食品科学 ›› 2013, Vol. 34 ›› Issue (7): 227-230.doi: 10.7506/spkx1002-6630-201307047

• 生物工程 • 上一篇    下一篇

隐性孔雀石绿多克隆抗体的制备及鉴定

梁莉甜,刘志国*,付云洁,尹 欢,马寅众,李 奎   

  1. 武汉工业学院生物与制药工程学院,湖北 武汉 430023
  • 收稿日期:2012-04-15 修回日期:2013-02-23 出版日期:2013-04-15 发布日期:2013-03-20
  • 通讯作者: 刘志国 E-mail:zhiguo_l@126.com
  • 基金资助:
    武汉市科技局对外科技合作与交流计划项目(201070934341);武汉市科技局现代农业技术创新平台项目(201120637175-5); 武汉工业学院研究生创新基金项目(2011cx018)

Preparation and Characterization of Anti-leucomalachite Green Polyclonal Antibody

LIANG Li-tian,LIU Zhi-guo*,FU Yun-jie,YIN Huan,MA Yin-zhong,LI Kui   

  1. School of Biology and Pharmaceutical Engineering, Wuhan Polytechnic University, Wuhan 430023, China
  • Received:2012-04-15 Revised:2013-02-23 Online:2013-04-15 Published:2013-03-20
  • Contact: Zhi-Guo LIU E-mail:zhiguo_l@126.com

摘要: 研究制备孔雀石绿(MG)代谢物隐性孔雀石绿(LMG)多克隆抗体进而间接鉴定鱼体中孔雀石绿的残留。首先利用化学方法合成含羧基的隐性孔雀石绿衍生物(CLMG),然后采用碳化二亚胺法偶联牛血清白蛋白(BSA)合成免疫抗原隐性孔雀石绿-BSA,按照常规免疫程序免疫新西兰大白兔制备抗隐性孔雀石绿多克隆抗体。采用间接酶联免疫吸附实验(iELISA)测定抗血清中抗隐性孔雀石绿多克隆抗体效价。结果表明:抗隐性孔雀石绿的抗体效价为1: 32000,可用于隐性孔雀石绿的免疫学检测。

关键词: 隐性孔雀石绿, 免疫抗原, 多克隆抗体, 酶联免疫吸附

Abstract: Preparation of anti-leucomalachite green polyclonal antibody allows indirect determination of leucomalachite green (LMG), a major metabolite of malachite green (MG). Carboxyleucomalachite green (CLMG) was chemically synthesized and then coupled to bovine serum albumin (BSA) by the carbodiimide method so as to obtain leucomalachite green-BSA immunogen. The immunoantigen was injected into rabbits according to routine immunity procedure to product anti-leucomalachite green antibody. The specific antibody of leucomalachite green was identified by indirect enzyme-linked immunosorbent assay (iELISA). It was proved that the working concentration of the specific antibody was 1: 32000 and therefore could be applied for the immunodetection of leucomalachite green.

Key words: leucomalachite green, immunoantigen, polyclonal antibody, ELISA

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