猫传染性腹膜炎病毒S1蛋白的截短表达及其单克隆抗体的研制  

Truncated expression of S1 protein of feline infectious peritonitis virus and preparation of its monoclonal antibody

作　　者：赵辉 杜吉革[2] 李启红 勾倩倩[2] 黄金龙 刘妍 赵洋 杨海燕 印春生[2] 张传美[1] ZHAO Hui;DU Ji-ge;LI Qi-hong;GOU Qian-qian;HUANG Jin-long;LIU Yan;ZHAO Yang;YANG Hai-yan;YIN Chun-sheng;ZHANG Chuan-mei(College of Veterinary Medicine,Qingdao Agricultural University,Qingdao 266109,China;China Institute of Veterinary Drug Control,Beijing 100081,China)

机构地区：[1]青岛农业大学动物医学院,山东青岛266109 [2]中国兽医药品监察所,北京100081

出　　处：《中国兽医科学》2022年第9期1137-1143,共7页Chinese Veterinary Science

基　　金：中国兽医药品监察所“兽药行业公益性重点专项”(GY202005)。

摘　　要：为制备猫传染性腹膜炎病毒(FIPV)S1蛋白的单克隆抗体,本研究通过大肠杆菌表达系统截短表达了FIPV S1蛋白(rS1)并进行纯化,以rS1免疫BALB/c小鼠,通过细胞融合和间接ELISA方法筛选到3株能稳定分泌针对rS1抗体的杂交瘤细胞株,分别命名为2D7、3D8、5G1。抗体亚型鉴定结果表明,2D7、5G1株抗体亚型为Ig G2a,3D8株抗体亚型为Ig G1,轻链均为κappa链;杂交瘤细胞染色体组型鉴定结果显示,3株杂交瘤细胞染色体数分别为102、103、100条,属于杂交瘤染色体组型;小鼠腹水的间接ELISA抗体效价可达1∶204800。对小鼠腹水进行单克隆抗体纯化和鉴定,Western-blot结果显示,3株单克隆抗体均可与rS1发生特异性反应;IFA结果显示,3株单克隆抗体均能特异性识别细胞中的FIPV。上述结果表明,本研究获得了3株性能良好的抗rS1的单克隆抗体,为后期FIPV的早期诊断以及FIPV生物学功能的研究奠定了重要的物质基础。In order to develop monoclonal antibody against Feline infectious peritonitis virus(FIPV)S1 protein,the truncated S1 protein(r S1)was expressed through Escherichia coli and subsequently purified.Then BALB/c mice were immunized with purified r S1.Three hybridoma cell strains,named 2D7,3D8and 5G1,stably secreting antibodies against r S1 were obtained by cell fusion and indirect ELISA screening.The identification of antibody subtype showed that antibody subtypes of 2D7,5G1 and 3d8 strains were Ig G2a,Ig G2a and Ig G1,respectively.And the light chain of those three hybridoma cell strains wasκappa.Result of karyotype identification of hybridoma cells showed that the chromosome numbers of those three hybridoma cells were about 102,101 and 103,which was belonged to the karyotype of hybridoma.The titer of ascites antibody for indirect ELISA was 1∶204800,and monoclonal antibodies were purified.Moreover,all of 2D7,3D8 and 5G1 could react with r S1 by Western-blot and FIPV in cells by IFA.These data suggest that three monoclonal antibodies against r S1 with good activities were ideal materials in the study of early diagnosis of FIPV and the biological function of FIPV in the future.

关 键 词：猫传染性腹膜炎病毒 S1蛋白 原核表达 单克隆抗体 

分 类 号：S852.659.6[农业科学—基础兽医学]