One-Pot Thiol-Amine Bioconjugation to Maleimides; Simultaneous Stabilisation and Dual Functionalisation

12 June 2020, Version 1
This content is a preprint and has not undergone peer review at the time of posting.


Maleimide chemistry is widely used in the site-selective modification of proteins. However, hydrolysis of the resultant thiosuccinimides is required to provide robust stability to the bioconjugates. Herein, we present an alternative approach that affords simultaneous stabilisation and dual functionalisation in a one pot fashion. By consecutive conjugation of a thiol and an amine to dibromomaleimides, we show that aminothiomaleimides can be generated extremely efficiently. Furthermore, the amine serves to deactivate the electrophilicity of the maleimide, precluding further reactivity and hence generating stable conjugates. We have applied this conjugation strategy to peptides and proteins to generate stabilised trifunctional conjugates. We propose that this stabilisation-dual modification strategy could have widespread use in the generation of diverse conjugates.


protein modification reactions
maleimide derivatives
albumin conjugates
Dual conjugation

Supplementary materials

Amine-Thiol Dual Conjugation ESI


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