Matsumoto I, Seno N, Golovtchenko-Matsumoto A M, Osawa T
J Biochem. 1980 Feb;87(2):535-40. doi: 10.1093/oxfordjournals.jbchem.a132775.
We have found a simple procedure to convert epoxy-activated agarose into amino derivatives using ammonia solution. The amino derivatives of agarose were succinylated with succinic anhydride and then activated with N-hydroxysuccinimide according to the method of Cuatrecasas and Parikh. Formula: (See Text). Coupling of the ligand to the activated agarose (shown above) was performed under mild conditions. The adsorbent thus obtained has no charged group in the linkage region between the ligand and agarose, thus reducing the nonspecific adsorption, and the bonds formed (ether and amide bonds) are stable even in an alkaline medium. Lens culinaris hemagglutinin-Sepharose 4B prepared by this method was successfully used for the affinity chromatography of solubilized human red blood cell membrane in detergent solution and was stable when elution was performed with borate buffer, pH 9.8.
我们发现了一种简单的方法，可使用氨溶液将环氧活化的琼脂糖转化为氨基衍生物。按照夸特雷卡斯和帕里克的方法，琼脂糖的氨基衍生物用琥珀酸酐进行琥珀酰化，然后用N-羟基琥珀酰亚胺活化。化学式：（见正文）。配体与上述活化琼脂糖的偶联在温和条件下进行。由此获得的吸附剂在配体与琼脂糖之间的连接区域没有带电基团，从而减少了非特异性吸附，并且形成的键（醚键和酰胺键）即使在碱性介质中也很稳定。用这种方法制备的菜豆凝集素-琼脂糖4B成功用于去污剂溶液中溶解的人红细胞膜的亲和层析，并用pH 9.8的硼酸盐缓冲液洗脱时很稳定。
