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基于磁珠的比色免疫分析法检测黄曲霉毒素B1:使用金纳米颗粒

Magnetic bead-based colorimetric immunoassay for aflatoxin B1 using gold nanoparticles.

作者信息

Wang Xu, Niessner Reinhard, Knopp Dietmar

机构信息

Institute of Hydrochemistry and Chemical Balneology, Chair for Analytical Chemistry, Technische Universität München, Marchioninistrasse 17, Munich 81377, Germany.

出版信息

Sensors (Basel). 2014 Nov 14;14(11):21535-48. doi: 10.3390/s141121535.

Abstract

A competitive colorimetric immunoassay for the detection of aflatoxin B1 (AFB) has been established using biofunctionalized magnetic beads (MBs) and gold nanoparticles (GNPs). Aflatoxin B1-bovine serum albumin conjugates (AFB-BSA) modified MBs were employed as capture probe, which could specifically bind with GNP-labeled anti-AFB antibodies through immunoreaction, while such specific binding was competitively inhibited by the addition of AFB. After magnetic separation, the supernatant solution containing unbound GNPs was directly tested by UV-Vis spectroscopy. The absorption intensity was directly proportional to the AFB concentration. The influence of GNP size, incubation time and pH was investigated in detail. After optimization, the developed method could detect AFB in a linear range from 20 to 800 ng/L, with the limit of detection at 12 ng/L. The recoveries for spiked maize samples ranged from 92.8% to 122.0%. The proposed immunoassay provides a promising approach for simple, rapid, specific and cost-effective detection of toxins in the field of food safety.

摘要

利用生物功能化磁珠(MBs)和金纳米颗粒(GNPs)建立了一种用于检测黄曲霉毒素B1(AFB)的竞争性比色免疫分析方法。用黄曲霉毒素B1 - 牛血清白蛋白缀合物(AFB - BSA)修饰的磁珠作为捕获探针,其可通过免疫反应与GNP标记的抗AFB抗体特异性结合,而这种特异性结合会因加入AFB而受到竞争性抑制。磁分离后,含有未结合GNPs的上清液直接用紫外 - 可见光谱法检测。吸收强度与AFB浓度成正比。详细研究了GNP尺寸、孵育时间和pH的影响。经过优化,所建立的方法可在20至800 ng/L的线性范围内检测AFB,检测限为12 ng/L。加标玉米样品的回收率在92.8%至122.0%之间。所提出的免疫分析方法为食品安全领域中简单、快速、特异性和经济高效地检测毒素提供了一种有前景的方法。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7254/4279548/0a0b1583e7ab/sensors-14-21535f1.jpg

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