High specificity, electrochemical sandwich assays based on single aptamer sequences and suitable for the direct detection of small-molecule targets in blood and other complex matrices.

High specificity, electrochemical sandwich assays based on single aptamer sequences and suitable for the direct detection of small-molecule targets in blood and other complex matrices.
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DOI:
10.1021/ja901315w
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发表时间:
2009-05-27
影响因子:
15
通讯作者:
Plaxco KW
Plaxco KW
中科院分区:
化学1区
文献类型:
--
作者:
Zuo X;Xiao Y;Plaxco KW

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我们在此证明了基于单个适体序列的夹心测定法适用于直接检测血清和其他复杂基质中的小分子靶标。通过将适体分成两部分,我们将单一亲和试剂转化为双组分系统,其中靶标的存在驱动由靶标和适体的两个半部组成的复合物的形成。为了证明这种方法的实用性,我们已经使用了单一的抗可卡因和抗ATP适体制造针对代表性小分子coaine和ATP的电化学传感器。这两种目标都是在低微摩尔浓度下,在几秒钟内,并在一个方便的,一般的,易于重复使用的电化学格式检测。此外,两种传感器都具有足够的选择性,可以直接部署在血液、粗细胞裂解物和其他复杂的样品基质中。
We herein demonstrate a sandwich assay based on single aptamer sequences is suitable for the direct detection of small molecule targets in blood serum and other complex matrices. By splitting an aptamer into two pieces, we convert a single affinity reagent into a two-component system in which the presence of the target drives formation of a complex comprised of the target and the two halves of the aptamer. To demonstrate the utility of this approach we have used single anticocaine and anti-ATP aptamers to fabricate electrochemical sensors directed against the representative small molecules coaine and ATP. Both targets are detected at low micromolar concentrations, in seconds, and in a convenient, general, readily reusable, electrochemical format. Moreover, both sensors are selective enough to deploy directly in blood, crude cellular lysates and other complex sample matrices.
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