Abstract
This chapter describes an LC-ESI-MS method for the DAR and drug load distribution analysis that is suitable for lysine-linked ADCs. The ADC sample is desalted using a reversed-phase LC column with an acetonitrile gradient prior to online MS analysis. The MS spectrum is processed (deconvoluted) and converted to a series of zero charge state masses that corresponds to the increasing number of drugs in the ADC. Integration of the mass peak area allows the calculation of the DAR and drug load distribution of ADCs.
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Acknowledgements
We thank Pat Rancatore, Richard Seipert, Boyan Zhang, and Andrea Ji for comments and help in preparing this review.
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Basa, L. (2013). Drug-to-Antibody Ratio (DAR) and Drug Load Distribution by LC-ESI-MS. In: Ducry, L. (eds) Antibody-Drug Conjugates. Methods in Molecular Biology, vol 1045. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-541-5_18
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DOI: https://doi.org/10.1007/978-1-62703-541-5_18
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