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| H9亚型禽流感病毒基质蛋白M1单克隆抗体的制备与鉴定 | |
| 引用本文： | 戴晓光,滕巧泱,黄海碧,王朝霞,张树梅,张旭,徐大伟,申之义,李泽君. H9亚型禽流感病毒基质蛋白M1单克隆抗体的制备与鉴定[J]. 中国兽医寄生虫病, 2010, 18(3): 8-12 |
| 作者姓名： | 戴晓光 滕巧泱 黄海碧 王朝霞 张树梅 张旭 徐大伟 申之义 李泽君 |
| 作者单位： | 1. 内蒙古农业大学兽医学院,呼和浩特010018;中国农业科学院上海兽医研究所,农业部动物寄生虫学重点开放实验室,上海,200241 2. 中国农业科学院上海兽医研究所,农业部动物寄生虫学重点开放实验室,上海,200241 3. 内蒙古农业大学兽医学院,呼和浩特,010018 |
| 摘 要： | 本研究用纯化的H9N2亚型禽流感病毒免疫BALB/c小鼠,取其脾细胞与SP2/0骨髓瘤细胞融合。对杂交瘤细胞及时筛选,阳性孔经3次有限稀释法克隆,成功获得3株能稳定传代并分泌抗H9亚型禽流感病毒基质蛋白M1单克隆抗体的杂交瘤细胞：3G8、2F6、5F2。间接ELISA方法检测,3株单克隆抗体的腹水间接ELISA效价达106以上。构建了真核表达载体pCAGGS-M1并转染于MDCK细胞,以用于腹水的Western blot和间接免疫荧光鉴定。间接免疫荧光结果表明,3株单克隆抗体皆与真核表达蛋白M1反应。这些单克隆抗体的制备为后期研究M1蛋白在流感病毒复制与出芽过程中的重要生物学功能奠定了基础。 |
| 关 键 词： | H9亚型禽流感病毒 基质蛋白M1 单克隆抗体 间接免疫荧光 蛋白免疫印迹法 |
PREPARATION AND CHARACTERIZATION OF MONOCLONAL ANTIBODIES AGAINST M1 PROTEIN OF H9N2 AVIAN INFLUENZA VIRUS |
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| DAI Xiao-guang,TENG Qiao-yang,HUANG Hai-bi,WANG Zhao-xia,ZHANG Shu-mei,ZHANG Xu,XU Da-wei,SHEN Zhi-yi,LI Ze-jun. PREPARATION AND CHARACTERIZATION OF MONOCLONAL ANTIBODIES AGAINST M1 PROTEIN OF H9N2 AVIAN INFLUENZA VIRUS[J]. Chinese Journal of Veterinary Parasitology, 2010, 18(3): 8-12 | |
| Authors: | DAI Xiao-guang TENG Qiao-yang HUANG Hai-bi WANG Zhao-xia ZHANG Shu-mei ZHANG Xu XU Da-wei SHEN Zhi-yi LI Ze-jun |
| Affiliation: | 1.Veterinary School of Inner Mongolia Agricultural University,Hohhot 010018,China;2.Key Laboratory of Animal Parasitology of Ministry of Agriculture,Shanghai Veterinary Research Institute,CAAS,Shanghai 200241,China) |
| Abstract: | BALB/c mice were immunized with the H9N2 subtype avian influenza virus and their spleen cells were fused with mouse myeloma cells（SP2/0）.Three stable hybridoma cell lines（3G8,2F6,5F2） secreting monoclonal antibodies（mAbs） against matrix protein M1 were identified after three rounds of limited dilutions.The titers of ascetic fluids of these three mAbs were up to 106 by indirect ELISA.In order to test the mAbs in Western blot and indirect inmmunofluorescene assay（IFA）,the recombinant eukaryotic expression vector pCAGGS-M1 was constructed and transfected into MDCK cells.The results of IFA demonstrated that MAbs could specifically bind to eukaryotic M1 protein.However,these MAbs could not react with M1 protein in Western blots,suggesting they might be specific for structural epitopes of M1.These MAbs can be used to analyze biological function of M1 protein in the process of virus replication and budding. |
| Keywords: | Avian influenza virus matrix protein monoclonal antibody indirect inmmunofluorescene assay（IFA） Western blot |
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