Journal: mAbs

Article Title: Preclinical evaluation of AFM24, a novel CD16A-specific innate immune cell engager targeting EGFR-positive tumors

doi: 10.1080/19420862.2021.1950264

Figure Lengend Snippet: Inhibition of EGF-stimulated EGFR phosphorylation by AFM24. A) A-431 or B) A-549 cells were incubated with increasing concentrations of either AFM24 or comparators (RSV/CD16A scFv-IgAb, EGFR/RSV scFv-IgAb, or cetuximab) and subsequently stimulated with 100 ng/mL recombinant human EGF for 10 min at 37°C. Cell lysates were used in an ELISA for the detection of phosphorylated EGFR. Phosphorylation intensity was normalized to EGF-stimulated samples without antibody incubation. Error bars represent SD. One representative experiment is shown out of seven (for A-431) or two (A-549) performed

Article Snippet: EGF (100 ng/mL; Thermo Fisher Scientific, cat.: 10605-HNAE-250) was added for 10 min at 37°C before cells were washed with ice-cold PBS, lysed and used for relative quantification of phosphorylated EGFR using a Phospho-EGFR ELISA Kit (enzyme-linked immunosorbent assays, RayBiotech, cat.: PEL-EGFR-Y) according to the manufacturer’s instructions.

Techniques: Inhibition, Incubation, Recombinant, Enzyme-linked Immunosorbent Assay