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| 应用假病毒和假型病毒制备抗猪瘟病毒E2蛋白中和性单克隆抗体的初步研究 | |
| 引用本文： | 陈景艳,尹晓磊,王宜,袁丽,王超,邓宏魁,钱永华,袁克湖. 应用假病毒和假型病毒制备抗猪瘟病毒E2蛋白中和性单克隆抗体的初步研究[J]. 中国预防兽医学报, 2010, 32(4) |
| 作者姓名： | 陈景艳 尹晓磊 王宜 袁丽 王超 邓宏魁 钱永华 袁克湖 |
| 作者单位： | 1. 西北农林科技大学,动物医学院,陕西,杨凌,712100 2. 北京大学,深圳研究生院,广东,深圳,518055;北京大学,生命科学学院,北京,100871 3. 北京大学,深圳研究生院,广东,深圳,518055 |
| 摘 要： | 为制备抗猪瘟病毒(CSFV)单克隆抗体(MAb),本实验以表达CSFV E2囊膜糖蛋白的水泡口炎假病毒免疫BALB/c小鼠,取其脾细胞与骨髓瘤细胞SP2/0进行融合;利用间接ELISA方法和携带荧光素酶(Luciferase)报告基因的HIV-luc/CSFV-E1E2假病毒系统筛选分泌中和性E2MAb的杂交瘤细胞;测定MAb亚型并纯化后,间接ELISA方法测定MAb的效价;采用western blot鉴定MAb的特异性亲和力;利用HIV-luc/CSFV-E1E2假病毒进行体外中和试验,分析MAb抑制病毒感染的能力。结果表明本实验获得了1株分泌中和性MAb的杂交瘤细胞9C8,该MAb能与E2蛋白特异性结合,而且体外抑制试验中和效价大于1∶25600。 |
| 关 键 词： | 猪瘟病毒 中和性单克隆抗体 假病毒 中和试验 |
Development of neutralizing monoclonal antibody against E2 protein of CSFV by using pseudovirus and pseudotyped virus |
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| CHEN Jing-yan,YIN Xiao-lei,WANG Yi,YUAN Li,WANG Chao,DENG Hong-kui,QIAN Yong-hua,YUAN Ke-hu. Development of neutralizing monoclonal antibody against E2 protein of CSFV by using pseudovirus and pseudotyped virus[J]. Chinese Journal of Preventive Veterinary Medicine, 2010, 32(4) | |
| Authors: | CHEN Jing-yan YIN Xiao-lei WANG Yi YUAN Li WANG Chao DENG Hong-kui QIAN Yong-hua YUAN Ke-hu |
| Affiliation: | CHEN Jing-yan1,YIN Xiao-lei2,3,WANG Yi2,YUAN Li2,WANG Chao2,DENG Hong-kui2,QIAN Yong-hua1,YUAN Ke-hu2(1. College of Animal Science,Northwest Agriculture , Forestry University,Yangling 712100,China,2. Shenzhen Graduate School of Peking University,Shenzhen 518055,3. College of Bioscience,Peking University,Beijing 100871,China) |
| Abstract: | To prepare E2 neutralizing monoclonal antibody(MAb) against classical swine fever virus(CSFV) ,BALB/c mice were immunized with recombinant vesicular stomatitis virus(rVSV) expressing the envelope glycoprotein E2 of CSFV;mouse splenic cells were fused with SP2/0 cells and hybridoma cells were screened by indirect ELISA and retroviral pseudotype virus HIV-luc/CSFV bearing CSFV E1 and E2 glycoproteins. The specificity and the antigen-binding activity of the MAb were identified by indirect ELISA and western blo... |
| Keywords: | CSFV neutralizing monocolonal antibody pseudotyped virus neutralization assay |
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