Bispecific anti-mPDGFR beta x cotinine scFv-C-kappa-scFv fusion protein and cotinine-duocarmycin can form antibody-drug conjugate-like complexes that exert cytotoxicity against mPDGFR beta expressing cells

Abstract

Antibody selection for antibody-drug conjugates (ADCs) has traditionally depended on its internalization into the target cell, although ADC efficacy also relies on recycling of the receptor-ADC complex, endo-lysosomal trafficking, and subsequent linker/antibody proteolysis. In this study, we observed that a bispecific anti-murine platelet-derived growth factor receptor beta (mPDGFR beta) x cotinine single-chain variable fragment (scFv)-kappa constant region (C-kappa)-scFv fusion protein and cotinine-duocarmycin can form an ADC-like complex to induce cytotoxicity against rnPDGFR beta expressing cells. Multiple anti-mPDGFR beta antibody candidates can be produced in this bispecific scFv-C-kappa-scFv fusion protein format and tested for their ability to deliver cotinine-conjugated cytotoxic drugs, thus providing an improved approach for antibody selection in ADC development.