Journal: ACS Omega

Article Title: Click Chemistry-Generated Auristatin F–Linker–Benzylguanine for a SNAP-Tag-Based Recombinant Antibody–Drug Conjugate Demonstrating Selective Cytotoxicity toward EGFR-Overexpressing Tumor Cells

doi: 10.1021/acsomega.2c06844

Figure Lengend Snippet: Live cell imaging of 1711(scFv)-SNAP-Alexa Fluor 647 binding to EGFR-positive cells. (A–D) Confocal microscopy images of 1711(scFv)-SNAP-Alexa Fluor 647 binding and uptake (red) in target cell lines after 30 min of incubation at room temperature. Hoechst was used as a stain for the nuclei (blue). (A) Surface binding of 1711(scFv)-SNAP-Alexa Fluor 647 to A431 cells at 37 °C. (B) Specific surface binding and internalization of 1711(scFv)-SNAP-Alexa Fluor 647 in MDA-MB-468 cells at room temperature. (C) No binding of 1711(scFv)-SNAP-Alexa Fluor 647 to the EGFR-negative A2058 cells. (D) Vehicle control with MDA-MB-468 cells incubated in 1× PBS at room temperature.

Article Snippet: MDA-MB-468 (ATCC: HTB-132), A431 (ATCC: CRL-259), and A2058 (ATCC: CRL-11147) cells were cultured in Dulbecco’s modified Eagle’s medium (DMEM) (containing 2 mM L-glutamine, 3.7 g/L C 3 H 3 NaO 3 and 16 mg/L phenol red) supplemented as above.

Techniques: Live Cell Imaging, Binding Assay, Confocal Microscopy, Incubation, Staining, Control