Department of Chemistry and Chemical Engineering, Hunan University of Arts and Science, Changde 415000, PR China; State Key Laboratory for Chemo/Biosensing and Chemometrics, College of Chemistry and Chemical Engineering, Hunan University, Changsha 410082, PR China.
Department of Chemistry and Chemical Engineering, Hunan University of Arts and Science, Changde 415000, PR China.
Biosens Bioelectron. 2015 Apr 15;66:363-9. doi: 10.1016/j.bios.2014.11.044. Epub 2014 Nov 24.
Using platelet-derived growth factor B chain dimer (PDGF-BB) as the model target, a background current eliminated electrochemical aptameric sensing platform for highly sensitive and signal-on detection of protein is proposed in this paper. Successful fabrication of the biosensor depends on ingenious design of aptamer probe, which contains the aptamer sequence for PDGF-BB and the recognition sequence for EcoRI endonuclease. In the absence of PDGF-BB, the ferrocene labeled aptamer probe folds into a hairpin structure and forms a recognition site for EcoRI. By treatment with endonuclease, the specific and cleavable double-stranded region is cut off and redox-active ferrocene molecule is removed from the electrode surface, and almost no peak current is observed. When binding with target protein, the designed aptamer probe changes its conformation and dissociates the recognition double strand. The integrated aptamer probe is maintained when exposing to EcoRI endonuclease, resulting in obvious peak current. Therefore, a signal-on and sensitive sensing strategy for PDGF-BB detection is fabricated with eliminated background current. Under the optimized experimental conditions, a wide linear response range of 4 orders of magnitude from 20pgmL(-1) to 200ngmL(-1) is achieved with a detection limit of 10pgmL(-1). Moreover, the present aptameric platform is universal for the analysis of a broad range of target molecules of interest by changing and designing the sequence of aptamer probe.
本文提出了一种基于血小板衍生生长因子 B 链二聚体(PDGF-BB)的背景电流消除电化学适体传感平台,用于高灵敏度和信号开启的蛋白质检测。生物传感器的成功制备依赖于适体探针的巧妙设计,其中包含 PDGF-BB 的适体序列和 EcoRI 内切酶的识别序列。在没有 PDGF-BB 的情况下,标记了电化学活性物质的适体探针折叠成发夹结构,并形成 EcoRI 的识别位点。通过内切酶处理,特异性和可切割的双链区域被切断,并且电化学活性物质从电极表面被去除,几乎观察不到峰电流。当与靶蛋白结合时,设计的适体探针改变其构象并解离识别双链。当暴露于 EcoRI 内切酶时,整合的适体探针得以保持,从而导致明显的峰电流。因此,通过消除背景电流,构建了一种用于 PDGF-BB 检测的信号开启和灵敏传感策略。在优化的实验条件下,实现了从 20pgmL(-1)到 200ngmL(-1)的 4 个数量级的宽线性响应范围,检测限为 10pgmL(-1)。此外,通过改变和设计适体探针的序列,本适体平台可用于广泛的目标分子分析,具有通用性。