Immunochemical authentication of manuka honey using a monoclonal antibody specific to a glycoside of methyl syringate
Immunochemical authentication of manuka honey using a monoclonal antibody specific to a glycoside of methyl syringate
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使用丁香酸甲酯糖苷特异性单克隆抗体对麦卢卡蜂蜜进行免疫化学认证
DOI：
10.1021/jf503464a
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发表时间：
2014
期刊：
影响因子：
--
通讯作者：
Yosuke Takimoto,
中科院分区：
文献类型：
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作者：
Yoji Kato;Yukako Araki;Maki Juri;Rie Fujinaka;Akari Ishisaka;Noritoshi Kitamoto;Yoko Nitta;Toshio Niwa;Yosuke Takimoto,
丁香酸甲酯糖苷是丁香酸甲酯的一种新型糖苷，仅存在于从丁香酸甲酯中提取的麦卢卡蜂蜜中。因此，钩端螺旋体的定量可能适用于蜂蜜的鉴定。钩端螺旋体的浓度与抗菌活性呈高度线性关系。我们建立了抗钩端螺旋体的单抗，并用竞争性酶联免疫吸附试验(ELISA)对该抗体进行了详细的鉴定，并与高效液相色谱(HPLC)方法的结果进行了比较以验证。麦卢卡蜂蜜中的抗原经高效液相色谱分离鉴定为钩端螺旋体蛋白，并用酶联免疫吸附试验进行定量。用建立的可在一个96孔板上处理20个样品(重复)的酶联免疫吸附试验对50个蜂蜜样品中的钩端螺旋体含量进行了分析。与化学定量结果有很好的一致性。对于麦卢卡蜂蜜的鉴定，钩端螺旋体免疫化学定量将是一种经济和简便的方法，允许多个蜂蜜样品同时处理和用ELISA进行分析。
Leptosperin, a novel glycoside of methyl syringate, is exclusively present in manuka honey derived from theLeptospermumspeciesLeptospermum scoparium. Quantification of leptosperin might thus be applicable for authentication of honey. The concentration of leptosperin has high linearity with antibacterial activity. We established a monoclonal antibody to leptosperin and characterized the antibody in detail by a competitive enzyme-linked immunosorbent assay (ELISA), comparing the results with those of the high-performance liquid chromatography (HPLC) method for validation. The antigen in manuka honey was confirmed as leptosperin by HPLC fractionation with quantitation by an ELISA. Leptosperin contents of 50 honey samples were analyzed by an established ELISA, which can handle 20 samples (duplicate) on one 96-well plate. Significant coincidence with the chemical quantitation was observed. Immunochemical quantitation of leptosperin would be an economical and facile method for the possible authentication of manuka honey, allowing many honey samples to be processed and analyzed by an ELISA simultaneously.