›› 2011, Vol. 38 ›› Issue (6): 91-94.

• 生物技术 • 上一篇    下一篇

兔抗鹅IgG酶标抗体的制备及初步应用

刘菲1,2, 唐卫杰1, 韩清林1,3, 赵婷婷1, 高丽芹1, 严晓玲1   

  1. 1. 四川农业大学动物医学院,四川雅安 625014;2. 动物疫病与人类健康四川省重点实验室,四川雅安 625014;3. 四川省大英县畜牧局,四川大英 629300
  • 收稿日期:1900-01-01 修回日期:2011-03-27 出版日期:2011-06-20 发布日期:2011-06-20

Preparation of Rabbit Anti-goose IgG-HRP and Preliminary Application

LIU Fei1,2, TANG Wei-jie1, HAN Qing-lin1,3, ZHAO Ting-ting1, GAO Li-qin1, YAN Xiao-ling1   

  1. 1. College of Veterinary Medicine, Sichuan Agricultural University, Ya’an 625014, China;2. Key Laboratory of Animal Disease and Human Health of Sichuan Province, Ya’an 625014,China;3. Bureau of Animal Husbandry of Daying, Sichuan Province, Daying 629300, China
  • Received:1900-01-01 Revised:2011-03-27 Online:2011-06-20 Published:2011-06-20

摘要: 为了研究兔抗鹅IgG酶标抗体的制备,试验采用水稀释-辛酸-硫酸铵法粗提,透析除盐,DEAEA-50纤维素层析相结合的方法提取鹅卵黄IgG,经核酸蛋白检测仪和SDS-PAGE测定IgG浓度;制备兔抗鹅IgG酶标抗体;利用酶标抗体检测小鹅瘟病毒(GPV)VP3基因疫苗的免疫效果。结果表明,粗提的IgG浓度为10 mg/mL,抗体纯度可达到94.5%;免疫扩散得出兔抗鹅 IgG抗血清效价约 1∶32,抗体酶标后效价为1∶3000;酶标抗体可以作为检测疫苗效果一种很好的工具。

关键词: 鹅卵黄IgG; 提取; 酶标; 小鹅瘟VP3基因疫苗

Abstract: To study on preparation of rabbit anti-goose IgG-HRP, the experimentation isolated IgG from the goose egg yolk, and the method was employed to dilute with water-n-octylic acid-ammonia sulfate initial extraction, out salt dialysis and DEAEA-50 chromatography. Protein nucleic acid detector and SDS-PAGE indicated the IgG concentration; the purified IgG was labeled with HRP; effect of GPV VP3 gene vaccine was detected with the labeled antibody. The results indicated that the initial extraction was 10 mg/mL and the purity was 94.5%; the antiserum characterized by immunodiffusion showed that the titer of the antiserum was 1∶32 and specificity of the antiserum was high. The titer of labeled antibody was 1∶3000; the labeled antibody was a good tool to detect the effect of GPV VP3 gene vaccine.

Key words: goose egg yolk IgG; isolation; enzyme labeling; pcDNA-VP3 DNA vaccine

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