食品科学 ›› 2011, Vol. 32 ›› Issue (11): 110-113.doi: 10.7506/spkx1002-6630-201111025

• 生物工程 • 上一篇    下一篇

抗大田软海绵酸单抗独特型抗体的制备及鉴定

林 超,王东旭,任洪林,柳增善,周 玉,李岩松,闫东明,王里奇,卢士英*   

  1. 人兽共患病研究教育部重点实验室,吉林大学人兽共患病研究所,吉林大学畜牧兽医学院
  • 出版日期:2011-06-15 发布日期:2011-05-13
  • 基金资助:
    国家自然科学基金项目(31071539;30671762);国家质检总局科技计划项目(2008IK003); 吉林大学农学部博士科研启动基金项目(4305050102C8)

Preparation and Identification of Anti-idiotypic Antibody against Anti-okadaic Acid Monoclonal Antibody

LIN Chao,WANG Dong-xu,REN Hong-lin,LIU Zeng-shan,ZHOU Yu,LI Yan-song, YAN Dong-ming,WANG Li-qi,LU Shi-ying*   

  1. Key Laboratory of Zoonosis Research, Ministry of Education, Institute of Zoonosis, College of Animal Science and Veterinary Medicine, Jilin University, Changchun 130062, China
  • Online:2011-06-15 Published:2011-05-13

摘要: 目的:研制大田软海绵酸(okadaic acid,OA)单抗的独特型抗体并鉴定其模拟抗原特性,为建立无毒检测提供借鉴。方法:利用小鼠腹水法大量制备抗OA单抗,经辛酸饱和硫酸铵法纯化后用胃蛋白酶酶切制备F(ab’)2 片段,免疫日本大耳白兔,取其血清,琼脂扩散法检测血清抗体与抗OA单抗反应活性, ELISA方法鉴定血清抗体与抗OA单抗反应活性及与OA竞争活性。结果:该抗独特型抗体与抗OA单抗呈阳性反应,且可抑制抗OA单抗与固相抗原OA-OVA的结合。结论:制备的抗OA单抗的独特型抗体与OA抗原存在内影像的关系,可替代毒素用于检测。

关键词: 大田软海绵酸(OA), F(ab’)2 片段, 抗独特型抗体, 内影像

Abstract: Objective: In order to develop a toxin-free detection method for okadaic acid (OA) from the shellfish, an anti-idiotypic antibody against anti-OA toxin monoclonal antibody (McAb) was prepared and identified. Methods: F(ab’)2 fragment from anti-OA McAb that was massively produced by the mouse ascites method and purified by caprylic acid and saturated ammonium sulfate was prepared by pepsin cleavage. Japanese big-ear white rabbits were immunized with the F(ab’)2 fragment and then their sera were taken for determination of reactivity between serum antibodies and anti-OA McA by agar gel diffusion test, indirect ELISA and indirect competitive ELISA. Results: The anti-idiotypic antibody had positive reaction with anti-OA McAb and competitively inhibited the binding of anti-OA McAb with detection antigen (OA-OVA). Conclusion: The prepared anti-idiotypic antibody against anti-OA monoclonal antibody has inner image of OA antigen and can be used to replace OA toxin standard in ELISA.

Key words: okadaic acid (OA), F(ab’)2 fragment, anti-idiotype antibody, inner image

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