| 烟草花叶病毒单克隆抗体的制备和鉴定 |
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| 引用本文： | 吕琦,吴峰,林洁,谭仲夏,秦西云,夏振远,莫笑晗,马岚. 烟草花叶病毒单克隆抗体的制备和鉴定[J]. 云南大学学报(自然科学版), 2013, 35(1): 93-98. DOI: 10.7540/j.ynu.2012.12072 |
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| 作者姓名： | 吕琦 吴峰 林洁 谭仲夏 秦西云 夏振远 莫笑晗 马岚 |
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| 作者单位： | 云南大学生命科学学院;清华大学深圳研究生院生命与健康学部;云南省烟草农业科学研究院 |
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| 基金项目： | 中国烟草总公司云南省分公司科技计划项目(07A07) |
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| 摘 要： | 经工程菌表达与纯化,得到了纯度95%以上的TMV-CP-F重组蛋白,配合提取的TMV天然病毒颗粒作为免疫原.通过杂交瘤技术获得了14株能分泌特异针对TMV外壳蛋白的单克隆抗体杂交瘤细胞株.经鉴定14株细胞所分泌的抗体亚类为IgG1型,抗体轻链均为κ型.经ProteinA一步法亲和层析纯化所得抗体经鉴定相对分子质量在149.36～157.23 ku之间,抗体纯度在80%以上.经间接ELISA测定,14株抗体均与TMV-CP重组蛋白和TMV病毒有良好特异性反应.所制备的抗TMV抗体的特异性高,可用于与其相关的免疫检测研究和应用.
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| 关 键 词： | 烟草花叶病毒 单克隆抗体 间接法ELISA |
Preparation and identification of monoclonal antibody against Tobacco mosaic virus |
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| LV Qi,WU Feng,LIN Jie,TAN Zhong-xia,QIN Xi-yun,XIA Zhen-yuan,MO Xiao-han,MA Lan. Preparation and identification of monoclonal antibody against Tobacco mosaic virus[J]. Journal of Yunnan University(Natural Sciences), 2013, 35(1): 93-98. DOI: 10.7540/j.ynu.2012.12072 |
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| Authors: | LV Qi WU Feng LIN Jie TAN Zhong-xia QIN Xi-yun XIA Zhen-yuan MO Xiao-han MA Lan |
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| Affiliation: | 1.School of Life Sciences,Yunnan University,Kunming 650091,China; 2.Division of Life Science & Health,Graduate School at Shenzhen,Tsinghua University,Shenzhen 518005,China; 3.Yunnan Academy of Tobacco Agricultural Sciences,Yuxi 653100,China) |
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| Abstract: | Recombinant TMV coat protein (TMV-CP) was expressed by engineered bacteria and then purified ( purity was above 95% ).It was used as immunogen together with the natural TMV particles for immunization.After hybridoma screening,14 hybridoma cell lines which stably secreted monoclonal antibodies against recombinant TMV-CP were obtained.The IgG subclasses and light-chain isotypes of all these antibodies were identified as IgG1 and κ respectively.The antibodies were purified by one-step recombinant protein-A affinity chromatography.Their molecular weight was among 149.36-157.23ku,and the purity was above 80%.The ELISA results showed that all of these 14 monoclonal antibodies reacted with recombinant TMV-CP and natural TMV particles specifically.The TMV monoclonal antibodies have high specificity and it can be used for relevant researches and immunoassays. |
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| Keywords: | Tobacco mosaic virus monoclonal antibody indirect ELISA |
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