A Sensitive and High-Throughput Flow Cytometry-Based Assay for Measuring Antibody Neutralization of Human Adenovirus Type 3

作     者：Zhenwei Liu Xingui Tian Wenkuan Liu Yuting Xian Weilue Chen Rong Zhou Zhenwei Liu;Xingui Tian;Wenkuan Liu;Yuting Xian;Weilue Chen;Rong Zhou

作者机构：State Key Laboratory of Respiratory DiseaseNational Clinical Research Center for Respiratory DiseaseGuangzhou Institute of Respiratory HealthThe First Affiliated Hospital of Guangzhou Medical UniversityGuangzhou Medical UniversityGuangzhou 510000China 

出 版 物：《Virologica Sinica》 (中国病毒学（英文版）)

年 卷 期：2021年第36卷第3期

页      面：537-544页

核心收录：

学科分类：1010[医学-医学技术（可授医学、理学学位）] 10[医学] 

基　　金：supported by the National Science and Technology Major Project of China (2017ZX10103011-003, 2018ZX10102001) the National Natural Science Foundation of China (31570163) Guangzhou Science and Technology Program key projects (201803040004) 

主　　题：Sensitive High-throughput Flow cytometry Neutralization Adenovirus 

摘      要：The assessment of neutralization activity is an important step in the evaluation of neutralizing antibodies (NAbs).The traditional methods for measuring the antibody neutralization of human adenovirus type 3 (HAd V-3) are the microneutralization (MN) assay,which has insufficient sensitivity,and the plaque reduction neutralization test (PRNT),which is not suitable for high-throughput ***,we describe the development of a flow cytometry-based neutralization(FCN) assay for measuring the neutralization of sera,cell culture supernatants,and chimeric antibodies against HAd V-3 on the basis of a recombinant HAd V-3 (r HAd V-3) construct expressing the enhanced green fluorescent protein (EGFP).For flow cytometry-based assays,the optimal cell confluence was determined as 90%,and the virus was titrated using the *** established FCN assay follows the percentage law and an optimal MOI of not less than 5 9 10~(-4)was determined by using a purified chimeric *** addition,comparison of the anti-HAd V-3 NAb titers of 72 human serum samples by the MN and FCN assays,showed that both assays correlated strongly with each *** FCN assay was an improvement over the MN assay because the observation period was reduced from 3 to 1 days and data analysis could be performed objectively and ***,the newly established FCN assay allows measurement of the neutralization activity of chimeric antibodies expressed in cell culture ***,this sensitive and highthroughput FCN assay is a useful alternative to the MN assay for measuring the antibody neutralization of HAd V-3 and for screening anti-HAd V-3 NAbs in cell culture supernatants.