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
引用本文: 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
shu
Preparation and identification of monoclonal antibodyagainst Tobacco mosaic virus[J]. Journal of Yunnan University: Natural Sciences Edition, 2013, 35(1): 93-98. DOI: 10.7540/j.ynu.2012.12072
Citation: Preparation and identification of monoclonal antibodyagainst Tobacco mosaic virus[J]. Journal of Yunnan University: Natural Sciences Edition, 2013, 35(1): 93-98. DOI: 10.7540/j.ynu.2012.12072
shu

烟草花叶病毒单克隆抗体的制备和鉴定

Preparation and identification of monoclonal antibodyagainst Tobacco mosaic virus

    摘要
  • 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. 

     

    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 respectively.The antibodies were purified by one-step recombinant protein-A affinity chromatography.Their molecular weight was among 149.36-157.23ku,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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