Anti-carbohydrate antibodies elicited by polyvalent display on a viral scaffold

Tetra- and hexasaccharides were arrayed on the exterior surface of cowpea mosaic virus by using a copper-catalyzed azidealkyne cycloaddition reaction. Inoculation of chickens with these virus conjugates gave rise to large quantities of polyclonal anti-glycan IgY antibodies that displayed excellent avidity and specificity on analysis with printed glycan microarrays. Avian IgY antibodies are produced in significantly higher yield than is possible for mouse or rabbit IgG, and exhibit reduced cross reactivity with native mammalian proteins. For a tri-LacNAc antigen, affinity purification against immobilized mono-LacNAc was necessary to provide a set of antibodies with specific binding properties. Comparable performance was observed for the virus-based polyclonal versus a commercial monoclonal antibody raised against the globo-H tetrasaccharide; this highlights the utility of the glycan microarray for both quality control and rapid in-depth analysis. Virus-carbohydrate conjugates ore promising candidates for development in diagnostic and immunotherapeutic applications.