title: Capture and characterization of influenza A virus from primary samples using glycan bead arrays.
abstract: Influenza A viruses (IAVs) utilize sialylated host glycans as ligands for binding and infection. The glycan-binding preference of IAV hemagglutinin (HA) is an important determinant of host specificity. Propagation of IAV in embryonated chicken eggs and cultured mammalian cells yields viruses with amino acid substitutions in the HA that can alter the binding specificity. Therefore, it is important to determine the binding specificity of IAV directly in primary samples since it reflects the actual tropism of virus in nature. We developed a novel platform for analysis of IAV binding specificity in samples that contain very low virus titers. This platform consists of a high-density flexible glycan display on magnetic beads, which promotes multivalent interactions with the viral HA. Glycan-bound virus is detected by quantifying the viral neuraminidase activity via a fluorogenic reporter, 2'-(4-methylumbelliferyl)-α-d-N-acetylneuraminic acid. This method eliminates the need for labeling the virus and significantly enhances the sensitivity of detection.
authors.affiliation: Department of Pathology, Division of Comparative Pathology and Medicine, UC San Diego, La Jolla, CA, USA. Electronic address: micohen@ucsd.edu.
authors.forename: Miriam
authors.initials: M
authors.lastname: Cohen
authors.affiliation: Department of Chemistry and Biochemistry, UC San Diego, La Jolla, CA, USA.
authors.forename: Christopher J
authors.initials: CJ
authors.lastname: Fisher
authors.affiliation: Department of Chemistry and Biochemistry, UC San Diego, La Jolla, CA, USA.
authors.forename: Mia L
authors.initials: ML
authors.lastname: Huang
authors.affiliation: Department of Pathology, Microbiology and Immunology, School of Veterinary Medicine, UC Davis, Davis, CA, USA.
authors.forename: LeAnn L
authors.initials: LL
authors.lastname: Lindsay
authors.affiliation: Department of Pathology, Microbiology and Immunology, School of Veterinary Medicine, UC Davis, Davis, CA, USA.
authors.forename: Magdalena
authors.initials: M
authors.lastname: Plancarte
authors.affiliation: Department of Pathology, Microbiology and Immunology, School of Veterinary Medicine, UC Davis, Davis, CA, USA.
authors.forename: Walter M
authors.initials: WM
authors.lastname: Boyce
authors.affiliation: Department of Chemistry and Biochemistry, UC San Diego, La Jolla, CA, USA.
authors.forename: Kamil
authors.initials: K
authors.lastname: Godula
authors.affiliation: Department of Pathology, Division of Comparative Pathology and Medicine, UC San Diego, La Jolla, CA, USA. Electronic address: pgagneux@ucsd.edu.
authors.forename: Pascal
authors.initials: P
authors.lastname: Gagneux
mesh_terms: D000818:Animals; D002988:Cloaca / Q000821:virology*; D004285:Dogs; D004372:Ducks / Q000821:virology; D009980:Influenza A virus / Q000302:isolation & purification* / Q000378:metabolism; D061985:Madin Darby Canine Kidney Cells; D008280:Magnetics; D008863:Microspheres; D009077:Mucins; D009439:Neuraminidase / Q000378:metabolism; D011134:Polysaccharides / Q000378:metabolism*; D012680:Sensitivity and Specificity; D014764:Viral Proteins / Q000378:metabolism
chemical_list: D009077:Mucins; D011134:Polysaccharides; D014764:Viral Proteins; C487630:NA protein, influenza A virus; D009439:Neuraminidase
keywords: Glycan array; Glycans; Influenza A; Magnetic beads; Neuraminidase; Sialic acids