Issue 22, 2024

Impact of drug-linker on method selection for analytical characterization and purification of antibody–drug conjugates

Abstract

In addition to traditional characterisation methods of hydrophobic interaction (HIC) and reverse phase (RP) chromatography, an anion exchange chromatography (AIEX) was developed to analyse and purify antibody drug conjugates (ADCs). Since different drug antibody ratio (DAR) species may impact biological activity, therapeutic index, PK parameters or even potential immunogenicity, homogenous ADC DAR demands have been significantly increasing. To accelerate linker designs, drug screening and ADC DAR purification for in vitro and in vivo studies, we built the analytical toolbox including HIC, RP, AIEX, icIEF, SEC, and MS for downstream ADC DAR purification using HIC and AIEX. The established analytical methods can quickly assess the quality of ADC DAR profiles and provide important information to select the proper ADC DAR purification method. Since drug-linker structures can significantly affect ADC physicochemical properties, and highly impact on selections of analytical methods, we applied both HIC and AIEX characterisation and purification platforms to achieve ADC DAR homogenous. Our experiments also implied that unlike HIC, AIEX could be used to separate DAR4 positional isomers.

Graphical abstract: Impact of drug-linker on method selection for analytical characterization and purification of antibody–drug conjugates

Supplementary files

Article information

Article type
Paper
Submitted
20 Apr 2024
Accepted
16 May 2024
First published
16 May 2024

Anal. Methods, 2024,16, 3492-3503

Impact of drug-linker on method selection for analytical characterization and purification of antibody–drug conjugates

L. C. Santora, A. D. Hobson, L. Wang and K. X. Wu, Anal. Methods, 2024, 16, 3492 DOI: 10.1039/D4AY00725E

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