| H5N1禽流感病毒PB1-F2特异性单克隆抗体的制备与鉴定 |
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| 引用本文： | 徐国双,刘恩华,张茂林,段铭,关振宏.H5N1禽流感病毒PB1-F2特异性单克隆抗体的制备与鉴定[J].动物医学进展,2020(7):1-6. |
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| 作者姓名： | 徐国双 刘恩华 张茂林 段铭 关振宏 |
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| 作者单位： | 吉林大学人兽共患病教育部重点实验室/人兽共患病研究所 |
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| 基金项目： | 吉林省科学技术厅自然科学基金项目(20160101214JC)。 |
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| 摘 要： | 以重组的H5N1亚型禽流感病毒(AIV)PB1-F2的C端蛋白(cPB1-F2)为免疫原制备单克隆抗体,并鉴定其特异性。构建cPB1-F2的pGEX-6p-1原核表达载体,进行蛋白表达与纯化;采用杂交瘤技术制备筛选抗重组cPB1-F2蛋白的杂交瘤细胞株,利用间接免疫荧光和酶联免疫吸附试验筛选鉴定阳性细胞株,并对单克隆抗体特异性进行鉴定。结果显示,纯化的重组cPB1-F2蛋白具有良好的免疫原性,筛选获得了3株稳定分泌抗cPB1-F2单克隆抗体的杂交瘤细胞株,分别命名为6F4、2C5和4D3,经鉴定抗体均为IgG1亚类,且3株单克隆抗体均能特异性识别重组PB1-F2蛋白。结果表明成功制备了cPB1-F2特异性单克隆抗体,为研制针对流感病毒的抗体药物奠定了基础。
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| 关 键 词： | 禽流感病毒 H5N1 PB1-F2 单克隆抗体 |
Preparation and Identification of Monoclonal Antibodies against PB1-F2 Protein of H5N1 Avian Influenza Virus |
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| XU Guo-shuang,LIU En-hua,ZHANG Mao-lin,DUAN Ming,GUAN Zhen-hong.Preparation and Identification of Monoclonal Antibodies against PB1-F2 Protein of H5N1 Avian Influenza Virus[J].Progress In Veterinary Medicine,2020(7):1-6. |
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| Authors: | XU Guo-shuang LIU En-hua ZHANG Mao-lin DUAN Ming GUAN Zhen-hong |
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| Institution: | (Key Laboratory of Zoonosis Ministry of Education,Institute of Zoonosis,Jilin University,Changchun,Jilin,130062,China) |
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| Abstract: | To prepare monoclonal antibodies against H5N1 subtype avian influenza virus(AIV)PB1-F2 using the recombinant C-terminal PB1-F2(cPB1-F2)as an immunogen,and further to identify their specificity,the pGEX-6p-1 prokaryotic expression vector of cPB1-F2 was constructed,and then the recombinant protein was expressed and purified.The anti-cPB1-F2 hybridoma cell lines were screened and produced by using hybridoma techniques,and positive cell strains were determined using indirect immunofluorescence assay(IFA)and enzyme-linked immunosorbent assay(ELISA).Finally,three strains of hybridoma cell lines secreting monoclonal antibodies against cPB1-F2 were established,named 6F4,2C5 and 4D3 respectively,and were all identified as IgG1 subtype.And three of them could specifically recognize recombinant PB1-F2 protein.In this study,the specific monoclonal antibodies against AIV(H5N1)cPB1-F2 were successfully screened and prepared,which laid a foundation for the development of antibody drugs for prevention and treatment of influenza virus. |
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| Keywords: | Avian influenza virus H5N1 PB1-F2 monoclonal antibody |
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