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改良丝网印刷电极用于开发高灵敏度无标记阻抗免疫传感器以检测淀粉样β肽。

Modified screen printed electrode for development of a highly sensitive label-free impedimetric immunosensor to detect amyloid beta peptides.

机构信息

School of Materials Science, Japan Advanced Institute of Science and Technology (JAIST), 1-1 Asahidai, Nomi, Ishikawa, 923-1292, Japan; School of Engineering Physics, Hanoi University of Science and Technology (HUST), No.1 Dai Co Viet, Hai Ba Trung, Hanoi, Viet Nam.

School of Materials Science, Japan Advanced Institute of Science and Technology (JAIST), 1-1 Asahidai, Nomi, Ishikawa, 923-1292, Japan.

出版信息

Anal Chim Acta. 2015 Sep 10;892:69-76. doi: 10.1016/j.aca.2015.08.036. Epub 2015 Aug 28.

DOI:10.1016/j.aca.2015.08.036
PMID:26388476
Abstract

Alzheimer's disease (AD) is a fatal neurodegenerative disease affecting approximately 26 million people world-wide, and the number is increasing as life expectancy increases. Since the only reliable diagnosis for the pathology is histochemical post-mortem examination, there is a rather urgent need for reliable, sensitive and quick detection techniques. Amyloid beta, being one of the established and widely accepted biomarkers of AD is a target biomolecule. Herein, we present fabrication of a labelless impedimetric amyloid beta immunosensor on carbon DEP (disposable electrochemical printed) chip. Three types of amyloid β impedimetric immunosensors were fabricated in a systematic step-wise manner in order to understand the effects that each surface modification chemistry had on detection sensitivity. We found that compared to a bare electrode, surface modification through formation of SAM of AuNPs increased sensitivity by approximately three orders of magnitude (LoD from 2.04 μM to 2.65 nM). A further modification using protein G, which helps orientate antibodies to an optimum position for interaction with antigen, lowered the LoD further to 0.57 nM. We have demonstrated that the presence of one of the most abundance proteins in biological fluids, bovine serum albumin (BSA), did not interfere with the sensitivity of the sensor. Since the DEP chips are disposable and the detection platform label-free, the developed sensor is relatively fast and cheap. These methods could easily be applied for detection of other antigens, with selection of the detection platform based on the desired for sensitivity.

摘要

阿尔茨海默病(AD)是一种致命的神经退行性疾病,影响着全球约 2600 万人,而且随着预期寿命的延长,这个数字还在不断增加。由于病理诊断的唯一可靠方法是组织化学死后检查,因此非常需要可靠、敏感和快速的检测技术。淀粉样β蛋白作为广泛接受的 AD 生物标志物之一,是一种目标生物分子。在此,我们展示了在碳 DEP(一次性电化学印刷)芯片上制备无标记的淀粉样β免疫传感器。我们以系统的逐步方式制备了三种类型的淀粉样β免疫传感器,以了解每种表面修饰化学对检测灵敏度的影响。我们发现,与裸电极相比,通过形成 AuNPs 的 SAM 进行表面修饰可将灵敏度提高约三个数量级(LoD 从 2.04 μM 降低至 2.65 nM)。进一步使用有助于将抗体定向到与抗原相互作用的最佳位置的蛋白 G 进行修饰,将 LoD 进一步降低至 0.57 nM。我们已经证明,在生物流体中含量最丰富的蛋白质之一牛血清白蛋白(BSA)的存在不会干扰传感器的灵敏度。由于 DEP 芯片是一次性的,并且检测平台是无标记的,因此开发的传感器相对快速且廉价。这些方法可以轻松应用于其他抗原的检测,根据所需的灵敏度选择检测平台。

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