Expression and Characterization of Recombinant, Tetrameric and Enzymatically Active Influenza Neuraminidase for the Setup of an Enzyme-Linked Lectin-Based Assay.
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).
Research Center, Novartis Vaccines and Diagnostics s.r.l., (a GSK Company), Siena, Italy.
Marua
M
Prevato
Research Center, Novartis Vaccines and Diagnostics s.r.l., (a GSK Company), Siena, Italy.
Ilaria
I
Ferlenghi
Research Center, Novartis Vaccines and Diagnostics s.r.l., (a GSK Company), Siena, Italy.
Alessandra
A
Bonci
Research Center, Novartis Vaccines and Diagnostics s.r.l., (a GSK Company), Siena, Italy.
Yasushi
Y
Uematsu
Research Center, Novartis Vaccines and Diagnostics s.r.l., (a GSK Company), Siena, Italy.
Giulia
G
Anselmi
Department of Life Sciences, University of Siena, Siena, Italy.
Fabiola
F
Giusti
Research Center, Novartis Vaccines and Diagnostics s.r.l., (a GSK Company), Siena, Italy.
Sylvie
S
Bertholet
Vaccine Research, Novartis Vaccines and Diagnostics, (a GSK Company), Basel, Switzerland.
Francois
F
Legay
Research Center, Novartis Vaccines and Diagnostics s.r.l., (a GSK Company), Siena, Italy.
John Laird
JL
Telford
Vaccine Research, Novartis Vaccines and Diagnostics Inc., (a GSK Company), Cambridge, MA, United States of America.
Ethan C
EC
Settembre
Research Center, Novartis Vaccines and Diagnostics s.r.l., (a GSK Company), Siena, Italy.
Domenico
D
Maione
Research Center, Novartis Vaccines and Diagnostics s.r.l., (a GSK Company), Siena, Italy.
Roberta
R
Cozzi
D000818:Animals; D000914:Antibodies, Viral / Q000276:immunology; D000917:Antibody Formation / Q000276:immunology; D000956:Antigens, Viral / Q000276:immunology*; D001717:Birds; D002460:Cell Line; D056738:Cross Protection / Q000276:immunology; D003429:Cross Reactions / Q000276:immunology; D058501:Enzyme-Linked Immunospot Assay / Q000379:methods; D005260:Female; D019267:Hemagglutinin Glycoproteins, Influenza Virus / Q000276:immunology; D053118:Influenza A Virus, H1N1 Subtype / Q000276:immunology; D053124:Influenza A Virus, H5N1 Subtype / Q000276:immunology; D007252:Influenza Vaccines / Q000276:immunology; D005585:Influenza in Birds / Q000276:immunology; D037102:Lectins / Q000276:immunology*; D051379:Mice; D009439:Neuraminidase / Q000276:immunology*; D009976:Orthomyxoviridae Infections / Q000276:immunology; D011994:Recombinant Proteins / Q000276:immunology*; D013552:Swine; D014764:Viral Proteins / Q000276:immunology*
D000914:Antibodies, Viral; D000956:Antigens, Viral; D019267:Hemagglutinin Glycoproteins, Influenza Virus; D007252:Influenza Vaccines; D037102:Lectins; D011994:Recombinant Proteins; D014764:Viral Proteins; D009439:Neuraminidase