Development of a high-throughput assay to detect antibody inhibition of low pH induced conformational changes of influenza virus hemagglutinin

被引：4

作者：

Trost, Jessica F. ^{[1
,2
]}

LeMasters, Elizabeth H. ^{[1
]}

Liu, Feng ^{[1
]}

Carney, Paul ^{[1
]}

Lu, Xiuhua ^{[1
]}

Sugawara, Kanetsu ^{[3
]}

Hongo, Seiji ^{[3
]}

Stevens, James ^{[1
]}

Steinhauer, David A. ^{[2
]}

Tumpey, Terrence ^{[1
]}

Katz, Jacqueline M. ^{[1
]}

Levine, Min Z. ^{[1
]}

Li, Zhu-Nan ^{[1
]}

机构：

[1] Ctr Dis Control & Prevent, Influenza Div, Atlanta, GA 30333 USA

[2] Emory Univ, Sch Med, Dept Microbiol & Immunol, Atlanta, GA 30322 USA

[3] Yamagata Univ, Fac Med, Dept Infect Dis, Yamagata, Japan

来源：

PLOS ONE | 2018年 / 13卷 / 06期

基金：

美国国家卫生研究院;

关键词：

BROADLY NEUTRALIZING ANTIBODIES; MONOCLONAL-ANTIBODIES; STEM; PROTEINS; A(H3N2); EPITOPE; IDENTIFICATION; OPTIMUM; DOMAIN; SITES;

D O I：

10.1371/journal.pone.0199683

中图分类号：

O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];

学科分类号：

07 ; 0710 ; 09 ;

摘要：

Many broadly neutralizing antibodies (bnAbs) bind to conserved areas of the hemagglutinin (HA) stalk region and can inhibit the low pH induced HA conformational changes necessary for viral membrane fusion activity. We developed and evaluated a high-throughput virus-free and cell-free ELISA based low pH induced HA Conformational Change Inhibition Antibody Detection Assay (HCCIA) and a complementary proteinase susceptibility assay. Human serum samples (n = 150) were tested by HCCIA using H3 recombinant HA. Optical density (OD) ratios of mAb HC31 at pH 4.8 to pH 7.0 ranged from 0.87 to 0.09. Our results demonstrated that low pH induced HA conformational change inhibition antibodies (CCI) neutralized multiple H3 strains after removal of head-binding antibodies. The results suggest that HCCIA can be utilized to detect and characterize CCI in sera, that are potentially broadly neutralizing, and serves as a useful tool for evaluating universal vaccine candidates targeting the HA stalk.

引用

页数：21

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