A high affinity modified DNA aptamer containing base-appended bases for human β-defensin
A high affinity modified DNA aptamer containing base-appended bases for human β-defensin
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DOI：
10.1016/j.ab.2020.113627
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发表时间：
2020-04-01
影响因子：
2.9
通讯作者：
Waga, Iwao
中科院分区：
文献类型：
--
作者：
Minagawa, Hirotaka;Kataoka, Yuka;Waga, Iwao
我们使用碱基附加碱基修饰来开发一种新的腺嘌呤类似物，其在腺嘌呤的7位并入腺嘌呤衍生物。使用指数富集法与包括该类似物的修改的DNA文库的配体的系统进化，我们获得了A（ad 1），一种与唾液中发现的物理应激的生物标志物人β-防御素2强烈结合的适体。A（ad 1）相对于人β-防御素2的解离常数被发现是低的（6.8 nM），并被发现特异性地结合到唾液中的人β-防御素2与蛋白质，如使用磁珠下拉确认。据我们所知，没有先前的报告核酸适体特异性结合人β-防御素2。然而，我们的研究结果表明，这样的腺嘌呤类似物含有DNA文库是非常有效的收购高亲和力的适体。
We used base-appended base modification to develop a new adenine analog, which incorporates an adenine derivative at position 7 of adenine. Using the systematic evolution of ligands by exponential enrichment method with a modified DNA library including this analog, we obtained A(ad1), an aptamer that binds strongly to human beta-defensin 2, a biomarker of physical stress found in saliva. The dissociation constant of A(ad1) with respect to human beta-defensin 2 was found to be low (6.8 nM), and was found to bind specifically to human beta-defensin 2 in saliva spiked with the protein, as confirmed using pull-down with magnetic beads. To our knowledge, there are no prior reports of nucleic-acid aptamers that bind specifically to human beta-defensin 2. However, our results indicated that such adenine analog-containing DNA libraries are extremely effective in the acquisition of high-affinity aptamers.