verify the role of CEACAM5 in MERS-CoV attachment, human CEACAM5-expressing BHK21 cells or human CEACAM5-expressing AD293 cells were challenged with MERS-CoV at an MOI of 1 for 2 h at 4°C. The cell lysates were then harvested for qPCR analysis. DPP4- and empty-vector (pcDNA3.1)-expressing BHK21 cells were included as controls. The data are represented as means and standard deviations of the results of three independent experiments. The results from the human DPP4-, human CEACAM5-, and empty-vector-overexpressing samples were compared with those of the mock-treated samples. Statistical analyses were carried out using Student's t test. Statistical significance is indicated by the asterisks ( P

    Journal: Journal of Virology

    Article Title: Carcinoembryonic Antigen-Related Cell Adhesion Molecule 5 Is an Important Surface Attachment Factor That Facilitates Entry of Middle East Respiratory Syndrome Coronavirus

    doi: 10.1128/JVI.01133-16

    Figure Lengend Snippet: CEACAM5 overexpression does not confer infectivity to nonpermissive cells on MERS-CoV but enhances MERS-CoV entry into permissive cells. (A and C) To verify the role of CEACAM5 in MERS-CoV entry, human CEACAM5 was overexpressed in BHK21 cells or AD293 cells. The cells were then challenged with MERS-CoV at an MOI of 1 for 2 h at 37°C. After 2 h, the cells were washed extensively and harvested for qPCR analysis. (B and D) To verify the role of CEACAM5 in MERS-CoV attachment, human CEACAM5-expressing BHK21 cells or human CEACAM5-expressing AD293 cells were challenged with MERS-CoV at an MOI of 1 for 2 h at 4°C. The cell lysates were then harvested for qPCR analysis. DPP4- and empty-vector (pcDNA3.1)-expressing BHK21 cells were included as controls. The data are represented as means and standard deviations of the results of three independent experiments. The results from the human DPP4-, human CEACAM5-, and empty-vector-overexpressing samples were compared with those of the mock-treated samples. Statistical analyses were carried out using Student's t test. Statistical significance is indicated by the asterisks ( P

    Article Snippet: MERS-CoV spike was detected either with the in-house mouse anti-MERS-CoV spike immune serum or with a rabbit anti-MERS-CoV spike antibody (Sino Biological, China).

    Techniques: Over Expression, Infection, Real-time Polymerase Chain Reaction, Expressing, Plasmid Preparation

    CEACAM5 recombinant protein and siRNA knockdown of CEACAM5 inhibit MERS-CoV entry. (A and B) MERS-CoV at an MOI of 0.1 was preincubated with human recombinant proteins at the indicated concentrations for 1 h at 37°C. After the preincubation, the protein-virus mixture was added to Calu3 cells (A) or Huh7 cells (B) for 2 h at 37°C. The cell lysates were subsequently harvested for qPCR analysis. Human recombinant DPP4 and human recombinant IgG were included as positive and negative controls, respectively. Results from the human recombinant DPP4-, human recombinant CEACAM5-, and control human recombinant IgG-treated samples were compared with those of the mock-treated samples. (C) Huh7 cells were treated with CEACAM5 protein for a total of 3 h during preincubation and virus inoculation (−1 to 2) or after virus inoculation (2 to 5). MERS-CoV entry was assessed by qPCR. (D) Huh7 cells were treated with 100 nM gene-specific or scrambled siRNA for two consecutive days. (E and F) Summary of the reduction of surface CEACAM5 and DPP4 expression. (G) siRNA-treated Huh7 cells were subjected to MERS-CoV infection at an MOI of 1 for 2 h at 37°C. The cell lysates were subsequently harvested for qPCR analysis, and the result was normalized to those of the mock-treated cells. The results from the DPP4 siRNA-, CEACAM5 siRNA-, and scrambled-siRNA-treated samples were compared with those of the mock-treated samples. The data are represented as means and standard deviations of the results of three independent experiments. Statistical analyses were carried out using Student's t test. Statistical significance is indicated by the asterisks ( P

    Journal: Journal of Virology

    Article Title: Carcinoembryonic Antigen-Related Cell Adhesion Molecule 5 Is an Important Surface Attachment Factor That Facilitates Entry of Middle East Respiratory Syndrome Coronavirus

    doi: 10.1128/JVI.01133-16

    Figure Lengend Snippet: CEACAM5 recombinant protein and siRNA knockdown of CEACAM5 inhibit MERS-CoV entry. (A and B) MERS-CoV at an MOI of 0.1 was preincubated with human recombinant proteins at the indicated concentrations for 1 h at 37°C. After the preincubation, the protein-virus mixture was added to Calu3 cells (A) or Huh7 cells (B) for 2 h at 37°C. The cell lysates were subsequently harvested for qPCR analysis. Human recombinant DPP4 and human recombinant IgG were included as positive and negative controls, respectively. Results from the human recombinant DPP4-, human recombinant CEACAM5-, and control human recombinant IgG-treated samples were compared with those of the mock-treated samples. (C) Huh7 cells were treated with CEACAM5 protein for a total of 3 h during preincubation and virus inoculation (−1 to 2) or after virus inoculation (2 to 5). MERS-CoV entry was assessed by qPCR. (D) Huh7 cells were treated with 100 nM gene-specific or scrambled siRNA for two consecutive days. (E and F) Summary of the reduction of surface CEACAM5 and DPP4 expression. (G) siRNA-treated Huh7 cells were subjected to MERS-CoV infection at an MOI of 1 for 2 h at 37°C. The cell lysates were subsequently harvested for qPCR analysis, and the result was normalized to those of the mock-treated cells. The results from the DPP4 siRNA-, CEACAM5 siRNA-, and scrambled-siRNA-treated samples were compared with those of the mock-treated samples. The data are represented as means and standard deviations of the results of three independent experiments. Statistical analyses were carried out using Student's t test. Statistical significance is indicated by the asterisks ( P

    Article Snippet: MERS-CoV spike was detected either with the in-house mouse anti-MERS-CoV spike immune serum or with a rabbit anti-MERS-CoV spike antibody (Sino Biological, China).

    Techniques: Recombinant, Real-time Polymerase Chain Reaction, Expressing, Infection

    CEACAM 5 interacts with MERS-CoV spike. (A) Surface and intracellular expression of CEACAM5-V5 were verified by flow cytometry. (B) For co-IP, BHK21 cells were transfected with CEACAM5-V5 (lanes 1 and 2 from left) or empty vector (lane 3). The cell lysate was immunoprecipitated with MERS-CoV S1-FLAG (lanes 1 and 3) or E. coli BAP-FLAG protein (lane 2) preadsorbed onto anti-FLAG M2 agarose beads prior to SDS-PAGE. The protein complex was detected by using the anti-FLAG antibody or the anti-V5 antibody. (C) Reciprocal co-IP was performed using CEACAM5-V5 as the bait. Purified MERS-CoV S1-FLAG (lanes 1 and 3) or BAP-FLAG (lane 2) proteins were immunoprecipitated with overexpressed CEACAM5-V5 or pcDNA-V5 protein preadsorbed onto anti-V5 Sepharose beads. Western blots were detected with the anti-FLAG or the anti-CEACAM5 antibody. (D) Endogenous co-IP was performed with MERS-CoV-infected or mock-infected Huh7 cell lysates using the rabbit anti-CEACAM5 antibody, the rabbit anti-MERS-CoV spike antibody, or the rabbit isotype IgG. Western blots were detected with the rabbit anti-MERS-CoV spike antibody or the rabbit anti-CEACAM5 antibody.

    Journal: Journal of Virology

    Article Title: Carcinoembryonic Antigen-Related Cell Adhesion Molecule 5 Is an Important Surface Attachment Factor That Facilitates Entry of Middle East Respiratory Syndrome Coronavirus

    doi: 10.1128/JVI.01133-16

    Figure Lengend Snippet: CEACAM 5 interacts with MERS-CoV spike. (A) Surface and intracellular expression of CEACAM5-V5 were verified by flow cytometry. (B) For co-IP, BHK21 cells were transfected with CEACAM5-V5 (lanes 1 and 2 from left) or empty vector (lane 3). The cell lysate was immunoprecipitated with MERS-CoV S1-FLAG (lanes 1 and 3) or E. coli BAP-FLAG protein (lane 2) preadsorbed onto anti-FLAG M2 agarose beads prior to SDS-PAGE. The protein complex was detected by using the anti-FLAG antibody or the anti-V5 antibody. (C) Reciprocal co-IP was performed using CEACAM5-V5 as the bait. Purified MERS-CoV S1-FLAG (lanes 1 and 3) or BAP-FLAG (lane 2) proteins were immunoprecipitated with overexpressed CEACAM5-V5 or pcDNA-V5 protein preadsorbed onto anti-V5 Sepharose beads. Western blots were detected with the anti-FLAG or the anti-CEACAM5 antibody. (D) Endogenous co-IP was performed with MERS-CoV-infected or mock-infected Huh7 cell lysates using the rabbit anti-CEACAM5 antibody, the rabbit anti-MERS-CoV spike antibody, or the rabbit isotype IgG. Western blots were detected with the rabbit anti-MERS-CoV spike antibody or the rabbit anti-CEACAM5 antibody.

    Article Snippet: MERS-CoV spike was detected either with the in-house mouse anti-MERS-CoV spike immune serum or with a rabbit anti-MERS-CoV spike antibody (Sino Biological, China).

    Techniques: Expressing, Flow Cytometry, Cytometry, Co-Immunoprecipitation Assay, Transfection, Plasmid Preparation, Immunoprecipitation, SDS Page, Purification, Western Blot, Infection

    CEACAM5 is a cell surface binding protein of MERS-CoV. (A) A549 and NIH 3T3 biotinylated cell membrane proteins were extracted, separated by 4 to 12% gradient gel, and transferred to PVDF membranes. WB, Western blotting. (B) Membrane proteins were probed with MERS-S-pseudotyped virus followed by detection of virus binding by incubation with immune serum directed against the MERS-CoV spike protein (left lane). Nonsusceptible cell membrane extracts from NIH 3T3 was included as a negative control (right lane). (C) The gel fractions corresponding to the positive signal at approximately 90 kDa were cut, electroeluted, and confirmed by VOPBA prior to submission for MS protein identification. (B and C) The position of CEACAM5 is indicated with asterisks. (D) Identified amino acid sequences and their corresponding positions.

    Journal: Journal of Virology

    Article Title: Carcinoembryonic Antigen-Related Cell Adhesion Molecule 5 Is an Important Surface Attachment Factor That Facilitates Entry of Middle East Respiratory Syndrome Coronavirus

    doi: 10.1128/JVI.01133-16

    Figure Lengend Snippet: CEACAM5 is a cell surface binding protein of MERS-CoV. (A) A549 and NIH 3T3 biotinylated cell membrane proteins were extracted, separated by 4 to 12% gradient gel, and transferred to PVDF membranes. WB, Western blotting. (B) Membrane proteins were probed with MERS-S-pseudotyped virus followed by detection of virus binding by incubation with immune serum directed against the MERS-CoV spike protein (left lane). Nonsusceptible cell membrane extracts from NIH 3T3 was included as a negative control (right lane). (C) The gel fractions corresponding to the positive signal at approximately 90 kDa were cut, electroeluted, and confirmed by VOPBA prior to submission for MS protein identification. (B and C) The position of CEACAM5 is indicated with asterisks. (D) Identified amino acid sequences and their corresponding positions.

    Article Snippet: MERS-CoV spike was detected either with the in-house mouse anti-MERS-CoV spike immune serum or with a rabbit anti-MERS-CoV spike antibody (Sino Biological, China).

    Techniques: Binding Assay, Western Blot, Incubation, Negative Control, Mass Spectrometry

    CEACAM5 is an attachment factor for MERS-CoV. (A and B) To assess whether CEACAM5 is important for MERS-CoV entry, human CEACAM5 was overexpressed in BHK21 cells. Human CEACAM5-expressing BHK21 cells were then challenged with MERS-CoV at an MOI of 5 for 2 h at 37°C. The inoculum was replaced with culture medium, and the cells were incubated for another 4 h before being harvested for flow cytometry. (C and D) To assess whether CEACAM5 is important for MERS-CoV attachment, human CEACAM5-expressing BHK21 cells were challenged with MERS-CoV at an MOI of 30 for 2 h at 4°C before being harvested for flow cytometry. Human DPP4-expressing BHK21 cells were included as controls for both entry and attachment assays. (B and D) The data are represented as the percentages of MERS-CoV NP-positive BHK21 cells after infection with or without DPP4/CEACAM5 overepression. The means and standard deviations were derived from the results of three independent experiments. Statistical analyses were carried out using Student's t test. Statistical significance is indicated by the asterisks ( P

    Journal: Journal of Virology

    Article Title: Carcinoembryonic Antigen-Related Cell Adhesion Molecule 5 Is an Important Surface Attachment Factor That Facilitates Entry of Middle East Respiratory Syndrome Coronavirus

    doi: 10.1128/JVI.01133-16

    Figure Lengend Snippet: CEACAM5 is an attachment factor for MERS-CoV. (A and B) To assess whether CEACAM5 is important for MERS-CoV entry, human CEACAM5 was overexpressed in BHK21 cells. Human CEACAM5-expressing BHK21 cells were then challenged with MERS-CoV at an MOI of 5 for 2 h at 37°C. The inoculum was replaced with culture medium, and the cells were incubated for another 4 h before being harvested for flow cytometry. (C and D) To assess whether CEACAM5 is important for MERS-CoV attachment, human CEACAM5-expressing BHK21 cells were challenged with MERS-CoV at an MOI of 30 for 2 h at 4°C before being harvested for flow cytometry. Human DPP4-expressing BHK21 cells were included as controls for both entry and attachment assays. (B and D) The data are represented as the percentages of MERS-CoV NP-positive BHK21 cells after infection with or without DPP4/CEACAM5 overepression. The means and standard deviations were derived from the results of three independent experiments. Statistical analyses were carried out using Student's t test. Statistical significance is indicated by the asterisks ( P

    Article Snippet: MERS-CoV spike was detected either with the in-house mouse anti-MERS-CoV spike immune serum or with a rabbit anti-MERS-CoV spike antibody (Sino Biological, China).

    Techniques: Expressing, Incubation, Flow Cytometry, Cytometry, Infection, Derivative Assay

    CEACAM5-specific antibody blocks MERS-CoV entry and replication. (A) The antibody blocking assay was performed in Calu3 cells using MERS-S-pseudotyped virus. Antibodies were diluted to 2 μg/ml and incubated with Calu3 cells for 1 h at 37°C. The pseudotyped viruses were then added at a ratio of 100 LP per cell for 1 h. Luciferase activity was determined at 48 h postinfection and was normalized to that of the mock-treated cells. (B) The antibody blocking assay was performed in Calu3 cells using MERS-CoV. Calu3 cells were preincubated with antibodies at the indicated concentrations for 1 h at 37°C. The cells were then inoculated with MERS-CoV at an MOI of 1 for 1 h at 37°C in the presence of the antibodies. After 1 h, the cells were washed and harvested. MERS-CoV entry was assessed by qPCR, and the result was normalized to that of the mock-treated cells. (C) Calu3 cells were treated with CEACAM5 antibody for a total of 2 h during preincubation and virus inoculation (−1 to 1) or after virus inoculation (1 to 3). MERS-CoV entry was assessed by qPCR. (D) The antibody blocking assay was performed in Caco2 cells. (E and F) The impact of CEACAM5 inhibition on MERS-CoV replication was investigated in Huh7 cells. Huh7 cells were preincubated with antibodies at 2 μg/ml for 1 h at 37°C. The cells were then inoculated with MERS-CoV at an MOI of 0.0005 for 1 h at 37°C in the presence of the antibodies. At the end of the inoculation period, the inoculum was replaced with culture medium containing the indicated antibodies. Cell lysates (E) and supernatants (F) were harvested at 1, 24, and 48 h postinfection. The virus genome copy number was determined with qPCR. ND, virus was not detected. The data are represented as means and standard deviations of the results of three independent experiments. The results for the anti-DPP4-, anti-CEACAM5-, and control IgG-treated samples were compared with those for the mock-treated samples. Statistical analyses were carried out using Student's t test. Statistical significance is indicated by the asterisks ( P

    Journal: Journal of Virology

    Article Title: Carcinoembryonic Antigen-Related Cell Adhesion Molecule 5 Is an Important Surface Attachment Factor That Facilitates Entry of Middle East Respiratory Syndrome Coronavirus

    doi: 10.1128/JVI.01133-16

    Figure Lengend Snippet: CEACAM5-specific antibody blocks MERS-CoV entry and replication. (A) The antibody blocking assay was performed in Calu3 cells using MERS-S-pseudotyped virus. Antibodies were diluted to 2 μg/ml and incubated with Calu3 cells for 1 h at 37°C. The pseudotyped viruses were then added at a ratio of 100 LP per cell for 1 h. Luciferase activity was determined at 48 h postinfection and was normalized to that of the mock-treated cells. (B) The antibody blocking assay was performed in Calu3 cells using MERS-CoV. Calu3 cells were preincubated with antibodies at the indicated concentrations for 1 h at 37°C. The cells were then inoculated with MERS-CoV at an MOI of 1 for 1 h at 37°C in the presence of the antibodies. After 1 h, the cells were washed and harvested. MERS-CoV entry was assessed by qPCR, and the result was normalized to that of the mock-treated cells. (C) Calu3 cells were treated with CEACAM5 antibody for a total of 2 h during preincubation and virus inoculation (−1 to 1) or after virus inoculation (1 to 3). MERS-CoV entry was assessed by qPCR. (D) The antibody blocking assay was performed in Caco2 cells. (E and F) The impact of CEACAM5 inhibition on MERS-CoV replication was investigated in Huh7 cells. Huh7 cells were preincubated with antibodies at 2 μg/ml for 1 h at 37°C. The cells were then inoculated with MERS-CoV at an MOI of 0.0005 for 1 h at 37°C in the presence of the antibodies. At the end of the inoculation period, the inoculum was replaced with culture medium containing the indicated antibodies. Cell lysates (E) and supernatants (F) were harvested at 1, 24, and 48 h postinfection. The virus genome copy number was determined with qPCR. ND, virus was not detected. The data are represented as means and standard deviations of the results of three independent experiments. The results for the anti-DPP4-, anti-CEACAM5-, and control IgG-treated samples were compared with those for the mock-treated samples. Statistical analyses were carried out using Student's t test. Statistical significance is indicated by the asterisks ( P

    Article Snippet: MERS-CoV spike was detected either with the in-house mouse anti-MERS-CoV spike immune serum or with a rabbit anti-MERS-CoV spike antibody (Sino Biological, China).

    Techniques: Antibody Blocking Assay, Incubation, Luciferase, Activity Assay, Real-time Polymerase Chain Reaction, Inhibition

    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: Then, human anti-MERS-S2P antibodies (S2A3, S2A6, and S2D5) and a rabbit polyclonal anti-MERS-CoV spike protein antibody, used as a positive control (Sino biological), in TBS (0.1% Triton X-100, 1% BSA in PBS) solution were incubated with the Vero cells infected with MERS-CoV.

    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: Then, human anti-MERS-S2P antibodies (S2A3, S2A6, and S2D5) and a rabbit polyclonal anti-MERS-CoV spike protein antibody, used as a positive control (Sino biological), in TBS (0.1% Triton X-100, 1% BSA in PBS) solution were incubated with the Vero cells infected with MERS-CoV.

    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: Then, human anti-MERS-S2P antibodies (S2A3, S2A6, and S2D5) and a rabbit polyclonal anti-MERS-CoV spike protein antibody, used as a positive control (Sino biological), in TBS (0.1% Triton X-100, 1% BSA in PBS) solution were incubated with the Vero cells infected with MERS-CoV.

    Techniques: Enzyme-linked Immunosorbent Assay, Sandwich ELISA