利用金纳米粒子抗体共轭物通过比色免疫吸附法检测霉菌毒素。

IF 4.9 3区 工程技术 Q1 CHEMISTRY, ANALYTICAL Biosensors-Basel Pub Date : 2024-10-10 DOI:10.3390/bios14100491
Vinayak Sharma, Bilal Javed, Hugh J Byrne, Furong Tian
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引用次数: 0

摘要

金纳米粒子(AuNPs)具有优异的光学特性、强大的化学稳定性和易于生物共轭的特点,因此已成为科学研究中检测和生物传感应用的首选材料。本研究利用 AuNP 抗体(AuNP-mAb)共轭物开发了一种简单、快速、经济高效的比色免疫传感探针,用于检测黄曲霉毒素 B1 和玉米赤霉烯酮。抗毒素抗体通过物理吸附法附着在 AuNPs 上。所开发的比色免疫传感器的工作原理是,AuNP 的光学特性对聚集非常敏感,而临界高浓度盐可诱发聚集。虽然 AuNP 表面抗体的存在会抑制聚集,但这些抗体会以更高的亲和力与毒素结合,从而导致 AuNPs 表面暴露并在盐环境中聚集。由于 AuNPs 的表面等离子体共振频带发生了红色偏移,因此聚集会引发颜色从红色到紫色和蓝灰色的明显变化,但这种变化是可变的。转变的程度取决于毒素的暴露剂量,并可通过紫外-可见-近红外光谱使用校准曲线进行量化。经测定,玉米赤霉烯酮和黄曲霉毒素 B1 的检测限均低至 0.15 纳克/毫升。分析了制备的免疫探针在有其他霉菌毒素存在时对特定霉菌毒素的特异性。使用人工添加的样品对所开发的免疫探针的实际适用性进行了评估。与其他免疫测定方法相比,这种基于局部表面等离子体共振(LSPR)的比色免疫探针具有检测限低、读取速度快、成本低和易于制造等优点。
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Mycotoxin Detection through Colorimetric Immunoprobing with Gold Nanoparticle Antibody Conjugates.

Driven by their exceptional optical characteristics, robust chemical stability, and facile bioconjugation, gold nanoparticles (AuNPs) have emerged as a preferred material for detection and biosensing applications in scientific research. This study involves the development of a simple, rapid, and cost-effective colorimetric immuno-sensing probe to detect aflatoxin B1 and zearalenone using AuNP antibody (AuNP-mAb) conjugates. Anti-toxin antibodies were attached to the AuNPs by using the physical adsorption method. The colorimetric immunosensor developed operates on the principle that the optical properties of the AuNP are very sensitive to aggregation, which can be induced by a critical high salt concentration. Although the presence of antibodies on the AuNP surface inhibits the aggregation, these antibodies bind to the toxin with higher affinity, which leads to exposure of the surface of AuNPs and aggregation in a salt environment. The aggregation triggers a noticeable but variable alteration in color from red to purple and blueish gray, as a result of a red shift in the surface plasmon resonance band of the AuNPs. The extent of the shift is dependent on the toxin exposure dose and can be quantified using a calibration curve through UV-Visible-NIR spectroscopy. The limit of detection using this assay was determined to be as low as 0.15 ng/mL for both zearalenone and aflatoxin B1. The specificity of the prepared immunoprobe was analyzed for a particular mycotoxin in the presence of other mycotoxins. The developed immunoprobe was evaluated for real-world applicability using artificially spiked samples. This colorimetric immunoprobe based on localized surface plasmon resonance (LSPR) has a reduced detection limit compared to other immunoassays, a rapid readout, low cost, and facile fabrication.

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来源期刊
Biosensors-Basel
Biosensors-Basel Biochemistry, Genetics and Molecular Biology-Clinical Biochemistry
CiteScore
6.60
自引率
14.80%
发文量
983
审稿时长
11 weeks
期刊介绍: Biosensors (ISSN 2079-6374) provides an advanced forum for studies related to the science and technology of biosensors and biosensing. It publishes original research papers, comprehensive reviews and communications. Our aim is to encourage scientists to publish their experimental and theoretical results in as much detail as possible. There is no restriction on the length of the papers. The full experimental details must be provided so that the results can be reproduced. Electronic files and software regarding the full details of the calculation or experimental procedure, if unable to be published in a normal way, can be deposited as supplementary electronic material.
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