Pepper D S
National Science Laboratory, Edinburgh, Scotland, UK.
Mol Biotechnol. 1994 Oct;2(2):157-78. doi: 10.1007/BF02824808.
Three different coupling chemistries that have been tried and tested for use in affinity chromatography are described. These methods are particularly recommended for use by workers who do not have access to, or do not wish to use, complex organic chemical synthetic procedures. They have been demonstrated repeatedly to be reliable, efficient, low cost, and easily scaleable up or down in size. The periodate oxidation method works best with Sephacryl type gels and uses only low toxicity reagents and couples well to proteins with both high efficiency and high capacity. The vinyl sulfone method is more reactive and couples both carbohydrates and proteins. The bis-epoxide method, although less reactive, can be used under more extreme conditions of pH to couple otherwise unreactive molecules, such as synthetic polymers, drugs, and so forth.
本文描述了三种已在亲和色谱中经过试验和测试的不同偶联化学方法。这些方法特别推荐给那些无法获得或不希望使用复杂有机化学合成程序的工作人员。它们已被反复证明是可靠、高效、低成本的,并且在规模大小上易于扩大或缩小。高碘酸盐氧化法与Sephacryl型凝胶配合效果最佳,仅使用低毒性试剂,并且能高效、高容量地与蛋白质偶联。乙烯砜法反应性更强,能与碳水化合物和蛋白质都进行偶联。双环氧化物法虽然反应性较弱,但可在更极端的pH条件下使用,以偶联其他不反应的分子,如合成聚合物、药物等。