An Aptamer-Based Bound/Free Separation System for Protein Detection
An Aptamer-Based Bound/Free Separation System for Protein Detection
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DOI：
10.1002/elan.200804555
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发表时间：
2009-06-01
期刊：
影响因子：
3
通讯作者：
Ikebukuro, Kazunori
中科院分区：
文献类型：
--
作者：
Fukasawa, Mieko;Yoshida, Wataru;Ikebukuro, Kazunori
适体与其互补链杂交。然而，互补链难以与结合至靶标的适体杂交，因为适体形成更高级的结构。利用这一特性，我们开发了简单的结合/游离分离系统的凝血酶和IgE检测。将互补链固定在珠上，并用吡咯喹啉醌葡萄糖脱氢酶（PQQGDH）标记适体。在不存在靶标的情况下，适体被珠捕获，而在存在靶标的情况下。与靶结合的适体不被捕获。因此，可以容易地回收适体-靶复合物并通过PQQGDH活性检测。该系统允许检测270 pM凝血酶和1 nM IgE。
Aptamer hybridizes with its complementary strand. However, the complementary strand has difficulties to hybridize with the aptamer bound to a target because the aptamer forms higher-order structures. Exploiting this property, we developed simple bound/free separation systems for thrombin and IgE detection. The complementary strand was immobilized onto beads and the aptamer was labeled with pyrroquinoline quinone glucose dehydrogenase (PQQGDH). In the absence of a target, the aptamer is trapped by beads, whereas in the presence of a target. the aptamer bound to the target is not trapped. Thus the aptamer-target complexes can be recovered easily and detected by PQQGDH activity. This system allow the detection of 270 pM thrombin and 1 nM IgE.