Author ORCID Identifier

Document Type


Date of Award

Spring 4-22-2020

Degree Name

Biochemistry (BS)



First Advisor

Dr. L. Nathan Tumey

Second Advisor

Dr. Susan C. Flynn

Third Advisor

Dr. Ming An


Science and Mathematics

Subject Heading(s)

Antibody-drug conjugates; Bioconjugates; Amino acids


Success of Antibody-drug conjugates (ADCs) relies on the maintained integrity of a chemical linker fusing a therapeutic payload to a monoclonal antibody. Recent reports have revealed the lead linker ValCitPABC has poor stability in rodent models compared to mammalian counterparts, resulting in many clinical investigations predicting poorer ADC efficacy due to premature payload release in these model systems. Optimization of this chemical linker to be resistant both in mouse and human models would streamline ADC progression into clinical trials. Herein we describe the synthesis and development of a FRET-based assay for evaluating linker stability in vitro. Evaluation of lysosomal release by catabolic proteases paired with serum stability testing identified a series of asparagine containing linkers which surpassed both the release kinetics and stability profiles of ValCitPABC. A synthetic scheme for MMAE attachment to these asparagine linkers was established. Ultimately, this work lays a foundation for future evaluation of these linker in vivo for the identification of next generation peptide linkers to surpass the limitations of ValCitPABC.

Included in

Biochemistry Commons