Programmable wide-range pH gradients for NMR titrations: Application to antibody–drug conjugate linker group modifications

Matthew Wallace, James M. Sharpe, Krzysztof Baj, Michael Ngwube, Jenny Thirlway, Patrick L. Kerigan Higgs, G. Richard Stephenson, Jonathan A. Iggo, Thomas E. Storr, Christopher J. Richards

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Abstract

We generate pH gradients spanning more than six units in standard NMR tubes and determine all the pKa values of polyprotic compounds in single 20 minutes chemical shift imaging (CSI) NMR experiments. The modest demands of our method in terms of sample quantity and preparation time allow it be performed as part of the routine characterisation and optimisation of organic molecules during synthesis campaigns. As proof of concept, we measure the pKa values of a family of vinylpyridines employed as antibody drug conjugate linkers. Our analysis reveals a strong correlation between the experimental aqueous pKa and the rate of conjugate addition of thiol nucleophiles to the vinyl group, representing a powerful predictive method.
Original languageEnglish
Pages (from-to)2872-2879
Number of pages8
JournalAnalyst
Volume150
Issue number13
Early online date26 May 2025
DOIs
Publication statusPublished - 7 Jul 2025

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