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
