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

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.

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

Supplementary files

Article information

Article type
Paper
Submitted
08 Apr 2025
Accepted
25 May 2025
First published
26 May 2025
This article is Open Access
Creative Commons BY license

Analyst, 2025, Advance Article

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

M. Wallace, J. M. Sharpe, K. Baj, M. Ngwube, J. Thirlway, P. L. Kerigan Higgs, G. R. Stephenson, J. A. Iggo, T. E. Storr and C. J. Richards, Analyst, 2025, Advance Article , DOI: 10.1039/D5AN00406C

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