Development of a High Sensitivity Rapid Sandwich ELISA Procedure and Its Comparison with the Conventional Approach
Development of a High Sensitivity Rapid Sandwich ELISA Procedure and Its Comparison with the Conventional Approach
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DOI：
10.1021/ac101339q
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发表时间：
2010-08-15
影响因子：
7.4
通讯作者：
O'Kennedy, Richard
中科院分区：
文献类型：
--
作者：
Dixit, Chandra Kumar;Vashist, Sandeep Kumar;O'Kennedy, Richard
建立了一种高灵敏度、快速的夹心酶联免疫吸附试验（ELISA）检测人胎球蛋白A/AHSG（α 2-HS糖蛋白）的方法。使用1-乙基-3 [3-二甲基氨基丙基]丁二酰亚胺盐酸盐和基于N-羟基磺基琥珀酰亚胺的异双功能交联将抗人胎球蛋白A抗体固定在氨基丙基三乙氧基硅烷化胺官能化微量滴定板上。开发的测定的分析灵敏度为39 pg/mL，而常规测定的分析灵敏度为625 pg/ mL。通过在不同的聚合物基质上进行人胎球蛋白A测定，即，聚苯乙烯、聚（甲基丙烯酸甲酯）和聚环烯烃（Zeonex），以改进的微量滴定板形式。因此，新开发的程序比现有的方法具有相当大的优势。
A highly sensitive and rapid sandwich enzyme-linked immunosorbent assay (ELISA) procedure was developed for the detection of human fetuin A/AHSG (alpha 2-HSglycoprotein), a specific biomarker for hepatocellular carcinoma and atherosclerosis. Anti-human fetuin A antibody was immobilized on aminopropyltriethoxysilanemediated amine-functionalized microtiter plates using 1-ethyl-3[3-dimethylaminopropylicarbodiimide hydrochloride and N-hydroxysulfosuccinimide-based heterobifunctional cross-linking. The analytical sensitivity of the developed assay was 39 pg/mL, compared to 625 pg/ mL for the conventional assay. The generic nature of the developed procedure was demonstrated by performing human fetuin A assays on different polymeric matrixes, i.e., polystyrene, poly(methyl methacrylate), and polycyclo-olefin (Zeonex), in a modified microliter plate format. Thus, the newly developed procedure has considerable advantages over the existing method.