The stability of antibody-drug conjugates (ADCs) in circulation is dictated by a number of factors, one of which is the susceptibility of the linker to premature cleavage by circulating esterases and proteases. Indeed, both esterases (such as CES1C) and proteases (such as neutrophil elastase) are speculated to play a significant role in both the PK-exposure and toxicity of several ADCs that have been evaluated in the clinic. With this in mind, we have designed a library of ADC linkers that can easily be screened by a FRET-based assay against various enzymes and biological media. The library consists of ~75 tripeptides connected to a fluorophore-quencher pair on opposing ends of the peptide. Cleavage of the peptide results in an increase in fluorescence that is easily observed in a 96-well plate-based assay format. We will describe the design of the library, the optimization of the assay, and the screening that has resulted in the identification of peptides that may offer increased utility over existing cathepsin-B cleavable ADC linkers.

Evaluation of a FRET-based peptide-library aimed at the identification of improved proteolytic substrates for cleavable linkers

Macromolecular Chemistry: The Second Century

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ACS Spring 2020 National Meeting & Expo

The official archive and repository for all ACS Meetings & Expositions abstracts and select posters or presentations.