中国畜牧兽医

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大观霉素人工抗原的合成及其多克隆抗体制备

王照鹏1,李向梅1,2,徐飞2,王文珺1,苏丽芳1,温凯2,吴小平1   

  1. 1.北京维德维康生物技术有限公司,北京 100095;2.中国农业大学动物医学院,北京 100193)
  • 收稿日期:2013-07-04 出版日期:2014-02-20 发布日期:2014-03-27
  • 作者简介:王照鹏(1983—),男,山东人,硕士,研究方向:动物性食品中兽药安全残留检测。
  • 基金资助:

    国家十二五科技支撑计划项目(2011BAK10B01-04)。

Synthesis of Artificial Antigen of Spectinomycin and Preparation of its Polyclonal Antibody

WANG Zhao-peng1, LI Xiang-mei1,2, XU Fei2, WANG Wen-jun1, SU Li-fang1, WEN Kai2, WU Xiao-ping1   

  1. 1.Beijing WDWK Biotechnology Co., Ltd., Beijing 100095, China; 2.College of Veterinary Medicine, China Agricultural University, Beijing 100193, China)
  • Received:2013-07-04 Online:2014-02-20 Published:2014-03-27

摘要: 以大观霉素(SPE)、盐酸羧甲基羟胺(CMO)、乙二醇二环氧甘油醚(EDE)、牛血清白蛋白(BSA)、鸡卵清白蛋白(OVA)等为原料,采用活泼酯法和EDE双功能团活泼试剂方法分别合成2种免疫原和2种包被原。以免疫原免疫家兔制备多克隆抗体,以建立的间接ELISA方法来测定多克隆抗体效价及灵敏度。结果表明,免疫原SPE-CMO-BSA具有良好的免疫原性,制备的多克隆抗体经ELISA检测效价可达1∶160000,IC50为0.484 ng/mL。本研究结果为下一步SPE残留快速检测试剂盒的研制奠定了基础。

关键词: 大观霉素; 免疫原; 多克隆抗体; 效价

Abstract: Using spectinomycin (SPE), carboxyl methyl hydroxylamine hydrochloride (CMO), glycol diglycidyl ether (EDE), bovine serum albumin (BSA) and ovalbumin (OVA) as the main stuff, four artificial-immunogens had been obtained by the N-hydroxysuccinimide-activated ester method and the EDE double functional group method. Ten female rabbits were immunized with this two artificial-immunogens. Animal immune test demonstrated that there were antibodies against SPE in rabbits sera. The polyclonal antibody was measured with indirect ELISA test. The titer was 1∶160000 and the IC50 was 0.484 ng/mL. This study laid the basis for further research on the immune test kit for the SPE remnant.

Key words: spectinomycin; immunogen; polyclonal antibody; titer