Department of Chemical Science and Engineering, Tokyo Institute of Technology, 2-12-1 O-okayama, Meguro-ku, Tokyo, 152-8552, Japan.
School of Physics and Astronomy, University of Leeds, Leeds, LS2 9JT, United Kingdom.
Biosens Bioelectron. 2020 Apr 1;153:112030. doi: 10.1016/j.bios.2020.112030. Epub 2020 Jan 20.
A novel peptide-based three-dimensional probe called "peptide matrix," inspired by the antibody paratope region, was fabricated on a surface plasmon resonance (SPR) sensor chip to enhance the sensitivity of detecting the explosive 2,4,6-trinitrotoluene (TNT). Although peptide aptamer is an attractive candidate for a molecular recognition probe because of its ease of synthesis and chemical stability, it still has difficulty in applying to highly sensitive (i.e. parts-per-billion (ppb) or sub-ppb level) detections. Thus, we developed the concept of peptide matrix structure, which is constructed by consecutive disulfide bond formation between a large number of peptide fragments. This robust three-dimensional structure displays multiple binding sites which can efficiently associate with each TNT molecule. The peptide matrix lowered the dissociation constant (K) by two orders of magnitude compared to the linear peptide aptamer, estimating K as 10.1 nM, which is the lowest concentration reported by using peptide-based TNT probe. Furthermore, the concentration limit of detection of peptide matrix modified SPR sensor was 0.62 ppb, and hence comparable to single-chain variable fragment (scFv)-based TNT sensors. To our knowledge, this is the first report demonstrating peptide matrix fabrication and its application for small explosive molecule detection. This peptide matrix-based approach, which has the advantage of simple synthesis and high sensitivity, will be applicable to many other small-molecule label-free detections.
一种新型的基于肽的三维探针,称为“肽基质”,灵感来自抗体的互补决定区,被制备在表面等离子体共振(SPR)传感器芯片上,以提高检测爆炸物 2,4,6-三硝基甲苯(TNT)的灵敏度。尽管肽适体由于其易于合成和化学稳定性,是一种有吸引力的分子识别探针候选物,但它在应用于高灵敏度(即十亿分之一(ppb)或亚 ppb 级)检测方面仍然存在困难。因此,我们开发了肽基质结构的概念,该结构是通过大量肽片段之间的连续二硫键形成构建的。这种坚固的三维结构显示出多个结合位点,可以有效地与每个 TNT 分子结合。与线性肽适体相比,肽基质将解离常数(K)降低了两个数量级,估计 K 值为 10.1 nM,这是使用基于肽的 TNT 探针报告的最低浓度。此外,肽基质修饰的 SPR 传感器的浓度检测限为 0.62 ppb,因此可与单链可变片段(scFv)-基于 TNT 传感器相媲美。据我们所知,这是第一个报告展示肽基质制备及其在小分子爆炸物检测中的应用的报告。这种基于肽基质的方法具有合成简单和灵敏度高的优点,将适用于许多其他小分子无标记检测。