One-pot thiol–amine bioconjugation to maleimides: simultaneous stabilisation and dual functionalisation
Archie Wall, Alfie Wills, Nafsika Forte, Calise Bahou, Lisa Bonin, Karl Nicholls, Michelle Ma, Vijay Chudasama, James R. Baker
Abstract
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.