[1]张志榜,陈建飞,时洪艳,等.猪流行性腹泻病毒M蛋白单克隆抗体的制备及鉴定[J].中国预防兽医学报,2011,(07):568-570.
 ZHANG Zhi-bang,CHEN Jian-fei,SHI Hong-yan,et al.Preparation of the monoclonal antibody against membrane protein of porcine epidemic diarrhea virus[J].Chinese journal of preventive veterinary medicine,2011,(07):568-570.
点击复制

猪流行性腹泻病毒M蛋白单克隆抗体的制备及鉴定
分享到:

《中国预防兽医学报》[ISSN:1008-0589/CN:23-1417/S]

卷:
期数:
2011年07期
页码:
568-570
栏目:
研究简报
出版日期:
2011-07-15

文章信息/Info

Title:
Preparation of the monoclonal antibody against membrane protein of porcine epidemic diarrhea virus
作者:
张志榜;陈建飞;时洪艳;杨斌;石达;陈小金;冯力;
中国农业科学院哈尔滨兽医研究所兽医生物技术国家重点实验室/猪传染病研究室;内蒙古农业大学兽医学院;
Author(s):
ZHANG Zhi-bang1 CHEN Jian-fei1 SHI Hong-yan1 YANG Bin2 SHI Da1 CHEN Xiao-jin1 FENG Li1
1. State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin 150001, China; 2. College of Veterinary Medicine, Inner Mongolia Agricultural University, Huhhot 010018, China
关键词:
猪流行性腹泻病毒M蛋白单克隆抗体
Keywords:
porcine epidemic diarrhea virus M protein monoclonal antibody
摘要:
为制备猪流行性腹泻病毒(PEDV)抗M蛋白单克隆抗体(MAb),本研究以截短表达的His-M重组蛋白免疫BALB/c小鼠;以截短表达的GST-M重组蛋白作为包被抗原,采用常规的淋巴细胞杂交瘤技术制备杂交瘤细胞,通过间接ELISA进行筛选,得到一株稳定分泌抗M蛋白MAb。MAb亚类鉴定为IgG2b型,轻链为κ链,杂交瘤细胞培养上清和诱导的小鼠腹水抗体效价分别为1∶3000和1∶2×105。Westernblot试验表明该MAb能够识别重组及天然的PEDVM蛋白。间接免疫荧光试验表明该MAb能够与PEDV感染的VeroE6细胞产生特异性免疫荧光。
Abstract:
To prepare the monoclonal antibody (MAb) against of porcine epidemic diarrhea virus (PEDV), hybridomas were produced by fusing SP2/0 cells with spleen cells from mouse immunized with PEDV His-M recombinant protein. A hybridoma stable secreting MAb against M protein was identified by indirect ELISA detection with GST-M as coating antigen. The MAb was IgG2b subtype with κ chain. The titers in cell culture medium of the hybridoma and the ascetic fluids were 1:3,000 and 1:200,000, respectively. Western blot analysis showed that the MAb could recognize the recombinant and authentic M protein of PEDV, and the specific immunoflurescence was detected in PEDV infected Vero E6 cells by in direct immunoflurescence assay

参考文献/References:

[1] de Haan C A,Kuo L,Master P S,et al. Coronavirus particle assembly: Primary structure requirements of the membrane protein. Journal of Virology. 1998
[2] H Laude,J Gelfi,L Lavenant,B Charley. Single amino acid changes in the viral glycoprotein M affect induction of alpha interferon by the coronavirus transmissible gastroenteritis virus. Journal of Virology. 1992
[3] Shenyang G,Enhui Z,Baoxian Let al. High-level prokaryotic expression of envelope exterior of membrane protein of porcine epidemic diarrhea virus[J]. Veterinary Microbiology. 2007
[4] Kim O,Chae C. In situ hybridization for the detection and localization of porcine epidemic diarrhea virus in the intestinal tissues from naturally infected piglets. . 2000
[5] 陈建飞,冯力,时洪艳,孙东波,白兴华,佟有恩. 猪流行性腹泻病毒CH/S株N蛋白基因的遗传变异及其原核表达[J]. 中国预防兽医学报. 2007年11期
[6] 孙东波,冯力,时洪艳,陈建飞,崔晓辰,佟有恩. 猪流行性腹泻病毒S蛋白中和表位区单克隆抗体的制备与鉴定[J]. 中国预防兽医学报. 2007年11期

相似文献/References:

[1]周艳君,田志军,刘金霞,等.猪繁殖与呼吸综合征病毒M蛋白B细胞抗原表位的鉴定[J].中国预防兽医学报,2009,(04):251.
 ZHOU Yan-jun,TIAN Zhi-jun,LIU Jin-xia,et al.Identification of a B cell epitope on matrix protein of porcine reproductive and respiratory syndrome virus[J].Chinese journal of preventive veterinary medicine,2009,(07):251.
[2]葛俊伟,姜艳平,汪 淼,等.猪流行性腹泻病毒S1蛋白在干酪乳杆菌中的分泌表达及免疫原性分析[J].中国预防兽医学报,2009,(04):256.
 GE Jun-wei,JIANG Yan-ping,WANG Miao,et al.Recombinant Lactobacillus casei secretory expression of porcine epidemic diarrhea virus spike protein and the immunogenicity analysis[J].Chinese journal of preventive veterinary medicine,2009,(07):256.
[3]张维军,田 野,林 燕,等.猪繁殖与呼吸综合征病毒M蛋白在重组牛痘病毒中的表达与鉴定[J].中国预防兽医学报,2010,(10):741.[doi:doi: 10.3969/j.issn.1008-0589.2010.10.01]
 ZHANG Wei-jun,TIAN Ye,LIN Yan,et al.Construction of recombinant vaccinia virus expressing porcine reproductive and respiratory syndrome virus M protein[J].Chinese journal of preventive veterinary medicine,2010,(07):741.[doi:doi: 10.3969/j.issn.1008-0589.2010.10.01]
[4]姜艳平,葛俊伟,汪 淼,等.猪流行性腹泻病毒重组核蛋白作为检测抗原的初步应用[J].中国预防兽医学报,2008,(08):587.
 JIANG Yan-ping,GE Jun-wei,WANG Miao,et al.Preliminary application of the recombinant nucleocapsid protein of porcine epidemic diarrhea virus LJB/03 as detecting antigen in diagnostic assays[J].Chinese journal of preventive veterinary medicine,2008,(07):587.
[5]冯 力,孙东波,陈建飞,等.猪流行性腹泻病毒N蛋白单克隆抗体的制备与鉴定[J].中国预防兽医学报,2008,(03):190.
 FENG Li,SUN Dong-bo,CHEN Jian-fei,et al.Preparation and characterization of the monoclonal antibodies against nucleocapsid protein of PEDV[J].Chinese journal of preventive veterinary medicine,2008,(07):190.
[6]陈建飞,冯 力*,时洪艳,等.猪流行性腹泻病毒CH/S株N蛋白基因的遗传变异及其原核表达[J].中国预防兽医学报,2007,(11):856.
 CHEN Jian-fei,FENG Li*,SHI Hong-yan,et al.Genetic variation and prokaryotic expression of N protein gene of porcine epidemic diarrhea virus CH/S strain[J].Chinese journal of preventive veterinary medicine,2007,(07):856.
[7]孙东波,冯 力*,时洪艳,等.猪流行性腹泻病毒S蛋白中和表位区单克隆抗体的制备与鉴定[J].中国预防兽医学报,2007,(11):887.
 SUN Dong-bo,FENG Li*,SHI Hong-yan,et al.Preparation and characterization of the monoclonal antibodies against neutralizing epitope region of PEDV S protein[J].Chinese journal of preventive veterinary medicine,2007,(07):887.
[8]高慎阳,邹运明,李一经*.猪流行性腹泻病毒M基因及其片段原核表达效果分析[J].中国预防兽医学报,2007,(01):27.
 GAO Shen-yang,ZOU Yun-ming,LI Yi-jing*.Prokaryotic expression analysis of recombinant M gene and its truncated M’ fragment of porcine epidemic diarrhea virus[J].Chinese journal of preventive veterinary medicine,2007,(07):27.
[9]吴玉璐,朱建平,杨 莘,等.猪流行性腹泻病毒N基因的表达及抗原性分析[J].中国预防兽医学报,2013,(04):299.[doi:doi: 10.3969/j.issn.1008-0589.2013.04.11]
 WU Yu-lu,ZHU Jian-ping,YANG Shen,et al.Expression and antigenic analysis of porcine epidemic diarrhea virus N gene[J].Chinese journal of preventive veterinary medicine,2013,(07):299.[doi:doi: 10.3969/j.issn.1008-0589.2013.04.11]
[10]刘随新,石 达,陈建飞,等.猪流行性腹泻病毒N蛋白核仁定位信号对宿主细胞周期的影响[J].中国预防兽医学报,2013,(03):0.
 LIU Sui-xin,SHI Da,CHEN Jian-fei,et al.Effects on host cell cycle by nucleoli localization signal peptides of porcine epidemic diarrhea virus nucleocapsid protein[J].Chinese journal of preventive veterinary medicine,2013,(07):0.

备注/Memo

备注/Memo:
基金:国家十一五科技支撑计划资助项目(2006BAD06A07)
更新日期/Last Update: 2011-09-29