Background: Antibody–drug conjugates (ADCs) are a new class of cancer therapeutics that deliver potent cytotoxins specifically to tumors to minimize systemic toxicity. However, undesirable release of covalently linked drugs in circulation can affect safety and efficacy. The objective of this manuscript was to propose and assess the assays that allow for the characterization of the drug deconjugation in plasma/serum. Results: ADCs of three main drug conjugation platforms, linked via lysine, site-specific engineered cysteine or reduced interchain disulfide cysteine residues, were analyzed using affinity capture for sample enrichment coupled with LC–MS or hydrophobic interaction chromatography–UV for detection. These novel approaches enabled measurement of the relative abundance of individual ADC species with different drug-to-antibody ratios, while maintaining their structural integrity. Conclusion: The characterization data generated by affinity capture LC–MS or hydrophobic interaction chromatography–UV provided critical mechanistic insights into understanding the stability and bioactivity of ADCs in vivo, and also helped the development of appropriate quantitative ELISAs.

Papers of special note have been highlighted as: ▪ of interest ▪▪ of considerable interest

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Vol. 5, No. 9

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Published online 3 May 2013

Published in print May 2013

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Acknowledgements

The authors would like to thank the following individuals for helpful discussions: O Saad, C Ng, N Koppada, B Shen, J Tibbitts, F Jacobson, J Gorell and P Polakis. The authors also want to thank G Deperalta, S Mahrus and A Song for their critical review of the manuscript.

Financial & competing interests disclosure

All of the authors are employees of Genentech. The authors have no other relevant affiliations or financial involvement with any organization or entity with a financial interest in or financial conflict with the subject matter or materials discussed in the manuscript apart from those disclosed.

No writing assistance was utilized in the production of this manuscript.

Ethical conduct of research

The authors state that they have obtained appropriate institutional review board approval or have followed the principles outlined in the Declaration of Helsinki for all human or animal experimental investigations. In addition, for investigations involving human subjects, informed consent has been obtained from the participants involved.

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