A Carboxy-Terminal Trimerization Domain Stabilizes Conformational Epitopes on the Stalk Domain of Soluble Recombinant Hemagglutinin Substrates

被引：145

作者：

Krammer, Florian ^{[1
]}

Margine, Irina ^{[1
,3
]}

Tan, Gene S. ^{[1
]}

Pica, Natalie ^{[1
,3
]}

Krause, Jens C. ^{[1
]}

Palese, Peter ^{[1
,2
]}

机构：

[1] Mt Sinai Sch Med, Dept Microbiol, New York, NY USA

[2] Mt Sinai Sch Med, Dept Med, New York, NY USA

[3] Mt Sinai Sch Med, Grad Sch Biol Sci, New York, NY USA

来源：

PLOS ONE | 2012年 / 7卷 / 08期

基金：

奥地利科学基金会;

关键词：

INFLUENZA-A VIRUS; MONOCLONAL-ANTIBODIES; NEUTRALIZING EPITOPE; H1N1; EFFICACY; H5N1; CELLS;

D O I：

10.1371/journal.pone.0043603

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：

Recently, a new class of broadly neutralizing anti-influenza virus antibodies that target the stalk domain of the viral hemagglutinin was discovered. As such, induction, isolation, characterization, and quantification of these novel antibodies has become an area of intense research and great interest. Since most of these antibodies bind to conformational epitopes, the structural integrity of hemagglutinin substrates for the detection and quantification of these antibodies is of high importance. Here we evaluate the binding of these antibodies to soluble, secreted hemagglutinins with or without a carboxy-terminal trimerization domain based on the natural trimerization domain of T4 phage fibritin. The lack of such a domain completely abolishes binding to group 1 hemagglutinins and also affects binding to group 2 hemagglutinins. Additionally, the presence of a trimerization domain positively influences soluble hemagglutinin stability during expression and purification. Our findings suggest that a carboxy-terminal trimerization domain is a necessary requirement for the structural integrity of stalk epitopes on recombinant soluble influenza virus hemagglutinin.

引用

页数：10

共 34 条

[11] Bacterial HA1 Vaccine against Pandemic H5N1 Influenza Virus: Evidence of Oligomerization, Hemagglutination, and Cross-Protective Immunity in Ferrets [J].