Akanuma H, Yamasaki M
J Biochem. 1978 Dec;84(6):1357-62. doi: 10.1093/oxfordjournals.jbchem.a132257.
A novel method for the use of CM-Sephadex in affinity chromatography is described. This method involves a carbodiimide-mediated synthesis of a lactone derivative, which is subsequently hydrazinolyzed to CM-Sephadex hydrazide. More than 90% of the carboxyl groups was converted to the hydrazide form by this method without any discernible degradation of the Sephadex beads. The hydrazide beads were further converted to the axide form and coupled with D-arginine and D-phenylalanine derivatives. These Sephadex derivatives contained more than 0.5 mmol of each amino acid per g dry beads and were found to be effective and specific adsorbents for carboxypeptidase B. This CM-Sephadex hydrazide should be useful in affinity chromatography because the resulting specific adsorbents involve stable amide coupling linkages and retain the physical properties of their precursor, Sephadex, which are favorable for column operation.
本文描述了一种在亲和色谱中使用CM-葡聚糖凝胶的新方法。该方法涉及通过碳二亚胺介导合成内酯衍生物，随后将其肼解为CM-葡聚糖凝胶酰肼。通过该方法，超过90%的羧基被转化为酰肼形式，而葡聚糖凝胶珠没有任何明显降解。酰肼珠进一步转化为酰亚胺形式，并与D-精氨酸和D-苯丙氨酸衍生物偶联。这些葡聚糖凝胶衍生物每克干珠含有超过0.5 mmol的每种氨基酸，并且被发现是羧肽酶B的有效且特异性吸附剂。这种CM-葡聚糖凝胶酰肼在亲和色谱中应该是有用的，因为所得的特异性吸附剂涉及稳定的酰胺偶联键，并保留了其前体葡聚糖凝胶的物理性质，这有利于柱操作。
