中国畜牧兽医

• 生物技术 • 上一篇    下一篇

展青霉素人工抗原的制备与鉴定

王欣然1,聂虹霞2,李岩松1,谭彩云2,胡盼1,冯小丽1,任洪林1,郭一平2,周玉1,柳增善1,卢士英1   

  1. 1.吉林大学人兽共患病研究所,人兽共患病教育部重点实验室,吉林长春  1300622.大连众信检测技术有限公司,辽宁大连  116045

  • 修回日期:2014-04-02 出版日期:2014-07-20 发布日期:2014-08-21
  • 通讯作者: 卢士英(1973—),女,博士,副教授,研究方向:食品安全与人兽共患细菌病。Tel:0431-87836716;E-mail:lushiying1129@163.com
  • 作者简介:王欣然(1989—),女,吉林人,硕士生,研究方向:兽医公共卫生。
  • 基金资助:

    国家国际合作专项(2011DFA32230)。

Preparation and Identification of Artificial Antigen for Patulin

WANG Xin-ran1, NIE Hong-xia2, LI Yan-song1, TAN Cai-yun2, HU Pan1, FENG Xiao-li1, REN Hong-lin1, GUO Yi-ping2, ZHOU Yu1, LIU Zeng-shan1, LU Shi-ying1   

  1. 1. Key Laboratory for Zoonosis Research, Ministry of Education, Institute of Zoonosis, Jilin University, Changchun 130062, China; 2. Dalian Zhongxin Testing Technology, LLC, Dalian 116045, China)

  • Revised:2014-04-02 Online:2014-07-20 Published:2014-08-21

摘要: 为探讨展青霉素(patulin,PAT)不同抗原合成方法对完全抗原制备及抗体产生的影响,本试验利用琥珀酸酐联合活泼酯法,分别制备免疫原PAT-BSA和包被原PAT-OVA。为提高PAT与偶联蛋白的结合率,尝试将BSA修饰为EDA-BSA后与PAT进行偶联,并对比2种偶联方法制备的完全抗原免疫动物后抗血清的效价水平。同时利用琼脂糖凝胶电泳、非变性聚丙烯酰胺凝胶电泳、变性聚丙烯酰胺凝胶电泳、凝胶渗透色谱多种方法分析鉴定偶联结果,为PAT单克隆抗体的制备及免疫学检测方法的建立奠定了基础。

关键词:

展青霉素; 偶联; 鉴定; 完全抗原

Abstract: In order to explore the different antigen synthesis method influence on the preparation of complete antigen and the production of antibodies, the immunogen and coating antigen were synthesized by succinic anhydride and active ester method. Alternative protein carrier of EDA-BSA modifided from BSA was used for coupling the hapten to improve binding rate. Then comparing the difference of mice serum antibody titer immuned by two artificial antigens. Features of the conjugations were identified by agarose gels, native-PAGE, SDS-PAGE and GPC. The study would establish favourable foundation for preparation of antibody against PAT and immunological detecting method.

Key words:

patulin; coupling conjugate; identification; complete antigen