中国畜牧兽医

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猪IL-15多克隆抗体的制备及其生物活性的检测

邹雨妃, 王衡, 远立国, 李守军   

  1. 华南农业大学兽医学院,广东广州 510642
  • 收稿日期:2012-02-14 出版日期:2012-09-20 发布日期:2012-09-18
  • 通讯作者: 王衡(1981-),男,黑龙江人,讲师,博士,从事兽医病理学研究。E-mail:wangheng2009@scau.edu.cn E-mail:wangheng2009@scau.edu.cn
  • 作者简介:邹雨妃(1989-),女,江西人,硕士生,研究方向:临床兽医学。
  • 基金资助:

    "大连三仪-华南农大科研基金"资助项目;高等学校博士点学科专项科研基金(20114404120010)。

Preparation and Biological Activity Analysis of the Polyclone Antibody to Porcine IL-15

ZOU Yu-fei, WANG Heng, YUAN Li-guo, LI Shou-jun   

  1. College of Veterinary Medicine,South China Agricultural University,Guangzhou 510642,China
  • Received:2012-02-14 Online:2012-09-20 Published:2012-09-18

摘要: 猪IL-15与人IL-15具有较高的相似率,为进一步研究其生物活性,本研究将去除信号肽的猪IL-15克隆至原核表达载体pET-30a,并成功获得其重组蛋白。将纯化的重组蛋白用于多克隆抗体的制备,并利用琼脂扩散方法、ELISA方法、Western blotting试验及免疫荧光试验对其抗体效价、特异性、灵敏度等生物活性进行了检测。结果表明,该抗体具有较高的抗体效价和很好的特异性,可以作为IL-15特异性抗体用于今后的试验研究。

关键词: 猪IL-15; 多克隆抗体; 生物活性

Abstract: Porcine IL-15 has a high similarity to human IL-15. Porcine IL-15 without signal peptide was cloned into prokaryotic expression vector pET-30a,the recombinant plasmid was transformed into E.coli BL21,and induced by IPTG. Recombinant protein was aquired successfully,then the purified protein was used to prepare polyclone antibody. Agarose diffusion assay,ELISA,Western blotting assy and immunofluorescence were used to detect the titer,specificity and sensitvity of the antibody. The results revealed that the polyclone antibody prepared in this study had a high titer and good specificity,and it could be used as the specific antibody to porcine IL-15 for the future research.

Key words: porcine IL-15; polyclonal antibody; biological activity

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