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| 猪繁殖与呼吸综合征病毒GP5蛋白单克隆抗体的制备与鉴定 | |
| 引用本文： | 王志红,张改平,郭军庆,李青梅,王寅彪,陈稳,王爱萍.猪繁殖与呼吸综合征病毒GP5蛋白单克隆抗体的制备与鉴定[J].河南农业科学,2012,41(6):144-147. |
| 作者姓名： | 王志红 张改平 郭军庆 李青梅 王寅彪 陈稳 王爱萍 |
| 作者单位： | 1. 郑州大学生物工程系,河南郑州450001;河南省农业科学院农业部动物免疫学重点开放实验室/河南省动物免疫学重点实验室,河南郑州450002 2. 河南省农业科学院农业部动物免疫学重点开放实验室/河南省动物免疫学重点实验室,河南郑州,450002 3. 郑州大学生物工程系,河南郑州,450001 |
| 基金项目： | 国家自然科学基金面上项目(31072122) |
| 摘 要： | 为了制备猪繁殖与呼吸综合征病毒(PPRSV)的单克隆抗体,利用猪繁殖与呼吸综合征病毒(PPRSV)GP5重组蛋白免疫BALB/c小鼠,取脾细胞和NS0浆细胞瘤细胞进行融合,经间接ELISA和阻断ELISA筛选、有限稀释法克隆化获得3株能稳定分泌抗PRRSV GP5蛋白单克隆抗体的(mAb)杂交瘤细胞株,命名为4B7、2C5和2E4。间接免疫荧光(IFA)和免疫过氧化物酶单层试验(IPMA)结果显示,3株mAb均与感染PRRSV BJ 4株的Marc 145细胞特异性反应,表明其可识别天然病毒蛋白。mAb的Ig亚类均为IgG1,细胞培养上清和腹水的ELISA效价分别为1∶256和1∶51 200,Western blot分析表明,mAb均特异性识别PRRSV GP5蛋白。 |
| 关 键 词： | 猪繁殖与呼吸综合征病毒 GP5蛋白 单克隆抗体 |
Development and Characterization of Monoclonal Antibody against GP5 Protein of Porcine Reproduction and Respiratory Syndrome Virus |
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| WANG Zhi-hong , ZHANG Gai-ping , GUO Jun-qing , LI Qing-mei , WANG Yin-biao , CHEN Wen , WANG Ai-ping.Development and Characterization of Monoclonal Antibody against GP5 Protein of Porcine Reproduction and Respiratory Syndrome Virus[J].Journal of Henan Agricultural Sciences,2012,41(6):144-147. | |
| Authors: | WANG Zhi-hong ZHANG Gai-ping GUO Jun-qing LI Qing-mei WANG Yin-biao CHEN Wen WANG Ai-ping |
| Institution: | 1 (1.Department of Bioengineering,Zhengzhou University,Zhengzhou 450001,China; 2.Key Laboratory of Animal Immunology of the Ministry of Agriculture/Henan Provincial Key Laboratory of Animal Immunology,Henan Academy of Agriculture Sciences,Zhengzhou 450002,China) |
| Abstract: | In order to prepare monoclonal antibodies(mAbs) against porcine reproductive and respiratory syndrome virus(PPRSV),the recombination GP5 protein of PRRSV was used to immunize BALB/c mouse and the spleen cells were fused with NS0 cells.Three hybridoma cell lines producing mAbs specific to the GP5 protein,designated as 4B7,2C5 and 2E4,were obtained after identified by indirect and inhibit ELISAs followed by limiting dilution subcloning.All of the mAbs were shown to react specifically with the Marc-145 cells infected with PRRSV BJ-4 strain in indirect immunofluorescence assay(IFA) andimmunoperoxidase monolayer assays(IPMA).Ig subclass of the mAbs was determined as IgG1,and the ELISA titers of cell supernatant and ascites were 1∶256 and 1∶51 200,respectively.Western-blotanalysis showed that the mAbs were specific to the GP5 protein of PRRSV. |
| Keywords: | porcine reproduction and respiratory syndrome virus(PRRSV) GP5 protein monoclonal antibody |
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