Cetuximab DM1 Antibody-Drug Conjugate Cytotoxicity with Lung Cancer Cell Lines

Abstract

The goal of the study was to create an antibody-drug conjugate which would be potentially more effective than conventional chemotherapeutic treatments. Through DNA transduction a plasmid was introduced to Escherichia coli (E. coli). The plasmid containing E. coli was selected with hygromycin. The plasmid was then extracted and purified from E. coli then grown in TB broth. Expi293TM cells were then transfected with the purified DNA to create the Cetuximab antibody. Then through a fluorescein label, the binding affinity of Cetuximab to various cancer cell lines were determined through flow cytometry. The two highest bindings were A549 and HCC827 and the lowest binding was H2170. The three cell lines were selected for further tests. Through the conjugation of Cetuximab with SMCC-DM1 the antibody drug conjugate of Cetuximab-DM1 (Cet-DM1) was created at the Drug-to-Antibody ratio of 7.5:1, 15:1, and 30:1. From cytotoxicity assays it was determined the Cet-DM1 Drug-to-Antibody ratio of 15:1 was the best to continue experimentation. The IC50 of the Cet-DM1 15:1 ADC with A549 cells was 4.113e-2 mg/mL and with H2170 cell was 8.284e-3 mg/mL. This indicated that the Cet-DM1 15:1 ADC was more cytotoxic than naked Cetuximab and more selective than just DM1 for A549 and H2170 cell lines.