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用于检测古代羊毛的酶联免疫吸附测定法和金标免疫层析试纸条测定法的开发

Development of an Enzyme-Linked Immunosorbent Assay and Gold-Labelled Immunochromatographic Strip Assay for the Detection of Ancient Wool.

作者信息

Wang Bing, Gu Jincui, Chen Boyi, Xu Chengfeng, Zheng Hailing, Peng Zhiqin, Zhou Yang, Hu Zhiwen

机构信息

Key Laboratory of Advanced Textile Materials and Manufacturing Technology, Ministry of Education, Zhejiang Sci-Tech University, Hangzhou 310018, China.

Key Scientific Research Base of Textile Conservation, State Administration for Cultural Heritage, China National Silk Museum, Hangzhou 310002, China.

出版信息

J Anal Methods Chem. 2018 Jun 5;2018:2641624. doi: 10.1155/2018/2641624. eCollection 2018.

Abstract

The identification of ancient wool is of great importance in archaeology. Despite lots of meaningful information can be achieved by conventional detection methods, that is, light and electron microscopy, spectroscopy, and chromatography, the efficacy is likely to be limited in the detection of ancient samples with contamination or severe degradation. In this work, an immunoassay was proposed and performed for the identification of ancient wool. First, a specific antibody, which has the benefits of low cost, easy operation, and extensive applicability, was developed directly through immunizing rabbits with complete antigen (keratin). Then, an enzyme-linked immunosorbent assay (ELISA) and a colloidal gold-labelled immunochromatographic strip (ICS) were developed to qualitatively identify the corresponding protein in ancient wool samples unearthed from Kazakhstan and China. The anti-keratin antibody exhibited high sensitivity and specificity for the identification of modern and ancient wool. The limit of detection (LOD) of the ELISA method was 10 ng/mL, and no cross-reactions with other interfering antigens have been noted. It is concluded that the immunoassays are reliable methods for the identification of ancient wool.

摘要

古代羊毛的鉴定在考古学中具有重要意义。尽管通过传统检测方法,即光学显微镜和电子显微镜、光谱学和色谱法,可以获得许多有意义的信息,但在检测受污染或严重降解的古代样本时,其效果可能会受到限制。在这项工作中,提出并进行了一种免疫测定法用于古代羊毛的鉴定。首先,通过用完全抗原(角蛋白)免疫兔子,直接制备了一种具有低成本、操作简便和广泛适用性等优点的特异性抗体。然后,开发了酶联免疫吸附测定法(ELISA)和胶体金标记免疫层析试纸条(ICS),用于定性鉴定从哈萨克斯坦和中国出土的古代羊毛样本中的相应蛋白质。抗角蛋白抗体在鉴定现代和古代羊毛时表现出高灵敏度和特异性。ELISA方法的检测限为10 ng/mL,且未观察到与其他干扰抗原的交叉反应。得出结论,免疫测定法是鉴定古代羊毛的可靠方法。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/eae4/6008753/bbd0afed52ce/JAMC2018-2641624.001.jpg

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