Journal: Cell Reports Medicine

Article Title: Therapeutic activity of an inhaled potent SARS-CoV-2 neutralizing human monoclonal antibody in hamsters

doi: 10.1016/j.xcrm.2021.100218

Figure Lengend Snippet: Isolation of SARS-CoV-2 RBD-specific human monoclonal antibodies (hmAbs) (A) Representative gating strategy for RBD + B cell isolation. Initial plots are gated on live CD3 − CD4 − CD19 + annexin V-CD27 + B cells. (B) hmAbs were tested at 10 and 1 μg/mL in duplicate by ELISA for binding to indicated protein; area under the curve (AUC) is indicated. (C) hmAbs were tested at 10 μg/mL in duplicate for binding to indicated protein by ELISA in the presence or absence of 8 M urea treatment, mean and standard error presented. (D) Binding to SARS-CoV-2 RBD was determined by surface plasmon resonance (SPR), mean and standard deviation presented.

Article Snippet: SARS-CoV-2 S1+S2 , Sino Biological , Cat#40589-V08B1-B.

Techniques: Isolation, Cell Isolation, Enzyme-linked Immunosorbent Assay, Binding Assay, SPR Assay, Standard Deviation

Journal: Cell Reports Medicine

Article Title: Therapeutic activity of an inhaled potent SARS-CoV-2 neutralizing human monoclonal antibody in hamsters

doi: 10.1016/j.xcrm.2021.100218

Figure Lengend Snippet: Neutralization of SARS-CoV-2 infection and RBD binding to ACE2 (A) Indicated hmAbs were incubated with live SARS-CoV-2 (100 PFU/well) 1 h before (pre-treatment) or 1 h after (post-treatment) addition to Vero E6 cells. hmAbs were tested in quadruplicate cultures and NT 50 and upper 95% confidence interval (CI) indicated. (B) Representative titration curve of 1212C2 hmAb presented, mean and standard error presented. (C) Binding of indicated hmAb to SARS-CoV-2 or mock-infected Vero E6 cells measured by immunofluorescence; scale bar, 100 μm. (D) Indicated hmAb was incubated as single replicate with recombinant biotinylated RBD protein before incubation with HEK293-ACE2 cells measured by flow cytometry. Plot gated on 7-aminoactinomycin (7AAD)-ACE2 + cells.

Article Snippet: SARS-CoV-2 S1+S2 , Sino Biological , Cat#40589-V08B1-B.

Techniques: Neutralization, Infection, Binding Assay, Incubation, Titration, Immunofluorescence, Recombinant, Flow Cytometry

Journal: Cell Reports Medicine

Article Title: Therapeutic activity of an inhaled potent SARS-CoV-2 neutralizing human monoclonal antibody in hamsters

doi: 10.1016/j.xcrm.2021.100218

Figure Lengend Snippet: SPR epitope mapping (A) Representative sensorgram from the SPR competition assays used to subset hmAbs into distinct RBD binding epitopes. For each assay, a series of hmAbs were sequentially injected over immobilized SARS-CoV-2 RBD. In this example, 1212C2 was injected first, followed by a second injection of 1212C2, 2 injections of 1206D1, and the last injection was CR3022. (B) Summary of all epitope mapping data, in which each block (first experiment from A in the red box) with a bold hmAb at the top represents a different experiment (10 experiments total). The bold hmAb is the “first” hmAb injected. The percentage of binding (100 = 100% binding and 0 = 0% binding) of subsequent hmAbs was recorded. mAbs were considered to have a different epitope (denoted by a distinct color) if they exhibited binding levels >30% in the presence of other mAbs. Thus, in the first experiment, CR3022 is defined as a new epitope (cyan), distinct from 1212C2. (C) Schematic diagram of NmAb RBD epitopes defined in the mapping experiment. Five major epitopes (A–E) were identified, where the E epitope overlaps with control mAb CR3022 (cyan, epitope F). Four of the 5 epitopes (A–D) are located within the ACE2 binding site (purple), and all of the NmAbs are blocked by the 1212C2 epitope A (yellow). NmAbs with epitopes similar to B (orange) and C (green) are defined as B’ (light orange) and C’ (light green), respectively. The 1212C2 epitope A (yellow) blocks the binding of all NmAbs, with the exception of 1215B11, which occupies epitope E. Epitopes B and C are also blocked by epitope A NmAbs, but exhibit limited competition with each other.

Article Snippet: SARS-CoV-2 S1+S2 , Sino Biological , Cat#40589-V08B1-B.

Techniques: Binding Assay, Injection, Blocking Assay

Journal: Cell Reports Medicine

Article Title: Therapeutic activity of an inhaled potent SARS-CoV-2 neutralizing human monoclonal antibody in hamsters

doi: 10.1016/j.xcrm.2021.100218

Figure Lengend Snippet: Prophylactic and therapeutic activity of 1212C2 hmAb in SARS-CoV-2-infected hamsters (A and B) Golden Syrian hamsters were (A) prophylactically treated i.p. with 10 mg/kg indicated hmAb or PBS 6 h before intranasal (i.n.) challenge with 2 × 10 5 PFU SARS-CoV-2 or (B) therapeutically treated i.p. with 25 mg/kg indicated hmAb or PBS 6 h following i.n. challenge with SARS-CoV-2. Virus present in nasal turbinates and lungs was determined by plaque assay. Dotted line indicates limit of detection. (C and D) Distribution of pathologic lesion, including consolidation, congestion, and pneumonic lesions, was measured using ImageJ and represented as the percentage of the total lung surface area in prophylactically treated (C) and therapeutically treated (D) animals. Each symbol represents an individual animal. p values determined by 2-tailed t test.

Article Snippet: SARS-CoV-2 S1+S2 , Sino Biological , Cat#40589-V08B1-B.

Techniques: Activity Assay, Infection, Plaque Assay

Journal: Cell Reports Medicine

Article Title: Therapeutic activity of an inhaled potent SARS-CoV-2 neutralizing human monoclonal antibody in hamsters

doi: 10.1016/j.xcrm.2021.100218

Figure Lengend Snippet: Therapeutic activity of inhaled 1212C2 hmAb in SARS-CoV-2-infected hamsters (A and B) Golden Syrian hamsters were infected i.n. with 2 × 10 5 PFU SARS-CoV-2 and 12 h later treated with indicated hmAb by i.p. or IH administration. Virus present in nasal turbinates and lungs was determined by plaque assay at 2 dpi (A) and 4 dpi (B). Dotted line indicates limit of detection. (C) Distribution of pathologic lesion were measured using ImageJ and represented as the percentage of the total lung surface area. ∗p < 0.0055 and ∗∗p < 0.00055 compared to PBS i.p.-treated group determined by 2-tailed t test.

Article Snippet: SARS-CoV-2 S1+S2 , Sino Biological , Cat#40589-V08B1-B.

Techniques: Activity Assay, Infection, Plaque Assay

Journal: Cell Reports Medicine

Article Title: Therapeutic activity of an inhaled potent SARS-CoV-2 neutralizing human monoclonal antibody in hamsters

doi: 10.1016/j.xcrm.2021.100218

Figure Lengend Snippet:

Article Snippet: SARS-CoV-2 S1+S2 , Sino Biological , Cat#40589-V08B1-B.

Techniques: Synthesized, Expressing, Recombinant, Plasmid Preparation, Binding Assay, Transfection, Gel Extraction, Lysis, Luciferase, Software