Development of a Novel Aptamer-Antibody Sandwich Chemiluminescent Biosensor and Its Application in the Detection of Aflatoxin B₁.

IF 5.6 3区工程技术 Q1 CHEMISTRY, ANALYTICAL

Biosensors-Basel Pub Date : 2025-08-15 DOI:10.3390/bios15080538

Zhike Zhao, Jianghao Feng, Caizhang Wu

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Abstract

In addressing the challenges posed by high costs, low accuracy, and cumbersome operations in mycotoxin detection, a novel aptamer-antibody sandwich chemiluminescent biosensor for detecting aflatoxin B₁ (AFB₁) was developed. The indirect competition between AFB₁, aflatoxin B₁-ovomucoid complete antigen (AFB₁-OVA), and rabbit anti-ovomucoid (OVA) antibody results in the formation of a sandwich complex. This sandwich assay is linked to a horseradish peroxidase-labeled antibody, which catalyzes luminol chemiluminescence for the indirect detection of AFB₁. The biosensor was designed to operate with high precision, low cost, and a low detection limit for AFB₁, which is contingent upon experimental conditions such as pH, reagent concentration, temperature, and incubation time. The optimization of pH, aptamer concentration, competitive incubation time, competitive incubation temperature, and HRP-labeled antibody concentration was instrumental in achieving these objectives. Experimental findings demonstrated that the sensor's detection limit was 0.067 ng/mL, exhibiting excellent linearity (R² = 0.99679) within the concentration range of 0.25-10 ng/mL. The recovery rate of spiked samples ranged from 94.4% to 108.05%. This sensor boasts a low detection limit, straightforward operation, and minimal cost, thus offering a novel solution for developing cost-effective, high-precision mycotoxin detection methods.

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新型适配体-抗体夹心化学发光生物传感器的研制及其在黄曲霉毒素B1检测中的应用。

为解决霉菌毒素检测成本高、准确性低、操作繁琐等问题，研制了一种新型的适体-抗体夹心化学发光生物传感器，用于检测黄曲霉毒素B1 （AFB1）。AFB1、黄曲霉毒素b1 -卵细胞样完全抗原（AFB1-OVA）和兔抗卵细胞样抗体（OVA）之间的间接竞争导致三明治复合物的形成。这种夹心法与辣根过氧化物酶标记的抗体相关联，该抗体催化发光胺化学发光间接检测AFB1。该生物传感器设计具有高精度、低成本和AFB1低检出限，这取决于pH、试剂浓度、温度和孵育时间等实验条件。pH、适体浓度、竞争孵育时间、竞争孵育温度和酶标抗体浓度的优化有助于实现这些目标。实验结果表明，该传感器的检出限为0.067 ng/mL，在0.25 ~ 10 ng/mL的浓度范围内线性良好（R2 = 0.99679）。加样回收率为94.4% ~ 108.05%。该传感器具有检测限低、操作简单、成本低的特点，为开发具有成本效益的高精度霉菌毒素检测方法提供了一种新的解决方案。

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来源期刊

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.