Journal: PLoS Pathogens

Article Title: Human coronaviruses disassemble processing bodies

doi: 10.1371/journal.ppat.1010724

Figure Lengend Snippet: Plasmids.

Article Snippet: Codon-optimized pLJM1-MERS-CoV-N-FLAG was cloned from SinoBiological (cat #VG40068-CF) using BamHI and EcoRI.

Techniques: Selection, Plasmid Preparation, Over Expression

Journal: Cell Host & Microbe

Article Title: A Potent Germline-like Human Monoclonal Antibody Targets a pH-Sensitive Epitope on H7N9 Influenza Hemagglutinin

doi: 10.1016/j.chom.2017.08.011

Figure Lengend Snippet: The Schematic Biopanning Process and the Characterization of H7 HA-Specific mAb from the Antibody Library (A) The biopanning strategy of H7 HA1-specific mAbs from the naive antibody phage display library. (B) Polyclonal phage ELISA showing the binding of the first to fifth rounds (Rd 1 to Rd 5) of phages to HA and HA1. Bound phages were detected with anti-M13-HRP conjugate. (C) Binding of Fab m826 to four different subtypes of influenza virus HA. The error bars reflect the SD. (D) Immunogenetic analysis of the heavy- and light-chain variable regions of m826 using the IMGT tool. (E) Germline-rooted circular phylogenetic tree of m826-like antibody sequences found in IgM libraries derived from healthy human adults, neonates, and H7N9-infected patients. The sequences of m826 and the clone identical to m826 are shown in red. Sequence ID started with CB represents sequences derived from the neonates, started with HH represents sequences derived from the healthy adults, and started with H7N9 represents that from the H7N9-infected patients. The phylogenetic tree was constructed by the Neighbor-Joining method.

Article Snippet: pCMV3-H7N9-HA-GFPSpark , Sinobiological Inc. , Cat# VG40103-ACG.

Techniques: Enzyme-linked Immunosorbent Assay, Binding Assay, Derivative Assay, Infection, Sequencing, Construct

Journal: Cell Host & Microbe

Article Title: A Potent Germline-like Human Monoclonal Antibody Targets a pH-Sensitive Epitope on H7N9 Influenza Hemagglutinin

doi: 10.1016/j.chom.2017.08.011

Figure Lengend Snippet: Binding Profiles of m826 mAb to HA and HA1 Measured by BLI in OctetRED96 The m826 mAb was immobilized on activated AR2G biosensors. The analytes consisted of serial dilution (between 100 and 1.2 nM) of trimeric H7N9 HA at pH 5.0 (A) or pH 7.4 (B), H7N9 HA1 at pH 7.4 (C), or 100 nM H1N1 and H3N2 HA at pH 7.4 (D). Binding kinetics was evaluated using a 1:1 Langmuir binding model by ForteBio Data Analysis 7.0 software.

Article Snippet: pCMV3-H7N9-HA-GFPSpark , Sinobiological Inc. , Cat# VG40103-ACG.

Techniques: Binding Assay, Serial Dilution, Software

Journal: Cell Host & Microbe

Article Title: A Potent Germline-like Human Monoclonal Antibody Targets a pH-Sensitive Epitope on H7N9 Influenza Hemagglutinin

doi: 10.1016/j.chom.2017.08.011

Figure Lengend Snippet: Analysis of Detailed Interactions between the m826 Fab and H7N9 HA (A and B) The crystal structures of Fab m826 in complex with H7N9 HA1; view from the side (A) and the top (B). (C) Close-up of the three CDRs of heavy chain from the m826 bound to HA. Side and main chains involved in hydrogen bonding are shown as sticks, with hydrogen bonds depicted as black dotted lines. Residues of the HA protein are labeled. CDR regions are shown as tubes. (D) The three CDRs of light chain also make direct contacts with HA to form several hydrogen bonds (black dashed lines) with the HA residues. (E) Amino acid sequences of m826 are shown with Kabat numbering. The CDRs are colored and amino acids involved in HA binding are labeled with ∗ .

Article Snippet: pCMV3-H7N9-HA-GFPSpark , Sinobiological Inc. , Cat# VG40103-ACG.

Techniques: Labeling, Binding Assay

Journal: Cell Host & Microbe

Article Title: A Potent Germline-like Human Monoclonal Antibody Targets a pH-Sensitive Epitope on H7N9 Influenza Hemagglutinin

doi: 10.1016/j.chom.2017.08.011

Figure Lengend Snippet: m826 mAb Recognizes a Unique Epitope on H7 HA (A) Docking of the m826-HA1 complex onto the crystal structure of monomeric H7 HA. The epitope of m826 is distinct from the RBS. (B) Superposition of the antibody epitope which was partially buried in the trimeric HA structure. (C) Sequence alignment of HA1 from the H7N9, H7N7, and H7N4, and highlighting of structurally defined influenza antibody epitopes. The conventional antigenic sites (Sa, Sb, Ca1, Ca2, and Cb) are shaded in blue, the epitope of m826 is shaded in red, and RBS is shown in the black box.

Article Snippet: pCMV3-H7N9-HA-GFPSpark , Sinobiological Inc. , Cat# VG40103-ACG.

Techniques: Sequencing

Journal: Cell Host & Microbe

Article Title: A Potent Germline-like Human Monoclonal Antibody Targets a pH-Sensitive Epitope on H7N9 Influenza Hemagglutinin

doi: 10.1016/j.chom.2017.08.011

Figure Lengend Snippet: m826 mAb Does Not Neutralize H7 Virus but Mediates Potent ADCC (A) The immunostaining-based neutralization assay for m826 mAb against H7N4 virus. Immunostaining was conducted using influenza NP-specific mAbs to detect virus-infected cells. H7N4-specfic Ab was used as positive control. (B) Confocal images of pCMV3-H7N9-HA-GFP transfected 293T cells stained with 100 nM m826 IgG or an unrelated anti-hepatitis B virus human IgG G12 as the control. Green signal, GFP; red signal, detecting with Alexa Fluor 647-conjugated goat anti-human IgG. Scale bar, 25 μm. (C) ADCC-related signaling in Jurkat T cells engineered to express human FcγRIIIa triggered by m826 and CR9114 IgG in the presence of H7N9 HA-expressing 293T cells. An unrelated anti-MERS-CoV IgG m336 was used as negative control. (D) ADCC activity of m826 and CR9114 IgG against H7N9 HA-expressing 293T cells mediated by freshly prepared human PBMCs. The percentage (%) of specific lysis was calculated as described in the . All error bars reflect SD.

Article Snippet: pCMV3-H7N9-HA-GFPSpark , Sinobiological Inc. , Cat# VG40103-ACG.

Techniques: Immunostaining, Neutralization, Infection, Positive Control, Transfection, Staining, Expressing, Negative Control, Activity Assay, Lysis

Journal: Cell Host & Microbe

Article Title: A Potent Germline-like Human Monoclonal Antibody Targets a pH-Sensitive Epitope on H7N9 Influenza Hemagglutinin

doi: 10.1016/j.chom.2017.08.011

Figure Lengend Snippet: Prophylactic and Therapeutic Efficacy of mAb m826 in Protecting Mice against Lethal Dose H7N9 Challenge (A–D) For prophylactic efficacy study, mice were treated (intraperitoneally [i.p.]) with m826 antibody 12 hr before viral challenge (intranasally [i.n.]) with 10 × LD 50 of H7N9 virus and were monitored daily for 14 days for the accumulated mortality (A) and weight loss (B), expressed as percent (%) weight loss and survival, respectively (n = 5 per group). PBS was used for the control group. For therapeutic efficacy study, mice were treated (i.p.) with m826 antibody 12 hr after viral challenge (i.n.) with 10 × LD 50 of H7N9 virus and were monitored daily for 14 days for the accumulated mortality (C) and weight loss (D). The weight loss of (B and D) represents mean change in body weight per group and SD was also shown. (E) Evaluation of histopathological changes in the pulmonary tissues of uninfected mice and mice from therapeutic, prophylactic, and control group. The black, yellow, and blue arrows represented the H7N9 infection-induced inflammation of blood vessel, alveoli, or capillary, respectively. (F) Evaluation of histopathological changes in the bronchial tissues. The black, yellow, blue, and green arrows represented bronchial epithelial cell, bronchial alveoli, capillary, and interstitial tissue, respectively.

Article Snippet: pCMV3-H7N9-HA-GFPSpark , Sinobiological Inc. , Cat# VG40103-ACG.

Techniques: Infection

Journal: Cell Host & Microbe

Article Title: A Potent Germline-like Human Monoclonal Antibody Targets a pH-Sensitive Epitope on H7N9 Influenza Hemagglutinin

doi: 10.1016/j.chom.2017.08.011

Figure Lengend Snippet:

Article Snippet: pCMV3-H7N9-HA-GFPSpark , Sinobiological Inc. , Cat# VG40103-ACG.

Techniques: Recombinant, Electron Microscopy, Affinity Chromatography, Integrity Assay, Expressing, Plasmid Preparation, Software

Journal: Microbiology Spectrum

Article Title: Rapid and Quantitative Detection of Human Antibodies against the 2019 Novel Coronavirus SARS CoV2 and Its Variants as a Result of Vaccination and Infection

doi: 10.1128/Spectrum.00890-21

Figure Lengend Snippet: (A) Human IgG levels against SARS CoV2 antigens, measured by GC-FP, for vaccinated individuals versus unvaccinated individuals. The Mann-Whitney test was used to determine statistical significance. (B) Human IgG levels against SARS CoV2 antigens throughout the vaccination sequence (Pfizer-BioNTech) for 10 subjects, collected prevaccination, at the time of the 2nd dose of vaccine, and 2 weeks after the 2nd dose of vaccine. One-way analysis of variance (ANOVA) followed by Dunnett’s multiple-comparison testing was performed (*, P = 0.03; **, P = 0.002; ***, P = 0.0002; ****, P < 0.0001).

Article Snippet: SARS CoV2 S1—B.1.351/Beta (K417N, E484K, N501Y, D614G) , Sino Biological.

Techniques: MANN-WHITNEY, Sequencing

Journal: Microbiology Spectrum

Article Title: Rapid and Quantitative Detection of Human Antibodies against the 2019 Novel Coronavirus SARS CoV2 and Its Variants as a Result of Vaccination and Infection

doi: 10.1128/Spectrum.00890-21

Figure Lengend Snippet: IgG levels against SARS CoV2 antigens for uninfected, previously infected, and vaccinated individuals. Uninfected samples were collected prior to vaccination, from individuals who reported no prior COVID-19 symptoms, and tested negative via PCR and/or antibody testing ( n = 42). Other samples were from PCR-confirmed COVID-19-positive subjects who were hospitalized ( n = 3), PCR-confirmed COVID-19-positive subjects who were not hospitalized CoV2 ( n = 5), and previously COVID-19-positive subjects who received subsequent vaccination ( n = 9). Samples were also collected from subjects who were at least 2 weeks past full vaccination with Pfizer-BioNTech ( n = 17) or Moderna ( n = 8) or 2 weeks past receiving the Johnson & Johnson vaccine ( n = 9). One-way ANOVA followed by Dunnett’s multiple-comparison test was performed (*, P = 0.03; **, P = 0.002; ***, P = 0.0002; ****, P < 0.0001).

Article Snippet: SARS CoV2 S1—B.1.351/Beta (K417N, E484K, N501Y, D614G) , Sino Biological.

Techniques: Infection

Journal: Microbiology Spectrum

Article Title: Rapid and Quantitative Detection of Human Antibodies against the 2019 Novel Coronavirus SARS CoV2 and Its Variants as a Result of Vaccination and Infection

doi: 10.1128/Spectrum.00890-21

Figure Lengend Snippet: Fold-difference in antibody levels against antigens from SARS CoV2 variant strains B.1.1.7 and B.1.351 versus antigens from the original 2019 SARS CoV2 strain. Samples included those from individuals who were hospitalized ( n = 3), nonhospitalized CoV2-positive ( n = 5), previously CoV2-positive with subsequent vaccination ( n = 9), and at least 2 weeks past vaccination with Pfizer-BioNTech ( n = 17), Moderna ( n = 8), or Johnson & Johnson vaccine ( n = 9). One-way ANOVA followed by Dunnett’s multiple-comparison test was performed (*, P = 0.03; **, P = 0.002; ***, P = 0.0002; ****, P < 0.0001).

Article Snippet: SARS CoV2 S1—B.1.351/Beta (K417N, E484K, N501Y, D614G) , Sino Biological.

Techniques: Variant Assay

Journal: Microbiology Spectrum

Article Title: Rapid and Quantitative Detection of Human Antibodies against the 2019 Novel Coronavirus SARS CoV2 and Its Variants as a Result of Vaccination and Infection

doi: 10.1128/Spectrum.00890-21

Figure Lengend Snippet: (A and B) IgG levels from dried blood spots measured by GC-FP diagnostic ratio compared to competitive ELISA by eluate from the same dried blood spot samples. Both GC-FP and ACE2 competitive binding were performed for RBD antigen from the original 2019 SARS CoV2 and the variant strains B.1.1.7 and B.1.351. Testing was performed with dried blood spots collected from vaccinated subjects (3 Pfizer-BioNTech, 3 Moderna, 2 Johnson & Johnson) and subjects who were both previously infected and then vaccinated with Pfizer-BioNTech or Moderna vaccines ( n = 3). One-way ANOVA followed by Dunnett’s multiple-comparison test was performed (*, P = 0.03; **, P = 0.002; ***, P = 0.0002; ****, P < 0.0001). (C and D) The percentage of ACE2 binding inhibition from the competitive ELISA is shown for a dried blood spot sample (C) from a Pfizer-BioNTech-vaccinated subject and blood serum from a hospitalized, COVID-positive subject (D). Vertical dotted lines represent the dilution factor used in the corresponding GC-FP test for each sample. (E and F) The fold difference in binding inhibition and fold difference in GC-FP diagnostic ratio were plotted for variant antigens (RBD B.1.1.7 and RBD B.1.351) versus RBD 2019 CoV2.

Article Snippet: SARS CoV2 S1—B.1.351/Beta (K417N, E484K, N501Y, D614G) , Sino Biological.

Techniques: Diagnostic Assay, Competitive ELISA, Binding Assay, Variant Assay, Infection, Inhibition

Journal: Microbiology Spectrum

Article Title: Rapid and Quantitative Detection of Human Antibodies against the 2019 Novel Coronavirus SARS CoV2 and Its Variants as a Result of Vaccination and Infection

doi: 10.1128/Spectrum.00890-21

Figure Lengend Snippet: Proteins and peptides used for generating GC-FP detection microchips

Article Snippet: SARS CoV2 S1—B.1.351/Beta (K417N, E484K, N501Y, D614G) , Sino Biological.

Techniques: Positive Control, Negative Control