期刊
ANALYTICAL CHEMISTRY
卷 84, 期 14, 页码 6104-6109出版社
AMER CHEMICAL SOC
DOI: 10.1021/ac300997q
关键词
-
资金
- University of Utah
Here we report an aptamer-based analogue of the widely used sandwich enzyme-linked immunosorbent assay (ELISA). This assay utilizes the cocaine split aptamer, which is comprised of two DNA strands that only assemble in the presence of the target small molecule. One split aptamer fragment is immobilized on a microplate, then a test sample is added containing the second split aptamer fragment. If cocaine is present in the test sample, it directs assembly of the split aptamer and promotes a chemical ligation between azide and Cyclooctyne functional groups appended to the termini of the split aptamer fragments. Ligation results in covalent attachment of biotin to the microplate and provides a colorimetric output upon conjugation to streptavidin horseradish peroxidase. Using this assay, we demonstrate detection of cocaine at concentrations of 100 nM-100 mu M in buffer and 1-100 mu M human blood serum. The detection limit of 1 mu M in serum represents an improvement of two orders of magnitude over previously reported split aptamer-based sensors and highlights the utility of covalently trapping split aptamer assembly events.
作者
我是这篇论文的作者
点击您的名字以认领此论文并将其添加到您的个人资料中。
推荐
暂无数据