UPLC-based assay to assess the hydrophobicity of Antibody-Drug Conjugate (ADC) payloads.

Ilona Pysz, Paul J M Jackson, David J Barlow, Khondaker Miraz Rahman, David E Thurston

Journal: Journal of chromatography. B, Analytical technologies in the biomedical and life sciences 2020;1146():122075

PMID: 32361467

Full Text on Elsevier Science View on PubMed

Abstract

Antibody-Drug Conjugates (ADCs) consist 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 further physicochemical optimization is required. The assay is relatively simple to establish and should be of use to those working in the ADC area.

Address: Institute of Pharmaceutical Science, Faculty of Life Sciences & Medicine, King's College London, Franklin-Wilkins Building, 150 Stamford Street, London SE1 9NH, United Kingdom; Femtogenix Ltd, Lawes Open Innovation Hub, Rothamsted Research, West Common, Harpenden, Hertfordshire AL5 2JQ, United Kingdom.; Femtogenix Ltd, Lawes Open Innovation Hub, Rothamsted Research, West Common, Harpenden, Hertfordshire AL5 2JQ, United Kingdom.; Institute of Pharmaceutical Science, Faculty of Life Sciences & Medicine, King's College London, Franklin-Wilkins Building, 150 Stamford Street, London SE1 9NH, United Kingdom.; Institute of Pharmaceutical Science, Faculty of Life Sciences & Medicine, King's College London, Franklin-Wilkins Building, 150 Stamford Street, London SE1 9NH, United Kingdom; Femtogenix Ltd, Lawes Open Innovation Hub, Rothamsted Research, West Common, Harpenden, Hertfordshire AL5 2JQ, United Kingdom. Electronic address: [email protected].

DOI: 10.1016/j.jchromb.2020.122075

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MeSH Terms: Biological Assay, Calicheamicins, Chromatography, High Pressure Liquid, Doxorubicin, Flurbiprofen, Hydrophobic and Hydrophilic Interactions, Ibuprofen, Immunoconjugates, Irinotecan, Ketoprofen, Maytansine, Molecular Conformation, Norfloxacin, Pentachlorophenol, Protein Multimerization, Structure-Activity Relationship, Tandem Mass Spectrometry, Tolnaftate