Expression and Characterization of Recombinant, Tetrameric and Enzymatically Active Influenza Neuraminidase for the Setup of an Enzyme-Linked Lectin-Based Assay
| Autor: | Fabiola Giusti, Domenico Maione, Marua Prevato, Yasushi Uematsu, Alessandra Bonci, Roberta Cozzi, Ilaria Ferlenghi, Sylvie Bertholet, John L. Telford, Francois Legay, Giulia Anselmi, Ethan C. Settembre |
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| Rok vydání: | 2015 |
| Předmět: |
Enzyme-Linked Immunospot Assay
Swine Influenza vaccine Cross Protection Neuraminidase lcsh:Medicine Hemagglutinin (influenza) Hemagglutinin Glycoproteins Influenza Virus Cross Reactions Antibodies Viral medicine.disease_cause Antigenic drift Virus Cell Line law.invention Microbiology Birds Mice Viral Proteins Influenza A Virus H1N1 Subtype Orthomyxoviridae Infections law Lectins medicine Influenza A virus Animals lcsh:Science Antigens Viral Multidisciplinary Influenza A Virus H5N1 Subtype biology lcsh:R Virology Recombinant Proteins Influenza A virus subtype H5N1 Influenza Vaccines Influenza in Birds Antibody Formation biology.protein Recombinant DNA Female lcsh:Q Research Article |
| Zdroj: | PLoS ONE, Vol 10, Iss 8, p e0135474 (2015) PLoS ONE |
| ISSN: | 1932-6203 |
| DOI: | 10.1371/journal.pone.0135474 |
| Popis: | Developing a universal influenza vaccine that induces broad spectrum and longer-term immunity has become an important potentially achievable target in influenza vaccine research and development. Hemagglutinin (HA) and neuraminidase (NA) are the two major influenza virus antigens. Although antibody responses against influenza virus are mainly directed toward HA, NA is reported to be more genetically stable; hence NA-based vaccines have the potential to be effective for longer time periods. NA-specific immunity has been shown to limit the spread of influenza virus, thus reducing disease symptoms and providing cross-protection against heterosubtypic viruses in mouse challenge experiments. The production of large quantities of highly pure and stable NA could be beneficial for the development of new antivirals, subunit-based vaccines, and novel diagnostic tools. In this study, recombinant NA (rNA) was produced in mammalian cells at high levels from both swine A/California/07/2009 (H1N1) and avian A/turkey/Turkey/01/2005 (H5N1) influenza viruses. Biochemical, structural, and immunological characterizations revealed that the soluble rNAs produced are tetrameric, enzymatically active and immunogenic, and finally they represent good alternatives to conventionally used sources of NA in the Enzyme-Linked Lectin Assay (ELLA). |
| Databáze: | OpenAIRE |
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