[1]鲁 絮,张彦明*,郑增忍,等.抗猪瘟病毒NS3(p80)蛋白单克隆抗体的制备及其特性鉴定[J].中国预防兽医学报,2008,(06):464-468.
 LU Xu,ZHANG Yan-ming*,ZHENG Zeng-ren,et al.Preparation of monoclonal antibodies against NS3 (p80) protein of CSFV[J].Chinese journal of preventive veterinary medicine,2008,(06):464-468.
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抗猪瘟病毒NS3(p80)蛋白单克隆抗体的制备及其特性鉴定
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《中国预防兽医学报》[ISSN:1008-0589/CN:23-1417/S]

卷:
期数:
2008年06期
页码:
464-468
栏目:
免疫学
出版日期:
2008-06-01

文章信息/Info

Title:
Preparation of monoclonal antibodies against NS3 (p80) protein of CSFV
文章编号:
1008-0589(2008)06-0464-05
作者:
鲁 絮12张彦明1*郑增忍2穆 杨1龚振华2李 葳2徐 浩1余 欣1
1. 西北农林科技大学 动物医学院,陕西 杨凌 712100;
2. 中国动物卫生与流行病学中心,山东 青岛 266032
Author(s):
LU Xu12 ZHANG Yan-ming1* ZHENG Zeng-ren2 MU Yang1 GONG Zhen-hua2 LI Wei2 XU Hao1 YU Xin1
1. College of Veterinary Medicine, Northwest Agricalture and Forestry University, Yangling 712100, China;
2. China Animal Health and Epidemiology Center, Qingdao 266032, China
关键词:
猪瘟病毒NS3基因表达蛋白纯化单克隆抗体
Keywords:
classical swine fever virus (CSFV) NS3 gene expression protein purification monoclonal antibody (McAb)
分类号:
S852.651
文献标志码:
A
摘要:
在大肠杆菌Rosetta (DE3)中表达猪瘟病毒石门株NS3基因部分功能片段,以纯化的重组NS3 (p80)蛋白免疫BALB/c小鼠,取其脾细胞与骨髓瘤细胞SP2/0进行融合,经间接ELISA筛选及3次连续克隆化获得了4株可分泌抗NS3单抗的杂交瘤细胞(2G7,2F11,5D2和6F11),通过小鼠体内诱生法制备腹水,并对杂交瘤细胞和单克隆抗体的特性进行鉴定。结果表明,杂交瘤细胞染色体正常,抗体分泌稳定,单抗效价高,亲和力各不相同,Ig亚类鉴定均为IgG1。Western blot分析证明4株单抗均能与原核表达的NS3蛋白特异性结合,6F11株与真核表达的NS3蛋白有较强的特异性反应。
Abstract:
Monoclonal antibodies against NS3 gene of CSFV was developed using recombinant NS3 (p80) protein expressed in E.coli Rosetta (DE3) as antigen. BALB/c mice was immunized with purified NS3 (p80) protein and mouse splenic cells were fused with SP2/0 cells. Hybridoma cells were screened by ELISA, subcloned and characterized. Four hybridoma cells (2G7, 2F11, 5D2 and 6F11) secreting anti-NS3 monoclonal antibodies (McAbs) were obtained. Hybridoma cells with normal chromosome were shown to stably secret NS3 specific antibodies with various affinities. All the McAbs belong to IgGl subtype.

参考文献/References:

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备注/Memo

备注/Memo:
收稿日期:2007-09-13
基金项目:国家自然科学基金项目(30270988);陕西省自然科学基金项目(2006C121)
作者简介:鲁 絮(1983~),女,陕西西安人,硕士,主要从事分子病原学与免疫学研究.
*通讯作者:E-mail:ylzhangym@sohu.comzhangym@nwsuaf.edu.cn
更新日期/Last Update: 2010-12-23