A method is provided for covalently linking carbohydrates, proteins,
nucleic acids, and other biomolecules under neutral conditions, using a
Diels-Alder cycloaddition reaction. In an example, activated
carbon-carbon double bonds were attached to free amino sites of a carrier
protein, and a conjugated diene was attached to a carbohydrate hapten.
Spontaneous coupling of the carbohydrate and the protein components under
very mild conditions provided glycoconjugates containing up to 37
carbohydrate hapten units per carrier protein molecule. The method is
also applicable to the immobilization of biomolecules on gel or solid
supports. The conjugated products are useful as immunogens and as
analytical and diagnostic reagents.