河南农业科学2017，Vol.46Issue(1)：127-131,5.DOI:10.15933/j.cnki.1004-3268.2017.01.023
鸡新城疫病毒中和单克隆抗体的制备及鉴定
Production and Characterization of Neutralizing Monoclonal Antibodies against Newcastle Disease Virus
摘要
Abstract
Balb/c mice were immunized by the Newcastle disease virus (NDV) reference strain F48E8 purified by differential centrifugation,and used for monoclonal antibodies(mAbs) 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 mAbs specific for NDV were obtained,whose antibody titers of ascites were determined to be 0.50 × 10-2-2.56 × 10-5 to F48E8 and to LaSotain by IPMA respectively.In hemagglutination inhibition(HI) assay,mAbs 1 G6,2C1,4D2,5F2 and 13A5 showed hemagglutination-inhibitory activity with the HI titers of 6-12log2(F48E8) and 9-111og2 (LaSota),of which mAbs 5F2 and 13 A5 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 respective-ly.Furthermore,sandwich ELISA using the recombinant proteins showed that mAb 5F2 reacted with HN protein of NDV,while mAb 13A5 recognized the F protein.In conclusion,the NDV mAbs of 5F2 and 13A5 with neutralization activity were obtained successfully.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Key words
Newcastle disease virus/monoclonal antibodies/immunoperoxidase monolayer assay/hemagglutination inhibition assay/neutralization assay/neutralization activity分类
农业科技引用本文复制引用
李青梅,张丽萍,郭军庆,张改平,王丽,冯丽丽,张雨杭,赵东,杨艳艳,邢广旭,柴书军,李赛赛..鸡新城疫病毒中和单克隆抗体的制备及鉴定[J].河南农业科学,2017,46(1):127-131,5.基金项目
国家重点研发计划项目(2016YFD0500800) （2016YFD0500800）
公益性行业(农业)科研专项(201303033) （农业）
河南省科技攻关项目(162102110051) （162102110051）
