松香人工抗原的合成及多克隆抗体的制备  

Synthesis of Artificial Antigens and Preparation of Multi-Clonal Antibody Against Rosin

作　　者：卞欢[1] 仇新媛 李鹏鹏[1] 陈琳[1] 耿志明[1] 王道营[1] 徐为民[1,3] BIAN Huan;QIU Xinyuan;LI Pengpeng;CHEN Lin;GENG Zhiming;WANG Daoying;XU Weimin(Institute of Agri-Products Processing,Jiangsu Academy of Agricultural Sciences,Nanjing 210014,China;College of Biotechnology and Pharmaceutical Engineering,Nanjing Tech University,Nanjing 211816,China;Jiangsu Collaborative Innovation Center of Meat Production and Processing,Quality and Safety Control,Nanjing 210095,China)

机构地区：[1]江苏省农业科学院农产品加工研究所,江苏南京210014 [2]南京工业大学生物与制药工程学院,江苏南京211816 [3]江苏省肉类生产与加工质量安全控制协同创新中心,江苏南京210095

出　　处：《食品科学技术学报》2018年第6期51-57,共7页Journal of Food Science and Technology

基　　金：江苏省农业科技自主创新资金项目(CX(16)1061)

摘　　要：利用脱氢松香胺(DHAM)为半抗原合成了松香的完全抗原、并通过免疫新西兰大白兔制备了多克隆抗体。首先DHAM与琥珀酸酐(SUC)反应合成琥珀酸单酰脱氢松香胺(DHAM-SUC),然后再与牛血清蛋白(BSA)和钥孔血蓝蛋白(KLH)进行偶联,合成了松香的完全抗原DHAM-SUCBSA和DHAM-SUC-KLH,偶联比分别为12和35。以DHAM-SUC-KLH为免疫原对新西兰大白兔进行免疫,制备了抗血清,效价达1.28×10~5,脱氢松香酸(DHA)和松香酸(AA)的50%抑制浓度(IC_(50))分别为25. 1μg/L和36. 4μg/L。研究合成的松香人工抗原具有理想的免疫原性,获得的多克隆抗体具有较好的灵敏度,可为建立畜禽肉制品中松香残留的酶联免疫吸附分析(ELISA)方法打下基础。Artificial antigens against rosin were synthesised with dehydroabietylamine（DHAM） as a hapten, and the multi-clonal antibody was prepared by immunizing New Zealand rabbits. At first, DHAM was reacted with succinic anhydride （SUC） followed by being conjugated to bull serum albumin （BSA） and keyhole limpet haemocyanin（KLH）. The artificial antigens （DHAM-SUC-BSA and DHAM-SUC-KLH） were synthesised, and the ratios of conjugation were 21 and 35. Multi-clonal antibody was raised in New Zealand rabbits by injection of DHAM-SUC-KLH, and the titre reached 1.28×10^5 . The IC 50 of dehydroabietic acid （DHA） and abietic acid （AA） were 25.1 μg/L and 36.4 μg/L. The result indicated that the immune antigen was a desired immunogenicity and the multi-clonal antibody showed a good sensitivity against DHA, which paved a way for building enzyme-linked immunosorbent assay （ELISA） of rosin residue in animal products.

关 键 词：松香 脱氢松香胺 人工抗原 多克隆抗体 酶联免疫吸附分析方法 

分 类 号：TS207.7[轻工技术与工程—食品科学]