Versatile and efficient synthesis of protein–polysaccharide conjugate vaccines using aminooxy reagents and oxime chemistry

Applications of oxime chemistry are described for the efficient bioconjugation of proteins and polysaccharides for the preparation of conjugate vaccines. A number of approaches are described in this manuscript to functionalize proteins and polysaccharides with aminooxy (AO) groups and aldehydes which could then be covalently linked to each other via oxime formation, without the need for reduction. By using limiting numbers of active groups on each component, the extent of inter- and intramolecular crosslinking could be controlled. The approaches described are compatible and complementary to a number of chemistries currently used in conjugate vaccine synthesis. Oxime chemistry can be used to both simplify the synthesis of and increase yields of conjugate vaccines. Mice immunized with pneumococcal type 14 conjugates that were made using oxime chemistry mounted significant anti-polysaccharide immune responses. The primary immune response could be boosted, indicating that the polysaccharide conjugate had characteristics of a T cell dependent antigen.