Journal: Journal of Virological Methods

Article Title: Multiplexed, microscale, microarray-based serological assay for antibodies against all human-relevant coronaviruses

doi: 10.1016/j.jviromet.2021.114111

Figure Lengend Snippet: Precision, Accuracy of all 9 CoV SeroAssay Capture Antigens. a

Article Snippet: Fluorescence microarray images illustrating binding of monoclonal antibodies to the CoV SeroAssay. (a) CR3022 SARS-CoV-1 antibody from Creative Biolabs binding to the nCoV(ii) and SARS antigens, (b) 40021-MM07 HKU1 antibody from Sino Biological binding to HKU1 antigen, (c) 40069-MM23 MERS antibody from Sino Biological binding to the MERS antigen, and (d) GTX632604 SARS-CoV-2 antibody from Genetex binding to the nCoV(i) and nCoV(iii) antigens. (a) Schematic illustration of microarray layout, and representative fluorescence images of the VaxArray CoV SeroAssay microarray in (b) through (l).

Techniques: Concentration Assay

Journal: Journal of Virological Methods

Article Title: Multiplexed, microscale, microarray-based serological assay for antibodies against all human-relevant coronaviruses

doi: 10.1016/j.jviromet.2021.114111

Figure Lengend Snippet: Identifying Information for Nine Human Coronavirus Spike Antigens Represented on the CoV SeroAssay.

Article Snippet: Fluorescence microarray images illustrating binding of monoclonal antibodies to the CoV SeroAssay. (a) CR3022 SARS-CoV-1 antibody from Creative Biolabs binding to the nCoV(ii) and SARS antigens, (b) 40021-MM07 HKU1 antibody from Sino Biological binding to HKU1 antigen, (c) 40069-MM23 MERS antibody from Sino Biological binding to the MERS antigen, and (d) GTX632604 SARS-CoV-2 antibody from Genetex binding to the nCoV(i) and nCoV(iii) antigens. (a) Schematic illustration of microarray layout, and representative fluorescence images of the VaxArray CoV SeroAssay microarray in (b) through (l).

Techniques: Expressing, Binding Assay

Journal: Journal of Virological Methods

Article Title: Multiplexed, microscale, microarray-based serological assay for antibodies against all human-relevant coronaviruses

doi: 10.1016/j.jviromet.2021.114111

Figure Lengend Snippet: Sensitivity, Linear Dynamic Range of all 9 CoV SeroAssay Capture Antigens.

Article Snippet: Fluorescence microarray images illustrating binding of monoclonal antibodies to the CoV SeroAssay. (a) CR3022 SARS-CoV-1 antibody from Creative Biolabs binding to the nCoV(ii) and SARS antigens, (b) 40021-MM07 HKU1 antibody from Sino Biological binding to HKU1 antigen, (c) 40069-MM23 MERS antibody from Sino Biological binding to the MERS antigen, and (d) GTX632604 SARS-CoV-2 antibody from Genetex binding to the nCoV(i) and nCoV(iii) antigens. (a) Schematic illustration of microarray layout, and representative fluorescence images of the VaxArray CoV SeroAssay microarray in (b) through (l).

Techniques:

Journal: Journal of Virological Methods

Article Title: Multiplexed, microscale, microarray-based serological assay for antibodies against all human-relevant coronaviruses

doi: 10.1016/j.jviromet.2021.114111

Figure Lengend Snippet: Fluorescence microarray images illustrating binding of monoclonal antibodies to the CoV SeroAssay. (a) CR3022 SARS-CoV-1 antibody from Creative Biolabs binding to the nCoV(ii) and SARS antigens, (b) 40021-MM07 HKU1 antibody from Sino Biological binding to HKU1 antigen, (c) 40069-MM23 MERS antibody from Sino Biological binding to the MERS antigen, and (d) GTX632604 SARS-CoV-2 antibody from Genetex binding to the nCoV(i) and nCoV(iii) antigens.

Article Snippet: Fluorescence microarray images illustrating binding of monoclonal antibodies to the CoV SeroAssay. (a) CR3022 SARS-CoV-1 antibody from Creative Biolabs binding to the nCoV(ii) and SARS antigens, (b) 40021-MM07 HKU1 antibody from Sino Biological binding to HKU1 antigen, (c) 40069-MM23 MERS antibody from Sino Biological binding to the MERS antigen, and (d) GTX632604 SARS-CoV-2 antibody from Genetex binding to the nCoV(i) and nCoV(iii) antigens. (a) Schematic illustration of microarray layout, and representative fluorescence images of the VaxArray CoV SeroAssay microarray in (b) through (l).

Techniques: Fluorescence, Microarray, Binding Assay

Journal: The Lancet. Infectious Diseases

Article Title: T-cell responses to MERS coronavirus infection in people with occupational exposure to dromedary camels in Nigeria: an observational cohort study

doi: 10.1016/S1473-3099(20)30599-5

Figure Lengend Snippet: MERS-CoV-specific CD4 + and CD8 + T-cell responses in camel workers and controls (A) Frequencies of MERS-CoV-specific CD4+ T cells. (B) Frequencies of MERS-CoV-specific CD8+ T cells. (C) Summary of aggregate CD4+ T-cell responses to all structural peptide pools in different study groups. (D) Summary of aggregate CD8+ T-cell responses to all structural peptide pools in different study groups. (E) CD4+ T-cell responses to MERS-CoV accessory protein-specific peptide pools. (F) Phenotypes of virus-specific CD4+ T cells. (G,H) Phenotypes of virus-specific CD8+ T cells. Abattoir workers with exposure to dromedaries are represented by red symbols, those without exposure to dromedaries by green symbols, non-abattoir workers by light blue symbols, MERS-positive controls by dark blue symbols (open shapes represent asymptomatic patients), and negative controls from Guangzhou by purple symbols. Symbol shape identifies the same individual. IFN=interferon. MERS-CoV= Middle East respiratory syndrome coronavirus. TNF=tumour necrosis factor. **=p<0·01. ***=p<0·001.

Article Snippet: Anti-MERS-CoV antibody titres were determined using plaque reduction neutralisation tests., A set of 20-mer peptides overlapping by ten amino acids encompassing the four MERS-CoV (HCoV-EMC/2012) structural proteins (peptides S1, S2, N, and ME encompassing the N-terminal and C-terminal portions of the spike [S] glycoprotein, the nucleocapsid [N] protein, and the transmembrane [M] and envelope [E] proteins) and five accessory proteins (ORF3, ORF4a, ORF4b, ORF5 and ORF8b) were synthesised by Sino Biological (Shanghai, China), and used for stimulation of PBMCs.

Techniques:

Journal: Nature Communications

Article Title: Characterization of spike glycoprotein of SARS-CoV-2 on virus entry and its immune cross-reactivity with SARS-CoV

doi: 10.1038/s41467-020-15562-9

Figure Lengend Snippet: a Diagram of full-length SARS-CoV-2 S protein with a 3xFLAG tag. S1, receptor-binding subunit; S2, membrane fusion subunit; TM, transmembrane domain; NTD, N-terminal domain; pFP, potential fusion peptide; HR-N, heptad repeat-N; HR-C, heptad repeat-C; b – f Detection of CoVs S protein in cells lysate by western blot. Mock, 293T cells transfected with empty vector. b Mouse monoclonal anti-FLAG M2 antibody; c Polyclonal goat anti-MHV-A59 S protein antibody AO4. d Polyclonal rabbit anti-SARS S1 antibodies T62. e Mouse monoclonal anti-SARS S1 antibody. f Mouse monoclonal anti-MERS-CoV S2 antibody. g – j Detection of CoVs S protein in pseudovirions by western blot.Gag-p24 served as a loading control. g Anti-FLAG M2. h Polyclonal goat anti-MHV-A59 S protein antibody AO4. i Polyclonal rabbit anti-SARS S1 antibodies T62. j Polyclonal anti-Gag-p24 antibodies. uncleaved S protein, about 180 kDa; cleaved S protein, about 90 kDa. Experiments were done twice and one is shown. Source data are provided as a Source Data file.

Article Snippet: Rabbit polyclonal against SARS S1 antibodies (#40150-T62), mouse monoclonal against MERS-CoV S2 antibody (#40070-MM11), mouse monoclonal against SARS S1 antibody (#40150-MM02), rabbit polyclonal against HIV-1 Gag-p24 antibody (11695-RP01) were purchased from Sino Biological Inc. (Beijing, China).

Techniques: Binding Assay, Western Blot, Transfection, Plasmid Preparation

Journal: Nature Communications

Article Title: Characterization of spike glycoprotein of SARS-CoV-2 on virus entry and its immune cross-reactivity with SARS-CoV

doi: 10.1038/s41467-020-15562-9

Figure Lengend Snippet: a , b Entry of SARS-CoV-2 S pseudovirions on indicated cell lines. Cells from human and animal origin were inoculated with SARS-CoV-2 S (red), SARS-CoV S (blue), or VSV-G (gray) pseudovirions. At 48 h post inoculation, transduction efficiency was measured according to luciferase activities. RS, Rhinolophus sinicus bat embryonic fibroblast; BHK/hAPN, BHK cells stably expressing hAPN, the hCoV-229E receptor; 293/hACE2, 293 cells stably expressing hACE2, the SARS-CoV receptor; HeLa/hDPP4, HeLa cells stably expressing hDPP4, the MERS-CoV receptor. Experiments were done in triplicates and repeated at least three times. One representative is shown with error bars indicating SEM. c Binding of SARS-CoV S and SARS-CoV-2 S proteins to soluble hACE2. HEK293T cells transiently expressing SARS-CoV and SARS-CoV-2 S proteins were incubated with the soluble hACE2 on ice, followed by polyclonal goat anti-hACE2 antibody. Cells were analyzed by flow cytometry. The experiments were repeated at least three times. d Inhibition of SARS-CoV-2 S pseudovirion entry by soluble hACE2. SARS-CoV S, SARS-CoV-2 S, or VSV-G pseudovirions were pre-incubated with soluble hACE2, then mixture were added to 293/hACE2 cells. Cells were lysed 40 h later and pseudoviral transduction was measured. Experiments were done twice and one representative is shown. Error bars indicate SEM of technical triplicates. Source data are provided as a Source Data file.

Article Snippet: Rabbit polyclonal against SARS S1 antibodies (#40150-T62), mouse monoclonal against MERS-CoV S2 antibody (#40070-MM11), mouse monoclonal against SARS S1 antibody (#40150-MM02), rabbit polyclonal against HIV-1 Gag-p24 antibody (11695-RP01) were purchased from Sino Biological Inc. (Beijing, China).

Techniques: Transduction, Luciferase, Stable Transfection, Expressing, Binding Assay, Incubation, Flow Cytometry, Inhibition

Journal: Nature Communications

Article Title: Characterization of spike glycoprotein of SARS-CoV-2 on virus entry and its immune cross-reactivity with SARS-CoV

doi: 10.1038/s41467-020-15562-9

Figure Lengend Snippet: a Inhibition of entry of SARS-CoV-2 S pseudovirion on 293/hACE2 by lysosomotropic agents (20 mM NH 4 Cl and 100 nM bafilomycin A). b Inhibition of entry of SARS-CoV, MERS-CoV, and MHV S pseudovirions by a PIKfyve inhibitor apilimod. HeLa/mCEACAM, 293/hACE2, HeLa/hDPP4 cells were pretreated with different concentrations of apilimod and transduced with MHV S, SARS-CoV S, MERS-CoV S pseudovirions, respectively. The luciferase activity was measured 40 h post transduction. VSV-G pseudovirions were used as a control. Experiments were done in triplicates and repeated at least three times. One representative is shown with error bars indicating SEM. c Inhibition of MHV A59 infection by apilimod. The 17Cl.1 cells were pretreated with 3, 10, 30, 100, 300 nM apilimod for 30 min and infected by MHV A59 at MOI = 0.01. Viral infection and cell viability were determined by using qPCR and MTT assay, respectively. Experiments were done in triplicates and repeated at least three times. One representative is shown with error bars indicating SEM. d , e Inhibition of entry of SARS-CoV-2 S protein pseudovirions by apilimod, YM201636, and tetrandrine. HEK 293/hACE2 cells were pretreated with either apilimod ( d ), YM201636 ( e ), or tetrandrine ( f ), then inoculated with SARS-CoV-2 S pseudovirons in the presence of drug. The luciferase activity were measured 40 h post transduction. YM201636, PIKfyve inhibitor; tetrandrine, TPC2 inhibitor. The experiments were done in triplicates and repeated at least three times. One representative is shown with error bars indicating SEM of technical triplicates. Source data are provided as a Source Data file.

Article Snippet: Rabbit polyclonal against SARS S1 antibodies (#40150-T62), mouse monoclonal against MERS-CoV S2 antibody (#40070-MM11), mouse monoclonal against SARS S1 antibody (#40150-MM02), rabbit polyclonal against HIV-1 Gag-p24 antibody (11695-RP01) were purchased from Sino Biological Inc. (Beijing, China).

Techniques: Inhibition, Transduction, Luciferase, Activity Assay, Infection, MTT Assay

Journal: Mikrochimica Acta

Article Title: Detection of SARS-CoV-2 receptor binding domain using fluorescence probe and DNA flowers enabled by rolling circle amplification

doi: 10.1007/s00604-023-05747-6

Figure Lengend Snippet: Evaluation of specificity of the RCA-enabled fluorometric ( a ) and colorimetric assay ( b ). The signal intensity generated from S protein RBD is compared with that from N protein, MERS protein RBD, SARS-Cov-2 B.1.617.2 spike protein RBD, and a mixture of the three proteins. The total protein concentration in each sample was 50 ng/mL

Article Snippet: Recombinant SARS-CoV-2 receptor-binding domain (RBD, source: HEK293-derived SARS-CoV-2 Spike RBD protein (Arg319-Phe541), with a 6x-His tag at C-terminal), recombinant MERS-CoV Spike RBD (Chinese Hamster Ovary-derived MERS-CoV Spike RBD protein (Glu367-Tyr606), with a 6x-His tag at C-terminal), recombinant SARS-CoV-2 B.1.617.2 RBD (source: HEK293-derived SARS-CoV-2 Spike RBD protein (Arg319-Phe541 (Leu452Arg, Thr478Lys), with a 6x-His tag at C-terminal), and recombinant SARS-CoV-2 nucleocapsid (source: Spodoptera frugiperda , Sf 21 (baculovirus)-derived SARS-CoV-2 nucleocapsid (Met1-Ala419), with a 6x-His tag at C-terminal) were purchased from Bio-Techne (Minneapolis, USA).

Techniques: Colorimetric Assay, Generated, Protein Concentration

Journal: bioRxiv

Article Title: Analysis of Serologic Cross-Reactivity Between Common Human Coronaviruses and SARS-CoV-2 Using Coronavirus Antigen Microarray

doi: 10.1101/2020.03.24.006544

Figure Lengend Snippet: Coronavirus antigens on microarray.

Article Snippet: Coronavirus , MERS , MERS , S1-RBD , AFS88936.1 , Insect Cells , Sino Biological , N-(AA383-502)-Fc-C , 40071-V05B.

Techniques: Microarray, Expressing, Construct

Journal: Science Bulletin

Article Title: Binding of the SARS-CoV-2 spike protein to glycans

doi: 10.1016/j.scib.2021.01.010

Figure Lengend Snippet: A possible mechanism for SARS-CoV-2 entry and infection. At the early stage of the infection process, SARS-CoV-2 may first interact with the HSPGs on the surface of susceptible cells using the S protein protruding from the virus particle. This initial attachment may promote the subsequent binding of the virus to the high-affinity entry receptor ACE2. The transmembrane protease serine 2 (TMPRSS2) on host cell surface and other host cell proteases may assist in viral entry by cleaving the S protein at the S1/S2 and/or at the S2’ sites.

Article Snippet: The recombinant SARS-CoV-2-S2 (686–1213), MERS-CoV-RBD (367–606), MERS-CoV-S1 (1–725), MERS-CoV-S2 (726–1296), and MERS-CoV-S (1-1297) with C-terminal His-tags were purchased from Sino Biological (Beijing, China).

Techniques: Infection, Binding Assay

Journal: Science Bulletin

Article Title: Binding of the SARS-CoV-2 spike protein to glycans

doi: 10.1016/j.scib.2021.01.010

Figure Lengend Snippet: Schematic representation of CoV S proteins. The diagrams show the domain organization of the S proteins of SARS-CoV-2, SARS-CoV, and MERS-CoV. The sequences of the S1/S2 and S2’ cleavage sites are given on top of each diagram. The arrows indicate the cleave points.

Article Snippet: The recombinant SARS-CoV-2-S2 (686–1213), MERS-CoV-RBD (367–606), MERS-CoV-S1 (1–725), MERS-CoV-S2 (726–1296), and MERS-CoV-S (1-1297) with C-terminal His-tags were purchased from Sino Biological (Beijing, China).

Techniques:

Journal: Science Bulletin

Article Title: Binding of the SARS-CoV-2 spike protein to glycans

doi: 10.1016/j.scib.2021.01.010

Figure Lengend Snippet: The binding of full-length CoV S proteins and their domains to HS microarrays. (a) SARS-CoV-2-RBD binding at the concentration of 2.5 μg/mL, (b) SARS-CoV-2-S1 at the concentration of 2 μg/mL, (c) SARS-CoV-2-S2 at the concentration of 2 μg/mL, (d) SARS-CoV-2-S at the concentration of 2 μg/mL, (e) SARS-CoV-2-trimer at the concentration of 2 μg/mL, (f) SARS-CoV-RBD at the concentration of 2 μg/mL, (g) SARS-CoV-S1 at the concentration of 2 μg/mL, (h) SARS-CoV-2-S at the concentration of 2 μg/mL, (i) MERS-CoV-RBD at the concentration of 4 μg/mL, (j) MERS-CoV-S1 at the concentration of 4 μg/mL, (k) MERS-CoV-S2 at the concentration of 2 μg/mL, (l) MERS-CoV-S at the concentration of 2 μg/mL. The fluorescence intensity was measured at an excitation wavelength of 532 nm. All error bars are standard deviation of more than three replicates.

Article Snippet: The recombinant SARS-CoV-2-S2 (686–1213), MERS-CoV-RBD (367–606), MERS-CoV-S1 (1–725), MERS-CoV-S2 (726–1296), and MERS-CoV-S (1-1297) with C-terminal His-tags were purchased from Sino Biological (Beijing, China).

Techniques: Binding Assay, Concentration Assay, Fluorescence, Standard Deviation

Journal: Science Bulletin

Article Title: Binding of the SARS-CoV-2 spike protein to glycans

doi: 10.1016/j.scib.2021.01.010

Figure Lengend Snippet: SPR sensorgrams of the interactions between a commercially available porcine heparin and immobilized S proteins and their domains. Panels (a−l) show the binding of heparin to SARS-CoV-2-RBD, S1, S2, S, trimer, SARS-CoV-RBD, S1, S, and MERS-CoV-RBD, S1, S2, S, respectively. The name of each protein and its K D value are presented on top of the corresponding sensorgram. RU, resonance unit.

Article Snippet: The recombinant SARS-CoV-2-S2 (686–1213), MERS-CoV-RBD (367–606), MERS-CoV-S1 (1–725), MERS-CoV-S2 (726–1296), and MERS-CoV-S (1-1297) with C-terminal His-tags were purchased from Sino Biological (Beijing, China).

Techniques: Binding Assay

Journal: Science Bulletin

Article Title: Binding of the SARS-CoV-2 spike protein to glycans

doi: 10.1016/j.scib.2021.01.010

Figure Lengend Snippet: SPR sensorgrams of the interactions between fondaparinux and immobilized S proteins and their domains. Panels (a–l) are the binding of fondaparinux to SARS-CoV-2-RBD, S1, S2, S, trimer, SARS-CoV-RBD, S1, S, and MERS-CoV-RBD, S1, S2, S, respectively. The name and K D value of each protein are presented on top of the corresponding sensorgram. RU, resonance unit.

Article Snippet: The recombinant SARS-CoV-2-S2 (686–1213), MERS-CoV-RBD (367–606), MERS-CoV-S1 (1–725), MERS-CoV-S2 (726–1296), and MERS-CoV-S (1-1297) with C-terminal His-tags were purchased from Sino Biological (Beijing, China).

Techniques: Binding Assay