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当前位置: 首页 > 文章 > 鸡新城疫病毒中和单克隆抗体的制备及鉴定 河南农业科学 2017,46 (1) 127-131
Position: Home > Articles > Production and Characterization of Neutralizing Monoclonal Antibodies against Newcastle Disease Virus Journal of Henan Agricultural Sciences 2017,46 (1) 127-131

鸡新城疫病毒中和单克隆抗体的制备及鉴定

作  者：
李青梅;王丽;冯丽丽;张雨杭;赵东;杨艳艳;邢广旭;柴书军;李赛赛;张丽萍;郭军庆;张改平
单  位：
河南省农业科学院动物免疫学重点实验室;河南省农业科学院农业经济与信息研究所;河南农业大学牧医工程学院
关键词：
鸡新城疫病毒;单克隆抗体;免疫过氧化物酶单层细胞试验;血凝抑制试验;中和试验;中和活性
摘  要：
采用差速离心法纯化鸡新城疫病毒(NDV)标准毒株F48E8,免疫Balb/c小鼠,应用杂交瘤细胞技术制备抗NDV单克隆抗体,旨在为NDV中和抗原表位分析奠定基础。将NDV感染BHK-21细胞,建立异源免疫过氧化物酶单层细胞试验(IPMA)的单克隆抗体检测方法,筛选获得14株分泌抗NDV单克隆抗体杂交瘤细胞株,其单克隆抗体腹水IPMA效价均在0.50×10-2~2.56×10-5(F48E8株和La Sota株)。血凝抑制试验(HI)结果表明,单克隆抗体1G6、2C1、4D2、5F2和13A5具有血凝抑制活性,其HI效价在(6~12)log2(F48E8株)和(9~11)log2(La Sota株)。病毒中和试验结果表明,单克隆抗体5F2和13A5对F48E8株和La Sota株均有明显的病毒中和活性,中和效价分别为1∶400~1∶800和1∶25。夹心ELISA结果表明,单克隆抗体5F2识别NDV HN蛋白,13A5识别F蛋白。综上,成功制备了具有中和活性的NDV单克隆抗体(5F2和13A5)。
译  名：
Production and Characterization of Neutralizing Monoclonal Antibodies against Newcastle Disease Virus
作  者：
LI Qingmei;WANG Li;FENG Lili;ZHANG Yuhang;ZHAO Dong;YANG Yanyan;XING Guangxu;CHAI Shujun;LI Saisai;ZHANG Liping;GUO Junqing;ZHANG Gaiping;Key Laboratory of Animal Immunology,Henan Academy of Agricultural Sciences;Institute of Agricultural Economics and Information,Henan Academy of Agricultural Sciences;College of Animal Husbandry and Veterinary Medicine,Henan Agricultural University;Jiangsu Co-innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses;
关键词：
Newcastle disease virus;;monoclonal antibodies;;immunoperoxidase monolayer assay;;hemagglutination inhibition assay;;neutralization assay;;neutralization activity
摘  要：
Balb/c mice were immunized by the Newcastle disease virus( NDV) reference strain F48E8 purified by differential centrifugation,and used for monoclonal antibodies( m Abs) production using hybridoma technology. After screened by a heterologous immunoperoxidase monolayer assay( IPMA) based on the NDV-infected BHK-21 cells,fourteen hybridoma cell lines secreting m Abs specific for NDV were obtained,whose antibody titers of ascites were determined to be 0. 50 × 10-2—2. 56 × 10-5to F48E8 and toLa Sotain by IPMA respectively. In hemagglutination inhibition( HI) assay,m Abs 1G6,2C1,4D2,5F2 and13A5 showed hemagglutination-inhibitory activity with the HI titers of 6—12log2( F48E8) and 9—11log2( La Sota),of which m Abs 5F2 and 13A5 showed significant neutralizing activity to both NDV virulent and vaccine strains in the virus neutralization( VN) test with the titers of 1∶ 400—1∶ 800 and 1∶ 25 respectively. Furthermore,sandwich ELISA using the recombinant proteins showed that m Ab 5F2 reacted with HN protein of NDV,while m Ab 13A5 recognized the F protein. In conclusion,the NDV m Abs of 5F2 and13A5 with neutralization activity were obtained successfully.

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