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利用增强型 SPR 和 p(HEMA)刷敏感快速检测牛奶中的黄曲霉毒素 M1。

Sensitive and rapid detection of aflatoxin M1 in milk utilizing enhanced SPR and p(HEMA) brushes.

机构信息

Department of Medical Engineering and Biotechnology, Ernst-Abbe-University of Applied Sciences Jena, Carl-Zeiss Promenade 2, 07745 Jena, Germany; Institute of Analytical Chemistry, University of Regensburg, Josef-Engert Straße, 93053 Regensburg, Germany.

Department of Medical Engineering and Biotechnology, Ernst-Abbe-University of Applied Sciences Jena, Carl-Zeiss Promenade 2, 07745 Jena, Germany.

出版信息

Biosens Bioelectron. 2016 Jul 15;81:159-165. doi: 10.1016/j.bios.2016.02.061. Epub 2016 Feb 24.

Abstract

The rapid and sensitive detection of aflatoxin M1 (AFM1) in milk by using surface plasmon resonance (SPR) biosensor is reported. This low molecular weight mycotoxin is analyzed using an indirect competitive immunoassay that is amplified by secondary antibodies conjugated with Au nanoparticles. In order to prevent fouling on the sensor surface by the constituents present in analyzed milk samples, an interface with poly(2-hydroxyethyl methacrylate) p(HEMA) brush was employed. The study presents a comparison of performance characteristics of p(HEMA)-based sensor with a regularly used polyethylene glycol-based architecture relying on mixed thiol self-assembled monolayer. Both sensors are characterized in terms of surface mass density of immobilized AFM1 conjugate as well as affinity bound primary and secondary antibodies. The efficiency of the amplification strategy based on Au nanoparticle is discussed. The biosensor allowed for highly sensitive detection of AFM1 in milk with a limit of detection (LOD) as low as 18pgmL(-1) with the analysis time of 55min.

摘要

本文报道了一种利用表面等离子体共振(SPR)生物传感器快速灵敏检测牛奶中黄曲霉毒素 M1(AFM1)的方法。该低分子量真菌毒素采用间接竞争免疫分析法进行分析,该方法通过与金纳米粒子偶联的二级抗体进行放大。为了防止分析牛奶样品中存在的成分在传感器表面上发生污垢,使用了带有聚(2-羟乙基甲基丙烯酸酯)p(HEMA)刷的界面。本研究比较了基于 p(HEMA)的传感器与基于混合硫醇自组装单层的常规使用的聚乙二醇基结构的性能特征。两种传感器均在固定化 AFM1 缀合物的表面质量密度以及亲和结合的一级和二级抗体方面进行了表征。讨论了基于金纳米粒子的放大策略的效率。该生物传感器允许在牛奶中高度灵敏地检测 AFM1,检测限(LOD)低至 18pgmL(-1),分析时间为 55min。

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