Aptamer-Linked Assay for Thrombin Using Gold - Mass Spectrometry Detection
Aptamer-Linked Assay for Thrombin Using Gold - Mass Spectrometry Detection
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DOI：
10.1021/ac900961y
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发表时间：
2009-09-01
影响因子：
7.4
通讯作者：
Le, X. Chris
中科院分区：
文献类型：
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作者：
Zhao, Qiang;Lu, Xiufen;Le, X. Chris
我们描述了一种灵敏和特异性的人α-凝血酶的夹心测定法。该检测利用两种亲和适体的夹心结合以提高特异性，利用金纳米颗粒进行信号放大，利用磁珠进行快速磁分离，利用电感耦合等离子体质谱进行超灵敏检测。其他蛋白质，如免疫球蛋白G、血清白蛋白、转铁蛋白、纤维蛋白原和溶菌酶未显示对人α-凝血酶测定的干扰。人α-凝血酶的检测限低至0.5 fmol，对应于50 μ L中的10 pM凝血酶，并且动态范围覆盖类似于3个数量级。
We describe a sensitive and specific sandwich assay for human alpha-thrombin. The assay takes advantage of sandwich binding of two affinity aptamers for increased specificity, gold nanoparticles for signal amplification, magnetic beads for fast magnetic separation, and inductively coupled plasma mass spectrometry for ultrasensitive detection. Other proteins, such as immunoglobulin G, serum albumin, transferrin, fibrinogen, and lysozyme did not show interference with the assay for human alpha-thrombin. The detection limit of human alpha-thrombin was as low as 0.5 fmol, corresponding to 10 pM thrombin in 50 mu L, and the dynamic range covered similar to 3 orders of magnitude.