Electrochemical aptameric recognition system for a sensitive protein assay based on specific target binding-induced rolling circle amplification.
Electrochemical aptameric recognition system for a sensitive protein assay based on specific target binding-induced rolling circle amplification.
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DOI：
10.1021/ac902400n
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发表时间：
2010-02
影响因子：
7.4
通讯作者：
Zai‐Sheng Wu;Hui Zhou;Songbai Zhang;G. Shen;R. Yu
中科院分区：
文献类型：
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作者：
Zai‐Sheng Wu;Hui Zhou;Songbai Zhang;G. Shen;R. Yu
提出了一种基于靶标结合诱导滚动圈扩增（RCA）的可重复使用的PDGF-BB蛋白核酸适配体识别系统。利用互补DNA （CDNA）、线性挂锁探针和引物探针将RCA过程引入到适体-靶标结合事件中，并通过CDNA的补体延长原适体，优雅地设计了新的适体。适配体传感系统促进了多种功能元件集成到一个信号方案中：独特的电化学技术，有吸引力的RCA过程，可逆的DNA杂交，以及理想的适配体目标识别。这种基于rca的电化学识别系统不仅表现出优异的性能（例如，检测限为6.3 x 10(-11) M，线性动态范围为2个数量级，高特异性和令人满意的重复性），而且还克服了传统的适体生物传感器相关的局限性（例如，信号靶结合依赖于特定的适体序列，或者每个目标分子需要两个或更多结合位点的三明治分析）。回收率试验证明了所开发的靶蛋白测定法的可行性。考虑到这些有吸引力的特性，这个适配体识别平台有望成为基础研究和应用研究中感兴趣的蛋白质和其他配体检测的候选平台。
A reusable aptameric recognition system was described for the electrochemical detection of the protein PDGF-BB based on the target binding-induced rolling circle amplification (RCA). A complementary DNA (CDNA), linear padlock probe, and primer probe were utilized to introduce a RCA process into the aptamer-target binding event while a new aptamer was elegantly designed via lengthening the original aptamer by the complement to the CDNA. The aptameric sensing system facilitates the integration of multiple functional elements into a signaling scheme: a unique electrochemical technique, an attractive RCA process, reversible DNA hybridization, and desirable aptameric target recognition. This RCA-based electrochemical recognition system not only exhibits excellent performance (e.g., a detection limit of 6.3 x 10(-11) M, a linear dynamic range of 2 orders of magnitude, high specificity, and satisfactory repeatability) but also overcomes the limitations associated with conventional aptameric biosensors (e.g., dependence of signaling target binding on specific aptamer sequence or requirement of sandwich assays for two or more binding sites per target molecule). A recovery test demonstrated the feasibility of the developed target protein assay. Given the attractive characteristics, this aptameric recognition platform is expected to be a candidate for the detection of proteins and other ligands of interest in both fundamental and applied research.