title: Characterization of a broadly reactive monoclonal antibody against norovirus genogroups I and II: recognition of a novel conformational epitope.
abstract: Norovirus, which belongs to the family Caliciviridae, is one of the major causes of nonbacterial acute gastroenteritis in the world. The main human noroviruses are of genogroup I (GI) and genogroup II (GII), which were subdivided further into at least 15 and 18 genotypes (GI/1 to GI/15 and GII/1 to GII/18), respectively. The development of immunological diagnosis for norovirus had been hindered by the antigen specificity of the polyclonal antibody. Therefore, several laboratories have produced broadly reactive monoclonal antibodies, which recognize the linear GI and GII cross-reactive epitopes or the conformational GI-specific epitope. In this study, we characterized the novel monoclonal antibody 14-1 (MAb14-1) for further development of the rapid immunochromatography test. Our results demonstrated that MAb14-1 could recognize 15 recombinant virus-like particles (GI/1, 4, 8, and 11 and GII/1 to 7 and 12 to 15) and showed weak affinity to the virus-like particle of GI/3. This recognition range is the broadest of the existing monoclonal antibodies. The epitope for MAb14-1 was identified by fragment, sequence, structural, and mutational analyses. Both terminal antigenic regions (amino acid positions 418 to 426 and 526 to 534) on the C-terminal P1 domain formed the conformational epitope and were in the proximity of the insertion region (positions 427 to 525). These regions contained six amino acids responsible for antigenicity that were conserved among genogroup(s), genus, and Caliciviridae. This epitope mapping explained the broad reactivity and different titers among GI and GII. To our knowledge, we are the first group to identify the GI and GII cross-reactive monoclonal antibody, which recognizes the novel conformational epitope. From these data, MAb14-1 could be used further to develop immunochromatography.
authors.affiliation: Department of Developmental Medical Sciences, Graduate School of Medicine, The University of Tokyo, 7-3-1 Hongo, Tokyo, Japan.
authors.forename: Tomoyuki
authors.initials: T
authors.lastname: Shiota
authors.forename: Michio
authors.initials: M
authors.lastname: Okame
authors.forename: Sayaka
authors.initials: S
authors.lastname: Takanashi
authors.forename: Pattara
authors.initials: P
authors.lastname: Khamrin
authors.forename: Makiko
authors.initials: M
authors.lastname: Takagi
authors.forename: Kenji
authors.initials: K
authors.lastname: Satou
authors.forename: Yuichi
authors.initials: Y
authors.lastname: Masuoka
authors.forename: Fumihiro
authors.initials: F
authors.lastname: Yagyu
authors.forename: Yuko
authors.initials: Y
authors.lastname: Shimizu
authors.forename: Hideki
authors.initials: H
authors.lastname: Kohno
authors.forename: Masashi
authors.initials: M
authors.lastname: Mizuguchi
authors.forename: Shoko
authors.initials: S
authors.lastname: Okitsu
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Ushijima
mesh_terms: D000595:Amino Acid Sequence; D000911:Antibodies, Monoclonal / Q000276:immunology*; D000914:Antibodies, Viral / Q000276:immunology*; D002213:Capsid / Q000737:chemistry*; D003429:Cross Reactions; D018604:Epitope Mapping; D000939:Epitopes / Q000737:chemistry* / Q000235:genetics / Q000276:immunology; D006801:Humans; D008969:Molecular Sequence Data; D029322:Norovirus / Q000276:immunology / Q000302:isolation & purification*; D017354:Point Mutation; D011487:Protein Conformation
chemical_list: D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral; D000939:Epitopes