›› 2014, Vol. 41 ›› Issue (11): 97-100.

• 生物技术 • 上一篇    下一篇

抗猪瘟病毒单克隆抗体的制备及特性鉴定

王爱华1, 许可军2, 王金良3, 魏凤3, 孙全文1   

  1. 1. 河北北方学院, 河北张家口 075000;
    2. 滨州市农村科技信息化管理中心, 山东滨州 256600;
    3. 山东省滨州畜牧兽医研究院, 山东滨州 256600
  • 收稿日期:2014-06-03 出版日期:2014-11-20 发布日期:2014-12-06
  • 通讯作者: 王金良 E-mail:wjl478@163.com
  • 作者简介:王爱华(1966-),女,河北人,博士,教授,研究方向:动物疫病检验与公共卫生。
  • 基金资助:
    河北北方学院重大项目(ZD201307);山东省自然科学基金项目(ZR2010CQ012);山东省大型科学仪器设备升级改造技术研究专项(2013SJGZ18)。

Preparation and Characteristic Identification of Monoclonal Antibodies against Classical Swine Fever Virus

WANG Ai-hua1, XU Ke-jun2, WANG Jin-liang3, WEI Feng3, SUN Quan-wen1   

  1. 1. Hebei North University, Zhangjiakou 075000, China;
    2. Binzhou Rural Science and Technology Information Management Center, Binhzou 256600, China;
    3. Shandong Binzhou Animal Science and Veterinary Medicine Academy, Binzhou 256600, China
  • Received:2014-06-03 Online:2014-11-20 Published:2014-12-06

摘要: 将纯化后的猪瘟病毒免疫4只SPF级BALB/c小鼠,无菌取脾细胞与SP2/0骨髓瘤细胞融合,经半固体培养基克隆化和间接ELISA筛选,最终获得了4株稳定分泌抗猪瘟病毒单克隆抗体的杂交瘤细胞株。ELISA结果显示4株单克隆抗体效价均较高,并与其他病毒及PK-15细胞培养物无交叉反应。Western blotting结果证实3株单克隆抗体可特异性识别猪瘟病毒。间接免疫荧光试验可在细胞膜内观察到特异性绿色荧光,表明3株单克隆抗体与猪瘟病毒具有良好的反应性。该研究为猪瘟病毒新型诊断试剂开发奠定了基础。

关键词: 猪瘟病毒; 单克隆抗体; 制备; 鉴定

Abstract: 4 SPF BALB/c mice were immunized with purified classical swine fever virus (CSFV) as the antigen and mouse splenic cells were fused with SP2/0 cells. Hybridoma cells were screened by ELISA and semi-solid agar cloning, 4 hybridoma cells secreting anti-CSFV monoclonal antibodies were obtained. ELISA results showed that all of these monoclonal antibodies with high antibody titer, which could specifically react to CSFV but not cross react to other pig viruses and PK-15 cell cultures. Western blotting indicated that 3 strains of monoclonal antibodies reacted with CSFV specially. Specific green fluorescence was observed in the cell membrane by IFA. This study layed a foundation for the development of new diagnostic reagents for CSFV.

Key words: classical swine fever virus (CSFV); monoclonal antibody; preparation; identification

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