title: Assay of multiplex proteins from cell metabolism based on tunable aptamer and microchip electrophoresis.
abstract: 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.
authors: 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.
authors: Xuexia
authors: X
authors: Lin
authors: Department of Chemistry, Beijing Key Laboratory of Microanalytical Methods and Instrumentation, Tsinghua University, Beijing 100084, China.
authors: Qiushui
authors: Q
authors: Chen
authors: Department of Chemistry, Beijing Key Laboratory of Microanalytical Methods and Instrumentation, Tsinghua University, Beijing 100084, China.
authors: Wu
authors: W
authors: Liu
authors: State Key Laboratory of Chemical Resource Engineering, Beijing University of Chemical Technology, Beijing 100029, China.
authors: Linglu
authors: L
authors: Yi
authors: Department of Chemistry, Beijing Key Laboratory of Microanalytical Methods and Instrumentation, Tsinghua University, Beijing 100084, China.
authors: Haifang
authors: H
authors: Li
authors: State Key Laboratory of Chemical Resource Engineering, Beijing University of Chemical Technology, Beijing 100029, China.
authors: Zhihua
authors: Z
authors: Wang
authors: Department of Chemistry, Beijing Key Laboratory of Microanalytical Methods and Instrumentation, Tsinghua University, Beijing 100084, China. Electronic address: jmlin@mail.tsinghua.edu.cn.
authors: Jin-Ming
authors: JM
authors: Lin
mesh_terms: D052157:Aptamers, Nucleotide / Q000737:chemistry* / Q000235:genetics; D000077214:Becaplermin; D015374:Biosensing Techniques; D046209:Electrophoresis, Microchip; D006046:Gold / Q000737:chemistry; D006801:Humans; D020574:Proto-Oncogene Proteins c-sis / Q000737:chemistry / Q000302:isolation & purification; D042461:Vascular Endothelial Growth Factor A / Q000737:chemistry / Q000302:isolation & purification
chemical_list: D052157:Aptamers, Nucleotide; D020574:Proto-Oncogene Proteins c-sis; C467484:VEGFA protein, human; D042461:Vascular Endothelial Growth Factor A; D000077214:Becaplermin; D006046:Gold
keywords: Aptamer; Microchip electrophoresis; PDGF-BB; Thrombin; VEGF(165)