A carboxy-terminal trimerization domain stabilizes conformational epitopes on the stalk domain of soluble recombinant hemagglutinin substrates

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 interes...

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Bibliographic Details
Published in:PloS one 2012-08, Vol.7 (8), p.e43603-e43603
Main Authors: Krammer, Florian, Margine, Irina, Tan, Gene S, Pica, Natalie, Krause, Jens C, Palese, Peter
Format: Article
Language:English
Subjects:
Animals
Antibiotics
Antibodies
Antibodies, Viral - immunology
Avian flu
Binding
Biology
Epitopes
Epitopes - chemistry
Epitopes - immunology
Freezing
Gene expression
Hemagglutinin Glycoproteins, Influenza Virus - chemistry
Hemagglutinin Glycoproteins, Influenza Virus - immunology
Hemagglutinins
Humans
Immunoglobulins
Influenza
Influenza A virus - immunology
Influenza viruses
Lectins
Medicine
Monoclonal antibodies
Orthomyxoviridae
Pandemics
Penicillin
Phages
Protein Multimerization
Protein Stability
Protein Structure, Tertiary
Purification
Recombinant Proteins - chemistry
Recombinant Proteins - immunology
Sf9 Cells
Solubility
Structural integrity
Substrates
Swine flu
Vaccines
Viruses
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Summary: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.
ISSN:1932-6203
1932-6203
DOI:10.1371/journal.pone.0043603