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An electrochemical impedimetric sensing platform based on a peptide aptamer identified by high-throughput molecular docking for sensitive l-arginine detection.

基于高通量分子对接鉴定的肽适配子的电化学阻抗传感平台,用于灵敏的l-精氨酸检测。

  • 影响因子:4.37
  • DOI:10.1016/j.bioelechem.2020.107634
  • 作者列表:"He Y","Zhou L","Deng L","Feng Z","Cao Z","Yin Y
  • 发表时间:2021-02-01
Abstract

:As a primary building block for protein synthesis, l-arginine (l-Arg) is also a precursor for the synthesis of important metabolites, and is involved in various physiological and pathophysiological processes. l-Arg is a potential biomarker in clinical diagnosis and nutritional status assessment, making it valuable to quantify and monitor this biomolecule. In this study, peptide aptamers that specifically interact with l-Arg were identified by high-throughput molecular docking, and the binding capacities between the synthesized peptide aptamers and l-Arg were then measured by isothermal titration calorimetry. We hypothesized that the peptide aptamer with the greatest binding capacity could be used as the recognition element in a biosensor. A chemosynthetic peptide aptamer modified with mercaptan and spacer units (thioctic acid-GGGG-FGHIHEGY) was thus used to construct label-free electrochemical impedimetric biosensors for l-Arg based on gold electrodes. The optimum biosensor showed good sensitivity to l-Arg with a linear range of 0.1 pM-0.1 mM, and the calculated limit of detection (three times the signal-to-noise ratio) was 0.01 pM. Interference studies and assays of diluted serum samples were also carried out, and satisfactory results obtained. In conclusion, a potential method of peptide aptamer screening and biosensor fabrication for detecting small biological molecules was demonstrated.

摘要

: 作为蛋白质合成的主要组成部分,l-精氨酸 (l-Arg) 也是合成重要代谢物的前体,参与多种生理和病理生理过程。l-Arg是临床诊断和营养状态评估中的潜在生物标志物,使得定量和监测该生物分子具有价值。本研究通过高通量分子对接鉴定了与l-Arg特异性相互作用的肽适配体,然后通过等温滴定量热法测定了合成的肽适配体与l-Arg的结合能力。我们假设具有最大结合能力的肽适体可以用作生物传感器中的识别元件。因此,使用硫醇和间隔单元修饰的化学合成肽适体 (硫辛酸-GGGG-FGHIHEGY) 构建基于金电极的l-Arg的无标记电化学阻抗生物传感器。最佳生物传感器对l-Arg显示出良好的灵敏度,线性范围为0.1 ± pM-0.1 mM,计算的检测限 (信噪比的三倍) 为0.01 pM。还进行了稀释血清样品的干扰研究和测定,并获得了满意的结果。总之,证明了用于检测生物小分子的肽适体筛选和生物传感器制备的潜在方法。

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