title: Identification of human single-chain antibodies with broad reactivity for noroviruses.
abstract: Norovirus infections are a common cause of gastroenteritis and new methods to rapidly diagnose norovirus infections are needed. The goal of this study was to identify antibodies that have broad reactivity of binding to various genogroups of norovirus. A human scFv phage display library was used to identify two antibodies, HJT-R3-A9 and HJT-R3-F7, which bind to both genogroups I and II norovirus virus-like particles (VLPs). Mapping experiments indicated that the HJT-R3-A9 clone binds to the S-domain while the HJT-R3-F7 clone binds the P-domain of the VP1 capsid protein. In addition, a family of scFv antibodies was identified by elution of phage libraries from the GII.4 VLP target using a carbohydrate that serves as an attachment factor for norovirus on human cells. These antibodies were also found to recognize both GI and GII VLPs in enzyme-linked immunosorbent assay (ELISA) experiments. The HJT-R3-A9, HJT-R3-F7 and scFv antibodies identified with carbohydrate elution were shown to detect antigen from a clinical sample known to contain GII.4 norovirus but not a negative control sample. Finally, phages displaying the HJT-R3-A9 scFv can be used directly to detect both GI.1 and GII.4 norovirus from stool samples, which has the potential to simplify and reduce the cost of diagnostics based on antibody-based ELISA methods.
authors.affiliation: Department of Pharmacology, Baylor College of Medicine, Houston, TX, USA.
authors.forename: Wanzhi
authors.initials: W
authors.lastname: Huang
authors.affiliation: Department of Pharmacology, Baylor College of Medicine, Houston, TX, USA.
authors.forename: Moumita
authors.initials: M
authors.lastname: Samanta
authors.affiliation: Department of Molecular Virology and Microbiology, Baylor College of Medicine, Houston, TX, USA.
authors.forename: Sue E
authors.initials: SE
authors.lastname: Crawford
authors.affiliation: Department of Molecular Virology and Microbiology, Baylor College of Medicine, Houston, TX, USA Department of Medicine, Baylor College of Medicine, Houston, TX, 77030, USA.
authors.forename: Mary K
authors.initials: MK
authors.lastname: Estes
authors.affiliation: Department of Molecular Virology and Microbiology, Baylor College of Medicine, Houston, TX, USA.
authors.forename: Frederick H
authors.initials: FH
authors.lastname: Neill
authors.affiliation: Department of Molecular Virology and Microbiology, Baylor College of Medicine, Houston, TX, USA Department of Medicine, Baylor College of Medicine, Houston, TX, 77030, USA.
authors.forename: Robert L
authors.initials: RL
authors.lastname: Atmar
authors.affiliation: Department of Pharmacology, Baylor College of Medicine, Houston, TX, USA Department of Molecular Virology and Microbiology, Baylor College of Medicine, Houston, TX, USA timothyp@bcm.edu.
authors.forename: Timothy
authors.initials: T
authors.lastname: Palzkill
mesh_terms: D000595:Amino Acid Sequence; D000914:Antibodies, Viral / Q000737:chemistry / Q000276:immunology* / Q000378:metabolism; D001666:Binding Sites, Antibody; D004926:Escherichia coli; D006801:Humans; D008969:Molecular Sequence Data; D029322:Norovirus / Q000276:immunology* / Q000302:isolation & purification*; D019151:Peptide Library; D011485:Protein Binding; D057127:Single-Chain Antibodies / Q000737:chemistry / Q000276:immunology* / Q000378:metabolism; D014771:Virion / Q000276:immunology / Q000378:metabolism
chemical_list: D000914:Antibodies, Viral; D019151:Peptide Library; D057127:Single-Chain Antibodies
keywords: antibody library; diagnostics; norovirus; phage display; single-chain antibody