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基于荧光破伤风人IgG-金纳米簇的免疫层析试纸条检测破伤风类毒素

Detection of tetanus toxoid with fluorescent tetanus human IgG-AuNC-based immunochromatography test strip.

作者信息

Zhuang Quan-Quan, Chen Rui-Ting, Zheng Yi-Jing, Huang Kai-Yuan, Peng Hua-Ping, Lin Zhen, Xia Xing-Hua, Chen Wei, Deng Hao-Hua

机构信息

Higher Educational Key Laboratory for Nano Biomedical Technology of Fujian Province, Department of Pharmaceutical Analysis, Fujian Medical University, Fuzhou, 350004, China; Department of Pharmacy, Affiliated Quanzhou First Hospital of Fujian Medical University, Quanzhou, 362000, China.

Higher Educational Key Laboratory for Nano Biomedical Technology of Fujian Province, Department of Pharmaceutical Analysis, Fujian Medical University, Fuzhou, 350004, China.

出版信息

Biosens Bioelectron. 2021 Apr 1;177:112977. doi: 10.1016/j.bios.2021.112977. Epub 2021 Jan 6.

Abstract

Assays for detecting tetanus toxoid are of great significance to be applied in the research of the safety testing of tetanus vaccine. Currently, guinea pigs or mice are usually used to evaluate the toxicity in these assays. Herein, a facile and quick biomineralization process was carried out to generate tetanus human immunoglobulin G (Tet-IgG)-functionalized Au nanoclusters (Tet-IgG-AuNCs). The obtained Tet-IgG-AuNCs exhibited strong red emission with a photoluminescence quantum yield of 13%. Based on surface plasmon resonance measurements, the apparent dissociation constant of the Tet-IgG-AuNC-tetanus toxoid complexes was measured to be 2.27 × 10 M. A facile detection approach was developed using a fluorescent Tet-IgG-AuNC-based immunochromatography test strip. By utilizing the high-brightness fluorescent Tet-IgG-AuNCs, this immunosensor showed favorable sensitivity with a detection limit at the level of 0.03 μg/mL. Further results demonstrated that this assay can reliably detect tetanus toxoid and therefore might provide a novel method to replace animal tests for the quantification of tetanus toxicity. Moreover, the antibody-AuNC-based immunochromatography test strip platform serves as a promising candidate to develop new approaches for detecting targeted antigens and biological events of interest.

摘要

检测破伤风类毒素的分析方法在破伤风疫苗安全性测试研究中具有重要意义。目前,在这些分析中通常使用豚鼠或小鼠来评估毒性。在此,开展了一个简便快速的生物矿化过程,以生成破伤风人免疫球蛋白G(Tet-IgG)功能化的金纳米簇(Tet-IgG-AuNCs)。所获得的Tet-IgG-AuNCs呈现出强烈的红色发射,光致发光量子产率为13%。基于表面等离子体共振测量,测得Tet-IgG-AuNC-破伤风类毒素复合物的表观解离常数为2.27×10⁻⁷ M。利用基于荧光Tet-IgG-AuNC的免疫层析试纸条开发了一种简便的检测方法。通过利用高亮度荧光Tet-IgG-AuNCs,这种免疫传感器显示出良好的灵敏度,检测限为0.03 μg/mL。进一步的结果表明,该分析方法能够可靠地检测破伤风类毒素,因此可能提供一种替代动物试验来定量破伤风毒性的新方法。此外,基于抗体-金纳米簇的免疫层析试纸条平台有望成为开发检测目标抗原和感兴趣生物事件新方法的候选者。

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