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| 口蹄疫病毒非结构蛋白3B单克隆抗体的制备与鉴定 | |
| 引用本文： | 李永亮,田美娜,卢曾军,杨苏珍,付元芳,曹轶梅,刘在新.口蹄疫病毒非结构蛋白3B单克隆抗体的制备与鉴定[J].江苏农业学报,2009,25(2). |
| 作者姓名： | 李永亮 田美娜 卢曾军 杨苏珍 付元芳 曹轶梅 刘在新 |
| 作者单位： | 1. 中国农业科学院兰州兽医研究所,国家口蹄疫参考实验室,家畜疫病病原生物学国家重点实验室,甘肃兰州730046 2. 河南省农业科学院动物免疫学重点实验室,河南郑州,450002 3. 甘肃农业大学动物医学院,甘肃兰州,730070 |
| 基金项目： | 国家重点基础研究发展规划(973计划) |
| 摘 要： | 用E.coli原核表达并纯化的口蹄疫病毒(Foot-and-mouth disease virus, FMDV)非结构蛋白3B免疫BALB/c小鼠,取其脾细胞与小鼠骨髓瘤细胞SP2/0进行融合,间接ELISA筛选出分泌鼠IgG的杂交瘤细胞株,将该杂交瘤细胞注射小鼠产生的腹水,用间接ELISA法筛选获得6株能稳定分泌抗3B蛋白单克隆抗体的杂交瘤细胞株,分别命名为3E5、4B1、4D7、4E11、7B2、8B11.鉴定结果显示,4B1和4E11细胞分泌IgG1,其余4株细胞分泌IgG2b;纯化后6株腹水单抗的纯度达90%以上,对3B蛋白的ELISA滴度均可达到1:100 000以上;6株单抗均不与FMDV结构蛋白VP1和3D非结构蛋白发生反应;间接免疫荧光试验证明所制备的单抗能够识别3B蛋白;杂交瘤细胞株连续培养3个月以及冻存6个月后复苏,细胞生长良好,杂交瘤细胞分泌的抗体效价稳定. |
| 关 键 词： | 口蹄疫病毒 非结构蛋白 3B蛋白 单克隆抗体 |
Preparation and Identification of Monoclonal Antibody (MAb) against Non-structural Protein 3B of Foot-and-mouth Disease |
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| LI Yong-liang,TIAN Mei-na,LU Zeng-jun,YANG Su-zhen,FU Yuan-fang,CAO Yi-mei,LIU Zai-xin.Preparation and Identification of Monoclonal Antibody (MAb) against Non-structural Protein 3B of Foot-and-mouth Disease[J].Jiangsu Journal of Agricultural Sciences,2009,25(2). | |
| Authors: | LI Yong-liang TIAN Mei-na LU Zeng-jun YANG Su-zhen FU Yuan-fang CAO Yi-mei LIU Zai-xin |
| Institution: | 1.State Key Laboratory of Veterinary Etiological Biology;National Foot-and-Mouth Disease Reference Laboratory;Lanzhou Veterinary Research Institute;Chinese Academy of Agricultural Sciences;Lanzhou 730046;China;2.Key Laboratory for Animal Immunology;Henan Academy of Agricultural Sciences;Zhengzhou 450002;3.Veterinary College;Gansu Agricultural University;Lanzhou 730070;China |
| Abstract: | The monoclonal antibodies against protein 3B of foot-and-mouth disease virus were generated by fusing spleen cells from 3B immunized BALB/c mouse with SP2/0 mouse myeloma cells.The culture of hybridoma was screened by I-ELISA for detection of specific mouse IgG against FMDV 3B protein.Six strains of hybridoma which secreted monoclonal antibodies against protein 3B were determined and named 3E5,4B1,4D7,4E11,7B2 and 8B11.The isotypes were identified to be IgG1 and IgG2b.The titer of these MAbs were over 105 b... |
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