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.
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 149.36-157.23 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.
出处
《云南大学学报（自然科学版）》
CAS
CSCD
北大核心
2013年第1期93-98,共6页
Journal of Yunnan University(Natural Sciences Edition)
基金
中国烟草总公司云南省分公司科技计划项目(07A07)
作者简介
吕琦（1968-），女，云南人，实验师，主要从事抗体制备技术的研究开发．
通信作者：马岚（1965-），女，博士，研究员，主要从事细胞生物学及免疫学等研究．E-mail：roman@sz．tsinghua．edu．cn．
