Journal: Frontiers in Oncology

Article Title: A trispecific antibody targeting EGFR/cMET/VEGF-A demonstrates multiple mechanisms of action to inhibit wild-type and mutant NSCLC animal models

doi: 10.3389/fonc.2025.1533059

Figure Lengend Snippet: TAVO412 bound to NSCLC cell lines and blocked binding of EGF and HGF. The binding of TAVO412 (red open circle), Amivantamab analogue (blue open circle) and null mAb (black open circle) to (A) NCI-H292; (B) HCC827; and (C) NCI-H1975 NSCLC cell lines as analyzed by flow cytometry. See Supplementary Table S2 for corresponding EC 50 , 95% CI for EC 50 , and efficacy (span in y axis). Blocking of (D) EGF and (E) HGF from binding to HCC827 cells was assessed by flow cytometry with TAVO412 (red open circle), Amivantamab analogue (blue open circle) and null mAb (black open circle). See Supplementary Table S3 for corresponding IC 50 , 95% CI for IC 50 , and efficacy (span in Y-axis). The data from three independent experiments were expressed as the mean ± SEM of duplicate treatments. The amivantamab analogue served as a positive control molecule while the null mAb served as a negative control. The abbreviations were: gMFI, geometric mean fluorescent intensity; AF647, Alexa Fluor 647 dye; AF488, Alexa Fluor 488 dye; Ab, antibody; nM, nanomolar; EGF, epidermal growth factor; HGF, hepatocyte growth factor; SEM, standard error of the mean.

Article Snippet: The cells were then incubated with AF647 goat anti-human IgG1 Fc (Jackson ImmunoResearch, 109-605-190) in the dark for 30 min at 4°C, washed three times with FACS buffer, and resuspended in FACS buffer for flow cytometry (Beckman CytoFLEX) experiments.

Techniques: Binding Assay, Flow Cytometry, Blocking Assay, Positive Control, Negative Control