human fcγri  (R&D Systems)

Journal: Frontiers in Immunology

Article Title: A novel monoclonal antibody against human thymic stromal lymphopoietin for the treatment of TSLP-mediated diseases

doi: 10.3389/fimmu.2024.1442588

Figure Lengend Snippet: TSLP binding and epitope competition studies for TAVO101 and tezepelumab. (A) . Increasing concentrations of TAVO101 and tezepelumab were assessed for their binding to immobilized human TSLP. OD at 450 nm were plotted against the concentrations of test antibodies (Data expressed as mean ± SEM, n=2). (B) . Increasing concentrations of TAVO101 and tezepelumab were assessed for their inhibition on the binding of human TSLP to its receptor complex expressed on the surface of HEK293T cells transfected with IL7Rα and TSLPR by flow cytometry. Mean fluorescence intensities (MFI) representing TSLP binding were plotted against the concentrations of test antibodies (Data expressed as mean ± SEM, n=3). (C) . Epitope competition binding assays for TAVO101 and tezepelumab by gator Bio-layer Interferometry. After loading the biosensor with biotinylated TSLP during the first binding phase, either TAVO101 (left panel) or tezepelumab (right panel) was added until the binding to the TSLP on the biosensor was saturated during the second phase. In the third binding phase, either tezepelumab or TAVO101 was then added to monitor kinetic binding to the TSLP on the biosensor. Shifts in interference reflecting the binding of test antibodies to TSLP were plotted against time (in second) during the three phases of binding. In each assay, triplicate binding profiles for the competition antibody addition and a single binding profile for the same antibody addition during the third binding phase were shown.

Article Snippet: 100 nM of biotinylated recombinant human FcγRI (CD64) or FcγRIIIA (CD16a) (AcroBiosystems, Newark, DE) were coated on 96-well plate pre-coated with streptavidin (Sigma, St. Louis, MO).

Techniques: Binding Assay, Inhibition, Transfection, Flow Cytometry, Fluorescence

Journal: Frontiers in Immunology

Article Title: A novel monoclonal antibody against human thymic stromal lymphopoietin for the treatment of TSLP-mediated diseases

doi: 10.3389/fimmu.2024.1442588

Figure Lengend Snippet: Fc engineering of TAVO101 for extended half-life and reduced Fcγ receptor binding. (A) . Binding to mouse FcRn by Fc engineered TAVO101. Increasing concentrations of TAVO101_IgG1-AALS and TAVO101_IgG1-FEA were assessed for their binding to immobilized mouse FcRn at pH 6.0 in ELISA assays. OD at 450 nm were plotted against the concentrations of test antibodies (Data expressed as mean ± SEM, n=2). (B) . Pharmacokinetic profile of an Fc engineered TAVO101. TAVO101_IgG1-AALS antibody was administered as a single 4 mg/kg intravenous dose into a cynomolgus monkey. The antibody concentrations in plasma at time points up to day 35 post-dose were quantitated and plotted against the testing days. The calculated pharmacokinetic parameters were shown. (C) . Binding to human Fcγ receptors and C1q complement protein by Fc engineered TAVO101. Increasing concentrations of TAVO101_IgG1-AALS antibody and a human IgG1 control antibody were assessed for their binding to FcγRI (CD64), FcγRIIIA (CD16a), and C1q. OD at 450 nm were plotted against the concentrations of test antibodies (Data expressed as mean ± SEM, n=3).

Article Snippet: 100 nM of biotinylated recombinant human FcγRI (CD64) or FcγRIIIA (CD16a) (AcroBiosystems, Newark, DE) were coated on 96-well plate pre-coated with streptavidin (Sigma, St. Louis, MO).

Techniques: Binding Assay, Enzyme-linked Immunosorbent Assay, Clinical Proteomics, Control

Journal: PLOS Pathogens

Article Title: Polyfunctionality and breadth of HIV-1 antibodies are associated with delayed disease progression

doi: 10.1371/journal.ppat.1012739

Figure Lengend Snippet: Polyfunctionality was assessed by calculation of a polyfunctionality score at 3 years post-seroconversion (SC). We only assessed activating FcγR (FcγRI, FcγRIIa, FcγRIIIa, FcγRIIIb) and C1q interaction, and for each Fc feature we determined for each participant if there was a response that was higher than the median response of all responders in the cohort. The resulting scores (amount of features above the responder median, between 0 and 5, per person) were plotted for the four groups based on time between SC and acquired immunodeficiency syndrome (AIDS) (less than 3 years (n = 28), between 3 and 7 years (n = 147), between 7 and 11 years (n = 66) and more than 11 years (n = 71)) with one plot for 92BR020 gp120-specific responses and one plot for 92BR020 SOSIP-specific responses. In addition, a polyfunctionality index was calculated for each group as a quantitative measure of polyfunctionality. This is a value between 0 and 100 calculated by a weighted addition of the percentage of individuals in each category based on the number of features, derived from Larsen et al . , see ). This index is shown above each bar. Polyfunctionality of antibodies against these two strains at 6 months post-SC are shown in .

Article Snippet: Human FcγRI and FcγRIIIb proteins were acquired from Invitrogen and human purified C1q protein was acquired from Complement Technologies.

Techniques: Derivative Assay

Journal: Experimental Hematology & Oncology

Article Title: Preclinical investigations and a first-in-human phase 1a trial of JS007, a novel anti-CTLA-4 antibody, in patients with advanced solid tumors

doi: 10.1186/s40164-024-00567-7

Figure Lengend Snippet: JS007 binding affinity to CTLA-4 protein and Fc receptor by Biacore

Article Snippet: The affinity of JS007 for Fc receptors was determined using Biacore (GE Healthcare Life Sciences), in which anti-His tag antibody was coupled to a CM5 chip surface, followed by capturing the His-tagged recombinant human FcγRIIIa (CD16a) V176 (Junmeng Biomedical, 20200421), FcγRIIIa (CD16a) F176 (Junmeng Biomedical, 20200421), FcγRIIa (CD32a) R167 (Sino Biological, 10734-H08C), FcγRI (CD64) (Sino Biological, 10256-H08S), and FcRn (Sino Biological, CT009-H08H).

Techniques: Binding Assay, Recombinant