Institute of Electro-Optical Science and Technology, National Taiwan Normal University, No. 88, Section 4, Ting-Chou Road, Taipei 11677, Taiwan.
Institute of Electro-Optical Science and Technology, National Taiwan Normal University, No. 88, Section 4, Ting-Chou Road, Taipei 11677, Taiwan.
Biosens Bioelectron. 2017 Aug 15;94:351-357. doi: 10.1016/j.bios.2017.03.008. Epub 2017 Mar 7.
Specific peptide aptamers can be used in place of expensive antibody proteins, and they are gaining increasing importance as sensing probes due to their potential in the development of non-immunological assays with high sensitivity, affinity and specificity for human chorionic gonadotropin (hCG) protein. We combined graphene oxide (GO) sheets with a specific peptide aptamer to create a novel, simple and label-free tool to detect abnormalities at an early stage of pregnancy, a GO-peptide-based surface plasmon resonance (SPR) biosensor. This is the first binding interface experiment to successfully demonstrate binding specificity in kinetic analysis biomechanics in peptide aptamers and GO sheets. In addition to the improved affinity offered by the high compatibility with the target hCG protein, the major advantage of GO-peptide-based SPR sensors was their reduced nonspecific adsorption and enhanced sensitivity. The calculation of total electric field intensity (ΔE) in the GO-based sensing interfaces was significantly enhanced by up to 1.2 times that of a conventional SPR chip. The GO-peptide-based chip (1mM) had a high affinity (K) of 6.37×10M, limit of detection of 0.065nM and ultra-high sensitivity of 16 times that of a conventional SPR chip. The sensitivity of the slope ratio of the low concentration hCG protein assay in linear regression analysis was GO-peptide (1mM): GO-peptide (0.1mM): conventional chip (8-mercaptooctanoic acid)-peptide (0.1mM)=8.6: 3.3: 1. In summary, the excellent binding affinity, low detection limit, high sensitivity, good stability and specificity suggest the potential of this GO-peptide-based SPR chip detection method in clinical application. The development of real-time whole blood analytic and diagnostic tools to detect abnormalities at an early stage of pregnancy is a promising technique for future clinical application.
特定的肽适体可以替代昂贵的抗体蛋白，由于它们在开发具有高灵敏度、高亲和力和特异性的非免疫学检测方法方面的潜力，对于人绒毛膜促性腺激素 (hCG) 蛋白，它们作为传感探针的重要性日益增加。我们将氧化石墨烯 (GO) 片与特定的肽适体结合，创造了一种新型、简单且无需标记的工具，用于检测妊娠早期的异常情况，即基于 GO-肽的表面等离子体共振 (SPR) 生物传感器。这是第一个成功证明在肽适体和 GO 片的动力学分析生物力学中结合特异性的结合界面实验。除了与目标 hCG 蛋白的高兼容性提供的改进亲和力外，基于 GO-肽的 SPR 传感器的主要优势还在于其减少了非特异性吸附并提高了灵敏度。基于 GO 的传感界面中的总电场强度 (ΔE) 的计算值显著提高了 1.2 倍，与传统的 SPR 芯片相比。基于 GO-肽的芯片（1mM）具有高亲和力（K）为 6.37×10M、检测限为 0.065nM 和超高灵敏度，是传统 SPR 芯片的 16 倍。在线性回归分析中，低浓度 hCG 蛋白测定的斜率比灵敏度为 GO-肽（1mM）：GO-肽（0.1mM）：传统芯片（8-巯基辛酸-肽（0.1mM）=8.6：3.3：1。综上所述，优异的结合亲和力、低检测限、高灵敏度、良好的稳定性和特异性表明了基于 GO-肽的 SPR 芯片检测方法在临床应用中的潜力。开发实时全血分析和诊断工具以检测妊娠早期的异常情况是未来临床应用的有前途的技术。
