Diagnostic potential of PhotoSELEX-evolved ssDNA aptamers
Diagnostic potential of PhotoSELEX-evolved ssDNA aptamers
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DOI：
10.1016/s0168-1656(00)00290-x
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发表时间：
2000-08-25
影响因子：
4.1
通讯作者：
Koch, TH
中科院分区：
文献类型：
--
作者：
Golden, MC;Collins, BD;Koch, TH
证明了两种能够光交联重组人碱性成纤维细胞生长因子（bFGF（（115）的修饰的ssDNA适体的高灵敏度和特异性。通过一种新的，共价的，在体外选择方法，称为配体的光化学系统进化指数富集（PhotoSELEX）的适体进行了鉴定。适体表现出与市售ELISA单克隆抗体相当的对bFGF（（115））的高灵敏度，在主要测试条件下绝对灵敏度为至少0.058 ppt bFGF（（115））。这些适体将bFGF（115）与同源蛋白、血管内皮生长因子（VEGF）和血小板衍生生长因子（PDGF）精细地区分开。还描述了一种商业上可行的诊断系统，该系统包含能够同时定量大量分析物分子的PhotoSELEX进化的适体。这样的系统受益于适体与靶蛋白的共价键合，允许用变性剂剧烈洗涤以改善信噪比。(C)2000 Elsevier Science B.V.保留所有权利。
High sensitivity and specificity of two modified ssDNA aptamers capable of photocross-linking recombinant human basic fibroblast growth factor (bFGF((115))) were demonstrated. The aptamers were identified through a novel, covalent, in vitro selection methodology called photochemical systematic evolution of ligands by exponential enrichment (PhotoSELEX). The aptamers exhibited high sensitivity for bFGF((115)) comparable with commercially available ELISA monoclonal antibodies with an absolute sensitivity of at least 0.058 ppt bFGF((115)) under prevailing test conditions. The aptamers exquisitely distinguished bFGF((115)) from consanguine proteins, vascular endothelial growth factor (VEGF) and platelet derived growth factor (PDGF). A commercially viable diagnostic system incorporating PhotoSELEX-evolved aptamers capable of simultaneous quantification of a large number of analyte molecules is also described. Such a system benefits from covalent bonding of aptamer to target protein allowing vigorous washing with denaturants to improve signal to noise. (C) 2000 Elsevier Science B.V. All rights reserved.