title: Generation and characterization of a new panel of broadly reactive anti-NS1 mAbs for detection of influenza A virus.
abstract: Influenza A virus (IAV) non-structural protein 1 (NS1) has multiple functions, is essential for virus replication and may be a good target for IAV diagnosis. To generate broadly cross-reactive NS1-specific mAbs, mice were immunized with A/Hong Kong/1/1968 (H3N2) 6×His-tagged NS1 and hybridomas were screened with glutathione S-transferase-conjugated NS1 of A/Puerto Rico/8/1934 (H1N1). mAbs were isotyped and numerous IgG-type clones were characterized further. Most clones specifically recognized NS1 from various H1N1 and H3N2 IAV types by both immunoblot and immunofluorescence microscopy in mouse M1, canine Madin-Darby canine kidney and human A549 cells. mAb epitopes were mapped by overlapping peptides and selective reactivity to the newly described viral NS3 protein. These mAbs detected NS1 in both the cytoplasm and nucleus by immunostaining, and some detected NS1 as early as 5 h post-infection, suggesting their potential diagnostic use for tracking productive IAV replication and characterizing NS1 structure and function. It was also demonstrated that the newly identified NS3 protein is localized in the cytoplasm to high levels.
authors.affiliation: Manitoba Centre for Proteomics and Systems Biology, Room 799, 715 McDermot Avenue, Winnipeg, MB R3E 3P4, Canada.|Department of Medical Microbiology, Faculty of Medicine, University of Manitoba, Winnipeg, MB R3E 0J6, Canada.
authors.forename: Md Niaz
authors.initials: MN
authors.lastname: Rahim
authors.affiliation: Emerging Pathogens Research Centre, University of Ottawa, Ottawa, ON K1H 8M5, Canada.|Department of Biochemistry, Microbiology, and Immunology, University of Ottawa, Ottawa, ON K1H 8M5, Canada.
authors.forename: Mohammed
authors.initials: M
authors.lastname: Selman
authors.affiliation: Manitoba Centre for Proteomics and Systems Biology, Room 799, 715 McDermot Avenue, Winnipeg, MB R3E 3P4, Canada.
authors.forename: Patricia J
authors.initials: PJ
authors.lastname: Sauder
authors.affiliation: Emerging Pathogens Research Centre, University of Ottawa, Ottawa, ON K1H 8M5, Canada.|Department of Biochemistry, Microbiology, and Immunology, University of Ottawa, Ottawa, ON K1H 8M5, Canada.
authors.forename: Nicole E
authors.initials: NE
authors.lastname: Forbes
authors.affiliation: Emerging Pathogens Research Centre, University of Ottawa, Ottawa, ON K1H 8M5, Canada.|Department of Biochemistry, Microbiology, and Immunology, University of Ottawa, Ottawa, ON K1H 8M5, Canada.
authors.forename: William
authors.initials: W
authors.lastname: Stecho
authors.affiliation: Manitoba Centre for Proteomics and Systems Biology, Room 799, 715 McDermot Avenue, Winnipeg, MB R3E 3P4, Canada.|Department of Medical Microbiology, Faculty of Medicine, University of Manitoba, Winnipeg, MB R3E 0J6, Canada.
authors.forename: Wanhong
authors.initials: W
authors.lastname: Xu
authors.affiliation: Manitoba Centre for Proteomics and Systems Biology, Room 799, 715 McDermot Avenue, Winnipeg, MB R3E 3P4, Canada.|Department of Medical Microbiology, Faculty of Medicine, University of Manitoba, Winnipeg, MB R3E 0J6, Canada.
authors.forename: Mark
authors.initials: M
authors.lastname: Lebar
authors.affiliation: Emerging Pathogens Research Centre, University of Ottawa, Ottawa, ON K1H 8M5, Canada.|Department of Biochemistry, Microbiology, and Immunology, University of Ottawa, Ottawa, ON K1H 8M5, Canada.
authors.forename: Earl G
authors.initials: EG
authors.lastname: Brown
authors.affiliation: Manitoba Institute of Child Health, Room 513, John Buhler Research Centre, 715 McDermot Avenue, Winnipeg, MB R3E 3P4, Canada.|Manitoba Centre for Proteomics and Systems Biology, Room 799, 715 McDermot Avenue, Winnipeg, MB R3E 3P4, Canada.|Department of Medical Microbiology, Faculty of Medicine, University of Manitoba, Winnipeg, MB R3E 0J6, Canada.
authors.forename: Kevin M
authors.initials: KM
authors.lastname: Coombs
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D000914:Antibodies, Viral / Q000276:immunology*; D000918:Antibody Specificity; D002460:Cell Line; D004285:Dogs; D018604:Epitope Mapping; D006801:Humans; D009980:Influenza A virus / Q000145:classification* / Q000276:immunology*; D051379:Mice; D008958:Models, Molecular; D008969:Molecular Sequence Data; D010802:Phylogeny; D011487:Protein Conformation; D017361:Viral Nonstructural Proteins / Q000276:immunology*
chemical_list: D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral; C085820:INS1 protein, influenza virus; D017361:Viral Nonstructural Proteins