State Key Laboratory of Chemical Resource Engineering, Beijing University of Chemical Technology, Beijing 100029, China; Department of Chemistry, Beijing Key Laboratory of Microanalytical Methods and Instrumentation, Tsinghua University, Beijing 100084, China.
Department of Chemistry, Beijing Key Laboratory of Microanalytical Methods and Instrumentation, Tsinghua University, Beijing 100084, China.
Biosens Bioelectron. 2015 Jan 15;63:105-111. doi: 10.1016/j.bios.2014.07.013. Epub 2014 Jul 11.
A simple and rapid method for multiplex protein assay based on tunable aptamer by microchip electrophoresis has been developed. Different lengths of aptamers can modulate the electrophoretic mobility of proteins, allowing the protein molecules to be effectively separated in hydroxyethyl cellulose buffer with 1.00 mM magnesium ion. A non-specific DNA was exploited as an internal standard to achieve the quantitative assay and to reduce the interference. A fluorescence dye SYBR gold was exploited to improve the sensitivity and to suppress the interference from sample matrix. Under optimum conditions, quantitative assay of PDGF-BB (R(2)=0.9986), VEGF165 (R(2)=0.9909), and thrombin (R(2)=0.9947) were achieved with a dynamic range in the 5.00-150.0 nM and RSDs in the 5.87-16.3% range. The recoveries were varied from 83.6% to 113.1%. Finally, the proposed method was successfully applied to analyze cell secretions, and then the concentration of PDGF-BB and VEGF165 were detected from 5.15 nM to 2.03 nM, and 3.14 to 2.53 nM, respectively, indicating the established method can be used to analyze cell secretions.
基于微流控芯片电泳的可调适适体的多重蛋白分析的快速简单方法已经建立。不同长度的适体可以调节蛋白质的电泳迁移率，使蛋白质分子在含有 1.00 mM 镁离子的羟乙基纤维素缓冲液中得到有效分离。非特异性 DNA 被用作内标以实现定量分析并减少干扰。利用荧光染料 SYBR Gold 提高了灵敏度并抑制了来自样品基质的干扰。在最佳条件下，实现了 PDGF-BB（R（2）=0.9986）、VEGF165（R（2）=0.9909）和凝血酶（R（2）=0.9947）的定量分析，动态范围为 5.00-150.0 nM，RSD 在 5.87-16.3%范围内。回收率在 83.6%-113.1%之间变化。最后，该方法成功地应用于分析细胞分泌物，并检测到 PDGF-BB 和 VEGF165 的浓度分别为 5.15 nM 至 2.03 nM 和 3.14 至 2.53 nM，表明所建立的方法可用于分析细胞分泌物。
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