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基于新型多克隆抗体的快速金免疫渗滤层析法检测蜂蜜中主要蜂王浆蛋白 1(MRJP1)

Novel polyclonal antibody-based rapid gold sandwich immunochromatographic strip for detecting the major royal jelly protein 1 (MRJP1) in honey.

机构信息

Department of Food Science and Nutrition, Zhejiang Key Laboratory for Agro-Food Processing, Zhejiang University, Hangzhou, Zhejiang, China.

Qinhuangdao Entry-Exit Inspection and Quarantine Bureau, Qinhuangdao, Hebei, China.

出版信息

PLoS One. 2019 Feb 19;14(2):e0212335. doi: 10.1371/journal.pone.0212335. eCollection 2019.

Abstract

Honey adulteration is becoming increasingly alarming incidents in food safety. Monitoring and detecting adulteration face greater challenges. Honey contains the major royal jelly proteins (MRJP) secreted by bee workers. To detect honey adulteration fast and accurately, a rapid gold sandwich immunochromatographic strip (GSIS) was developed based on two specific polyclonal antibodies (PoAbs) against the MRJP1, the most abundant protein of all MRJPs. We determined the best of pH value (pH 8.6) and PoAb SP-1 amount (5 μg/mL) in conjunction with colloidal. The cut-off value (sensitivity) of GSIS in detecting MRJP1 is 2.0 μg/mL in solution. Validation analysis with RJ, milk vetch honey, acacia honey and honey adulteration containing rice syrup and corn syrup with different ratios demonstrated that the GSIS could show consistent Test line (T line) when the test samples contain more than 30% pure honey or MRJP1 0.4 mg/g. The validation results by isotope ratio mass spectrometry on the same pure and all adulteration milk vetch honey samples showed the same information of GSIS test. The qualitative assay GSIS provided a valuable new way for honey authenticity and laid the foundation for the future application of GSIS with monoclonal antibodies in honey authentication.

摘要

蜂蜜掺假是食品安全中日益令人担忧的事件。监测和检测掺假面临更大的挑战。蜂蜜中含有蜜蜂工蜂分泌的主要蜂王浆蛋白(MRJP)。为了快速准确地检测蜂蜜掺假,我们基于针对 MRJP1 的两种特异性多克隆抗体(PoAb)开发了一种快速金夹心免疫层析条(GSIS),MRJP1 是所有 MRJPs 中含量最丰富的蛋白质。我们确定了最佳 pH 值(pH 8.6)和胶体结合时的 PoAb SP-1 量(5 μg/mL)。在溶液中,GSIS 检测 MRJP1 的截断值(灵敏度)为 2.0 μg/mL。用 RJ、黄芪蜜、刺槐蜜和含有不同比例大米糖浆和玉米糖浆的蜂蜜掺假进行验证分析表明,当测试样品中含有超过 30%的纯蜂蜜或 0.4 mg/g 的 MRJP1 时,GSIS 可以显示一致的测试线(T 线)。对相同的纯品和所有掺假黄芪蜜样品进行同位素比质谱验证的结果表明,GSIS 测试的信息相同。定性分析 GSIS 为蜂蜜真实性提供了一种有价值的新方法,并为未来在蜂蜜鉴定中使用单克隆抗体进行 GSIS 应用奠定了基础。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1cb8/6380560/7f3f68deb14a/pone.0212335.g001.jpg

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