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.
提供了一种方法,可以在中性条件下利用Diels-Alder环加成反应共价连接
碳水化合物、蛋白质、核酸和其他
生物分子。例如,活性碳-碳双键被连接到载体蛋白质的游离
氨基位点上,而共轭二烯被连接到
碳水化合物半抗原上。在非常温和的条件下,
碳水化合物和蛋白质组分自发耦合,提供含有每个载体蛋白质分子最多37个
碳水化合物半抗原单位的糖蛋白。该方法也适用于将
生物分子固定在凝胶或固体支持上。这些共轭产物可用作免疫原和分析和诊断试剂。