title: Modular glycosphere assays for high-throughput functional characterization of influenza viruses.
abstract: BACKGROUND
  The ongoing global efforts to control influenza epidemics and pandemics require high-throughput technologies to detect, quantify, and functionally characterize viral isolates. The 2009 influenza pandemic as well as the recent in-vitro selection of highly transmissible H5N1 variants have only increased existing concerns about emerging influenza strains with significantly enhanced human-to-human transmissibility. High-affinity binding of the virus hemagglutinin to human receptor glycans is a highly sensitive and stringent indicator of host adaptation and virus transmissibility. The surveillance of receptor-binding characteristics can therefore provide a strong additional indicator for the relative hazard imposed by circulating and newly emerging influenza strains.
  
  
  RESULTS
  Streptavidin-coated microspheres were coated with selected biotinylated glycans to mimic either human or avian influenza host-cell receptors. Such glycospheres were used to selectively capture influenza virus of diverse subtypes from a variety of samples. Bound virus was then detected by fluorescently labelled antibodies and analyzed by quantitative flow cytometry. Recombinant hemagglutinin, inactivated virus, and influenza virions were captured and analyzed with regards to receptor specificity over a wide range of analyte concentration. High-throughput analyses of influenza virus produced dose-response curves that allow for functional assessment of relative receptor affinity and thus transmissibility.
  
  
  CONCLUSIONS
  Modular glycosphere assays for high-throughput functional characterization of influenza viruses introduce an important tool to augment the surveillance of clinical and veterinarian influenza isolates with regards to receptor specificity, host adaptation, and virus transmissibility.
authors.affiliation: Singapore-MIT Alliance for Research and Technology, Singapore 138602, Singapore.
authors.forename: Sven N
authors.initials: SN
authors.lastname: Hobbie
authors.forename: Karthik
authors.initials: K
authors.lastname: Viswanathan
authors.forename: Ido
authors.initials: I
authors.lastname: Bachelet
authors.forename: Udayanath
authors.initials: U
authors.lastname: Aich
authors.forename: Zachary
authors.initials: Z
authors.lastname: Shriver
authors.forename: Vidya
authors.initials: V
authors.lastname: Subramanian
authors.forename: Rahul
authors.initials: R
authors.lastname: Raman
authors.forename: Ram
authors.initials: R
authors.lastname: Sasisekharan
mesh_terms: D000818:Animals; D000914:Antibodies, Viral / Q000032:analysis / Q000737:chemistry; D001710:Biotin / Q000737:chemistry; D001717:Birds; D006388:Hemagglutinins / Q000378:metabolism; D057166:High-Throughput Screening Assays / Q000379:methods*; D006801:Humans; D053118:Influenza A Virus, H1N1 Subtype / Q000302:isolation & purification / Q000378:metabolism; D053124:Influenza A Virus, H5N1 Subtype / Q000302:isolation & purification / Q000378:metabolism; D008863:Microspheres; D009975:Orthomyxoviridae / Q000302:isolation & purification* / Q000378:metabolism*; D011134:Polysaccharides / Q000032:analysis / Q000378:metabolism*; D019809:Streptavidin / Q000737:chemistry; D053585:Virus Attachment
chemical_list: D000914:Antibodies, Viral; D006388:Hemagglutinins; D011134:Polysaccharides; D001710:Biotin; D019809:Streptavidin