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抗狂犬病病毒单克隆抗体的制备与鉴定。

Preparation and identification of anti-rabies virus monoclonal antibodies.

机构信息

College of Animal Science and Veterinary Medicine, Qingdao Agricultural university, Qingdao 266109, China.

出版信息

Virol Sin. 2012 Jun;27(3):172-8. doi: 10.1007/s12250-012-3242-0. Epub 2012 Jun 9.

Abstract

To provide a foundation for the development of rapid and specific methods for the diagnosis of rabies virus infection, anti-rabies virus monoclonal antibodies were prepared and rabies virus nucleoprotein and human rabies virus vaccine strain (PV strain) were used as immunogens to immunize 6-8 week old female BALB/c mice. Spleen cells and SP2/0 myeloma cells were fused according to conventional methods: the monoclonal cell strains obtained were selected using the indirect immunofluorescence test; this was followed by preparation of monoclonal antibody ascitic fluid; and finally, systematic identification of subclass, specificity and sensitivity was carried out. Two high potency and specific monoclonal antibodies against rabies virus were obtained and named 3B12 and 4A12, with ascitic fluid titers of 1:8000 and 1:10000, respectively. Both belonged to the IgG2a subclass. These strains secrete potent, stable and specific anti-rabies virus monoclonal antibodies, which makes them well suited for the development of rabies diagnosis reagents.

摘要

为了为快速和特异性诊断狂犬病病毒感染的方法的发展提供基础,制备了抗狂犬病病毒单克隆抗体,并用人狂犬病病毒核蛋白和狂犬病病毒疫苗株(PV 株)作为免疫原,免疫 6-8 周龄雌性 BALB/c 小鼠。根据常规方法使脾细胞和 SP2/0 骨髓瘤细胞融合:使用间接免疫荧光试验选择获得的单克隆细胞株;然后制备单克隆抗体腹水;最后,进行了亚类、特异性和敏感性的系统鉴定。获得了两种针对狂犬病病毒的高效价和特异性单克隆抗体,分别命名为 3B12 和 4A12,腹水效价分别为 1:8000 和 1:10000,均属于 IgG2a 亚类。这些株系分泌强效、稳定和特异性的抗狂犬病病毒单克隆抗体,非常适合开发狂犬病诊断试剂。

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