Fiche publication
Date publication
janvier 2023
Journal
Methods in molecular biology (Clifton, N.J.)
Auteurs
Membres identifiés du Cancéropôle Est :
Dr FRISCH Benoit
,
Dr HEURTAULT Béatrice
,
Dr KICHLER Antoine
Tous les auteurs :
Spanedda MV, De Giorgi M, Heurtault B, Kichler A, Bourel-Bonnet L, Frisch B
Lien Pubmed
Résumé
Click chemistry, and particularly azide-alkyne cycloaddition, represents one of the principal bioconjugation strategies that can be used to conveniently attach various ligands to the surface of preformed liposomes. This efficient and chemoselective reaction involves a Cu(I)-catalyzed azide-alkyne cycloaddition which can be performed under mild experimental conditions in aqueous media. Here we describe the application of a model click reaction to the conjugation, in a single step, of unprotected α-1-thiomannosyl ligands, functionalized with an azide group, to liposomes containing a terminal alkyne-functionalized lipid anchor. Excellent coupling yields were obtained in the presence of bathophenanthrolinedisulphonate, a water-soluble copper-ion chelator, acting as catalyst. No vesicle leakage was triggered by this conjugation reaction, and the coupled mannose ligands were exposed at the surface of the liposomes. The major limitation of Cu(I)-catalyzed click reactions is that this type of conjugation is restricted to liposomes made of saturated (phospho)lipids. To circumvent this constraint, an example of alternate copper-free azide-alkyne click reaction has been developed, and it was applied to the anchoring of a biotin moiety that was fully functional and could be therefore quantified. Molecular tools and results are presented here.
Mots clés
Azide–alkyne cycloaddition, Bioconjugation, Click chemistry, Copper-free click chemistry, Liposomes, Mannose
Référence
Methods Mol Biol. 2023 ;2622:173-189
