Abstract
Conjugation of synthetic oligosaccharides and native polysaccharides to proteins is an important tool in glycobiology to create vaccines and antigens to screen lectins, toxins, and antibodies. A novel approach to potentiate and profile the immune response to vaccines involves targeting antigens directly to dendritic cells (DCs), the key cells engaged in the immunization process. Inclusion of a carbohydrate ligand recognized by C-type lectins expressed on their cell surface ensures targeting of vaccines to DCs and improved immunological responses. Here we describe a strategy that permits three sequential orthogonal conjugation reactions to prepare glycoconjugates and apply them to the synthesis of a conjugate vaccine that is targeted for uptake by DCs. The carrier protein is treated with an azo-transfer reagent to convert accessible amino groups to azide and then amide bond formation via reaction with carboxylic acid side chains is used to attach amino tether groups of a ligand to the protein. Azide-alkyne Huisgen cycloaddition conjugation, “click chemistry” is used to attach a second ligand equipped with a propargyl group or an analogous terminal alkyne, and following reduction of protein azide groups back to amine, these amino acid side chains can be subjected to amide formation such as reaction with succinimide esters or homobifunctional coupling reagents such as dialkyl squarate.
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Acknowledgement
The research was made possible by grants awarded to D. R. B.; a Discovery grant from the Natural Science and Engineering Research Council of Canada and support from the Alberta Innovates Centers Program.
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Lipinski, T., Bundle, D.R. (2015). Temporary Conversion of Protein Amino Groups to Azides: A Synthetic Strategy for Glycoconjugate Vaccines. In: Lepenies, B. (eds) Carbohydrate-Based Vaccines. Methods in Molecular Biology, vol 1331. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-2874-3_9
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DOI: https://doi.org/10.1007/978-1-4939-2874-3_9
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-2873-6
Online ISBN: 978-1-4939-2874-3
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