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B检测中的应用
Development of a Novel Aptamer-Antibody Sandwich Chemiluminescent Biosensor and Its Application in the Detection of Aflatoxin B.
作者信息
Zhao Zhike, Feng Jianghao, Wu Caizhang
机构信息
Key Laboratory of Grain Information Processing and Control, Henan University of Technology, Ministry of Education, Zhengzhou 450001, China.
Henan Key Laboratory of Grain Storage Information Intelligent Perception and Decision Making, Henan University of Technology, Zhengzhou 450001, China.
出版信息
Biosensors (Basel). 2025 Aug 15;15(8):538. doi: 10.3390/bios15080538.
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 B (AFB) was developed. The indirect competition between AFB, aflatoxin B-ovomucoid complete antigen (AFB-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 AFB. The biosensor was designed to operate with high precision, low cost, and a low detection limit for AFB, 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 (R = 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.
为应对霉菌毒素检测中高成本、低准确性和操作繁琐等挑战，开发了一种用于检测黄曲霉毒素B（AFB）的新型适体-抗体夹心化学发光生物传感器。AFB、黄曲霉毒素B-卵类粘蛋白完全抗原（AFB-OVA）和兔抗卵类粘蛋白（OVA）抗体之间的间接竞争导致形成夹心复合物。这种夹心测定法与辣根过氧化物酶标记的抗体相连，该抗体催化鲁米诺化学发光以间接检测AFB。该生物传感器设计用于高精度、低成本运行，且对AFB具有低检测限，这取决于诸如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、竞争孵育温度和辣根过氧化物酶标记抗体浓度的优化有助于实现这些目标。实验结果表明，该传感器的检测限为0.067 ng/mL，在0.25 - 10 ng/mL的浓度范围内表现出良好的线性（R = 0.99679）。加标样品的回收率在94.4%至108.05%之间。该传感器具有低检测限、操作简单和成本低的特点，从而为开发经济高效、高精度的霉菌毒素检测方法提供了一种新的解决方案。
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