Differentiation and Detection of PDGF Isomers and Their Receptors by Tunable Aptamer Capillary Electrophoresis

被引：48
作者：
Zhang, Hongquan [1 ]
Li, Xing-Fang [1 ]
Le, X. Chris [1 ]
机构：
[1] Univ Alberta, Div Analyt & Environm Toxicol, Dept Lab Med & Pathol, Fac Med & Dent, Edmonton, AB T6G 2G3, Canada
来源：
ANALYTICAL CHEMISTRY | 2009年 / 81卷 / 18期
基金：
加拿大健康研究院; 加拿大自然科学与工程研究理事会;
关键词：
GROWTH-FACTOR RECEPTORS; ROLLING CIRCLE AMPLIFICATION; SMOOTH-MUSCLE CELLS; ELECTROCHEMICAL DETECTION; GOLD NANOPARTICLES; PROTEIN; DNA; BINDING; ESTABLISHES; LIGANDS;
D O I：
10.1021/ac901471w
中图分类号：
O65 [分析化学];
学科分类号：
070302 ; 081704 ;
摘要：
Tunable aptamer capillary electrophoresis (CE) techniques were developed to enable the separation and detection of platelet derived growth factor (PDGF) isomers and their receptors. Using an aptamer that formed a stable complex with the B chain but not with the A chain of PDGF, we were able to tweak the electrophoretic mobilities of the PDGF isomers for their separation. PDGF-AB bound to a single aptamer molecule was well resolved from PDGF-BB bound to two aptamer molecules. Simultaneous determination of 50 pM of two isomers was accomplished in a single analysis, Further-more, PDGF-AB was used as a connector to bring receptor alpha and fluorescent aptamer into a single complex molecule. As a result, the formation of a (receptor alpha)-(PDGF-AB)-(fluorescent aptamer) ternary complex enabled the detection of the receptor a by tunable aptamer CE. A competitive assay was developed to determine receptor beta, making use of the competition between the receptor beta and fluorescent aptamer in binding to PDGF-BB. Detection limits were 0.5 nM for PDGF receptor alpha and 3 nM for receptor beta. Determination of PDGF isomers and their receptors in diluted serum samples showed no interference from the sample matrix.
引用
收藏
页码：7795 / 7800
页数：6
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