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电化学多肽传感器用于检测登革热生物标志物 NS1。

An electrochemical peptide sensor for detection of dengue fever biomarker NS1.

机构信息

Department of Pharmaceutical Engineering, Daegu Haany University, Gyeongsan, 38610, Republic of Korea.

Department of Chemistry, Institute of Interdisciplinary Convergence Research, Research Institute of Halal Industrialization Technology, Chung-Ang University, 84 Heukseok-ro, Dongjak-gu, Seoul, 06974, Republic of Korea.

出版信息

Anal Chim Acta. 2018 Oct 5;1026:109-116. doi: 10.1016/j.aca.2018.04.005. Epub 2018 Apr 17.

Abstract

Dengue virus type 2 NS1 (DENV2 NS1) is a specific and sensitive protein biomarker for dengue fever diagnosis. In this study we used polyvalent phage display to identify unique affinity peptides that can bind NS1 protein. Cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS) were used to investigate the binding interactions. The potential affinity peptide-displayed phage from these methods was selected; its sequence was EHDRMHAYYLTR (R3#10). Amino acid sequence analysis showed that the peptide was rich in basic residues (two His and Arg). Among all the peptides tested, R3#10 showed the greatest decrease in current in CV and increase in impedance in EIS upon binding to NS1 proteins. EIS revealed that R3#10 phage clones were more specific towards NS1 proteins, as compared to bovine serum albumin or the M13 wild type used as control. Detection of NS1 proteins is in accordance with the electron-transfer resistance (R) value of the sensor layer, which is confirmed by EIS, and the K value of the R3#10 peptide while binding to the phage particles was measured. To the best of our knowledge, this is the first example of identification and characterization of NS1 binding affinity peptides using phage display technology and electrochemical methods. We concluded that these new peptide-displayed phages or free peptides from phages may have potential applications in dengue diagnosis.

摘要

登革热病毒 2 型 NS1(DENV2 NS1)是一种用于登革热诊断的特异性和敏感的蛋白质生物标志物。在这项研究中,我们使用多价噬菌体展示技术来识别可以与 NS1 蛋白结合的独特亲和肽。我们使用循环伏安法(CV)和电化学阻抗谱(EIS)来研究结合相互作用。从这些方法中选择潜在的亲和肽展示噬菌体;其序列为 EHDRMHAYYLTR(R3#10)。氨基酸序列分析表明,该肽富含碱性残基(两个 His 和 Arg)。在所有测试的肽中,R3#10 在与 NS1 蛋白结合时,CV 中的电流下降最大,EIS 中的阻抗增加最大。EIS 显示,与作为对照的牛血清白蛋白或 M13 野生型相比,R3#10 噬菌体克隆对 NS1 蛋白具有更高的特异性。NS1 蛋白的检测符合传感器层的电子转移电阻(R)值,这一点通过 EIS 得到了证实,并且测量了 R3#10 肽与噬菌体颗粒结合时的 K 值。据我们所知,这是首次使用噬菌体展示技术和电化学方法鉴定和表征 NS1 结合亲和肽。我们得出结论,这些新的肽展示噬菌体或来自噬菌体的游离肽可能在登革热诊断中有潜在的应用。

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