An ex situ gaseous reagent for multicomponent amine bioconjugation

Abstract

Bioconjugation is a large field with many diverse goals, needs, and challenges, that requires a broad toolbox of fundamentally different synthetic approaches. As nucleophilic groups are prevalent in biomolecules, the ability to crosslink two nucleophilic sites offers an attractive approach to construct useful bioconjugates. New technologies for crosslinking with gaseous reagents and with minimal perturbation of natural structure could provide new ways to think about bioconjugation in complex environments. We report a minimalist gaseous sulfonyl chloride-derived reagent for multicomponent bioconjugation with amine, phenol, or aniline reagents to afford urea or carbamate products. In utilizing a gas-phase reagent for a reaction mediated by metal ions, a variety of biologically relevant molecules such as saccharide, PEG, fluorophore, and affinity tag can be efficiently crosslinked to the N-terminus or lysine side chain amines on natural polypeptides or proteins. The application of this method to the production of functional, modified proteins was demonstrated by fluorescence imaging of a cancer cell line and by the facile preparation of a peptide–protein conjugate.

Ex Situ Gaseous Reagent for Multicomponent Amine Bioconjugation

“…We discovered , this reactivity while investigating carbonylative coupling, in which CO was generated in a two-chambered reactor for safe ex situ production of CO . When CO was produced from formic acid and methanesulfonyl chloride with triethylamine, bioconjugation reactions of amine reagents persisted, even in negative control experiments without formic acid .…”