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UPLC-Based Assay to Assess the Hydrophobicity of Antibody-Drug Conjugate (ADC) Payloads

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Original languageEnglish
Article numberj.jchromb.2020.122075
Number of pages7
JournalJournal of Chromatography B: Analytical Technologies in the Biomedical & Life Sciences
Accepted/In press19 Mar 2020

King's Authors


Antibody-Drug Conjugates (ADCs) are comprised of antibodies attached to cytotoxic small molecules or biological agents (i.e., payloads) through chemical linkers which may be cleavable or non-cleavable. The development of new ADCs is challenging, particularly the process of attaching the linker-payload construct to the antibody (i.e., the conjugation process). One of the major problems associated with conjugation is high hydrophobicity of the payload which can lead to low yields of the ADC through aggregation and/or lower than desired Drug-Antibody Ratios (DARs). We report here a UPLC-based assay that can be used to study the physicochemical properties of ADC payloads at an early stage of development, and to provide information on whether the hydrophilic/hydrophobic balance is suitable for conjugation or whether further physicochemical optimization is required. The assay is relatively simple to establish and should be of use to those working in the ADC area.

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