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Article Dans Une Revue Journal of visualized experiments : JoVE Année : 2020

Homogenous Glycoconjugate Produced by Combined Unnatural Amino Acid Incorporation and Click-Chemistry for Vaccine Purposes

Typhaine Violo
  • Fonction : Auteur
Christophe Dussouy
  • Fonction : Auteur
Charles Tellier
  • Fonction : Auteur
  • PersonId : 857372
Emilie Camberlein
  • Fonction : Auteur
  • PersonId : 1080952

Résumé

Genetic code expansion is a powerful tool to introduce unnatural amino acids (UAAs) into proteins to modify their characteristics, to study or create new protein functions or to have access to protein conjugates. Stop codon suppression, in particular amber codon suppression, has emerged as the most popular method to genetically introduce UAAs at defined positions. This methodology is herein applied to the preparation of a carrier protein containing an UAA harboring a bioorthogonal functional group. This reactive handle can next be used to specifically and efficiently graft a synthetic oligosaccharide hapten to provide a homogeneous glycoconjugate vaccine. The protocol is limited to the synthesis of glycoconjugates in a 1:1 carbohydrate hapten/carrier protein ratio but amenable to numerous pairs of biorthogonal functional groups. Glycococonjugate vaccine homogeneity is an important criterion to ensure complete physico-chemical characterization, thereby, satisfying more and more demanding drug regulatory agency recommendations, a criterion which is unmet by classical conjugation strategies. Moreover, this protocol makes it possible to finely tune the structure of the actual conjugate vaccine, giving rise to tools to address structure-immunogenicity relationships.
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Dates et versions

hal-02990572 , version 1 (05-11-2020)

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Typhaine Violo, Christophe Dussouy, Charles Tellier, Cyrille Grandjean, Emilie Camberlein. Homogenous Glycoconjugate Produced by Combined Unnatural Amino Acid Incorporation and Click-Chemistry for Vaccine Purposes. Journal of visualized experiments : JoVE, In press, ⟨10.3791/60821⟩. ⟨hal-02990572⟩
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