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作 者:吴璞强[1] 金雄华[1] 夏海锋[1] 郑志永[1] 饶志明[1]
机构地区:[1]江南大学工业生物技术教育部重点实验室,江苏无锡214122
出 处:《过程工程学报》2011年第4期666-670,共5页The Chinese Journal of Process Engineering
基 金:国家自然科学基金资助项目(编号:30970056);中央高校基本科研业务费专项资金资助项目(编号:JUSRP11012)
摘 要:采用烯丙基缩水甘油醚对国产交联琼脂糖凝胶进行活化,与巯基乙酸反应后,偶联N-羟基丁二酰亚胺(NHS),得到连接臂长度为10个原子、具有对氨基高度特异性的琼脂糖凝胶活化中间体.通过控制琼脂糖凝胶活化双键的量,可使NHS的修饰密度处在较宽范围(20~150μmol/mL),有效减少介质上杂基团的引入并有利于配基密度的设计.活化过程对介质的理化性能无明显影响,活化介质在30min内即可完成对L-苯丙氨酸的偶联,效率在90%以上,且在多种溶液中贮藏30d后配基稳定性良好,几乎无泄露.The homemade crosslinked agrose gel was first activated by allyl glycidyl ether, then reacted with mercaptoacetic acid and coupled with N-hydroxysuccinimide (NHS). Thus a kind of activated intermediate agarose gel with a 10-atoms-long arm and high specificity to amino-group was obtained. The modification density of NHS can be situated in a wide range of 20-150 μmol/mL by controlling the olefinic bond on the gel. This can reduce miscellaneous group coupling to the gel, and benefit for the ligand density design. The activated gel does not show any difference compared with original gel in physicochemical performance. The activated gel is easy to couple with L-phenylalanine in 30 min, and the coupling efficiency is greater than 90%. Moreover, the coupled ligands have been stable and without leaking during storage for 30 d.
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