| 1-芘丁酸人工抗原的制备及鉴定 |
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| 引用本文： | 孔蒙蒙,黄忠民,邓瑞广,索标,刘情情,胡骁飞,王娜.1-芘丁酸人工抗原的制备及鉴定[J].食品与生物技术学报,2021,40(12):99-104. |
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| 作者姓名： | 孔蒙蒙 黄忠民 邓瑞广 索标 刘情情 胡骁飞 王娜 |
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| 作者单位： | 河南农业大学 食品科学技术学院，河南 郑州 450002; 河南省农业科学院 动物免疫学重点实验室，河南 郑州 450002; 农业农村部大宗粮食加工重点实验室，河南 郑州 450002;河南农业大学 食品科学技术学院，河南 郑州 450002; 农业农村部大宗粮食加工重点实验室，河南 郑州 450002;河南农业大学 食品科学技术学院，河南 郑州 450002; 河南省农业科学院 动物免疫学重点实验室，河南 郑州 450002; 郑州市营养与健康食品重点实验室，河南 郑州 450002;河南农业大学 食品科学技术学院，河南 郑州 450002; 河南省农业科学院 动物免疫学重点实验室，河南 郑州 450002; 农业农村部大宗粮食加工重点实验室，河南 郑州 450002; 郑州市营养与健康食品重点实验室，河南 郑州 450002 |
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| 摘 要： | 为制备及鉴定1-芘丁酸（1-pyrenebutyric acid）人工抗原。采用碳化二亚胺（EDC）法将1-芘丁酸与牛血清白蛋白（BSA）和鸡卵清白蛋白（OVA）进行偶联，合成人工免疫原PBA-BSA和人工检测抗原PBA-OVA；紫外扫描及SDS-聚丙烯凝胶电泳（SDS-PAGE）结果显示抗原制备成功；鼠源多抗血清的效价均已超过1 000，IC50=14.31 ng/mL；与多环芳烃1-芘甲醇、1-芘甲醛、芘的交叉反应率小于14.40%，与菲、萘、苯并芘、BSA以及OVA交叉反应率均小于0.05%。作者成功制备出了PBA-BSA抗原，并且得到了敏感性良好的鼠源多抗血清，为后期单克隆抗体的制备及免疫学快速检测方法的建立奠定基础。
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| 关 键 词： | 1-芘丁酸 鼠源多抗血清 人工抗原 交叉反应率 |
Preparation and Identification of 1-Pyrenebutyric Acid Artificial Antigen |
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| KONG Mengmeng,HUANG Zhongmin,DENG Ruiguang,SUO Biao,LIU Qingqing,HU Xiaofei,WANG Na.Preparation and Identification of 1-Pyrenebutyric Acid Artificial Antigen[J].Journal of Food Science and Biotechnology,2021,40(12):99-104. |
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| Authors: | KONG Mengmeng HUANG Zhongmin DENG Ruiguang SUO Biao LIU Qingqing HU Xiaofei WANG Na |
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| Affiliation: | College of Food Science and Technology, Henan Agricultural University, Zhengzhou 450002, China; Key Laboratory of Animal Immunology, Henan Academy of Agricultural Sciences, Zhengzhou 450002, China; Key Laboratory of Staple Grain Processing, Ministry of Agriculture and Rural Affairs, Zhengzhou 450002, China;College of Food Science and Technology, Henan Agricultural University, Zhengzhou 450002, China; Key Laboratory of Staple Grain Processing, Ministry of Agriculture and Rural Affairs, Zhengzhou 450002, China;College of Food Science and Technology, Henan Agricultural University, Zhengzhou 450002, China; Key Laboratory of Animal Immunology, Henan Academy of Agricultural Sciences, Zhengzhou 450002, China; Zhengzhou Nutrition and Health Food Laboratory, Zhengzhou 450002, China; College of Food Science and Technology, Henan Agricultural University, Zhengzhou 450002, China; Key Laboratory of Animal Immunology, Henan Academy of Agricultural Sciences, Zhengzhou 450002, China; Key Laboratory of Staple Grain Processing, Ministry of Agriculture and Rural Affairs, Zhengzhou 450002, China; Zhengzhou Nutrition and Health Food Laboratory, Zhengzhou 450002, China |
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| Abstract: | The preparation and identification of 1-pyrenebutyric acid artificial antigen was investigated in this paper.1-pyrenebutyric acid was coupled to bovine serum albumin (BSA) and chicken oval albumin (OVA) using the carbodiimide method (EDC) to synthesize the artificial immunogen PBA-BSA and artificial detection antigen PBA-OVA.The results of UV scanning and polypropylene gel electrophoresis (SDS-PAGE) showed that the antigen was successfully prepared. The potency of mouse-derived polyantisera was more than 1 000 with IC50 of 14.31 ng/mL. The cross-reactivity with polycyclic aromatic hydrocarbons 1-pyrene methanol, 1-pyrene formaldehyde and pyrene was less than 14.40%, and the cross-reactivity with phenanthrene, naphthalene, benzopyrene, BSA and OVA was less than 0.05%. PBA-BSA antigen was successfully prepared, and a sensitive murine multi antiserum was obtained, laying the foundation for the subsequent preparation of monoclonal antibodies and the establishment of rapid immunological detection methods. |
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| Keywords: | 1-pyrenebutyric acid mouse polyclonal antiserum artificial antigen cross reaction rate |
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