Evaluation of High-Affinity Monoclonal Antibodies and Antibody-Drug Conjugates by Homogenous Time-Resolved FRET

Authors

Harmon Greenway
Jin Wang

Language

English

Publication Date

9-12-2024

Journal

ACS Medical Chemistry Letters Journal

DOI

10.1021/acsmedchemlett.4c00317

PMID

39291004

PMCID

PMC11403740

PubMedCentral® Posted Date

8-30-2024

PubMedCentral® Full Text Version

Post-print

Abstract

The rapid growth of therapeutic monoclonal antibodies demands greater accessibility to scalable methods of evaluating antigen binding. Homogenous TR-FRET is ideal for preliminary screening but has not been reported to assay these interactions due to their high-affinity and complex solution-phase kinetics. Here we report the development of a competition assay to rank-order the relative affinities of these drugs for a common antigen. The assay is compatible with automation, requires no modification of the analytes, and measures affinities as low as single-digit picomolar. We further demonstrate applications to inform the development of antibody-drug conjugates. The assay may aid discovery and manufacturing of therapeutic antibodies as a low-cost, high-throughput alternative to existing technologies.

Keywords

Binding Affinity, Monoclonal Antibodies, Antibody-Drug Conjugates, Homogenous TR-FRET

Comments

This article has been corrected. See ACS Med Chem Lett. 2024 Nov 29;15(12):2231.

Published Open-Access

yes

Recommended Citation

Harmon Greenway and Jin Wang, "Evaluation of High-Affinity Monoclonal Antibodies and Antibody-Drug Conjugates by Homogenous Time-Resolved FRET" (2024). Faculty and Staff Publications. 3859.
https://digitalcommons.library.tmc.edu/baylor_docs/3859