tetano
Editor, Senior Moderator
iomolecules. 2015 Jul 16;5(3):1480-1498. [h=1]Characterization of Receptor Binding Profiles of Influenza A Viruses Using An Ellipsometry-Based Label-Free Glycan Microarray Assay Platform.[/h] Fei Y[SUP]1,[/SUP][SUP]2[/SUP], Sun YS[SUP]3,[/SUP][SUP]4[/SUP], Li Y[SUP]5[/SUP], Yu H[SUP]6[/SUP], Lau K[SUP]7[/SUP], Landry JP[SUP]8[/SUP], Luo Z[SUP]9[/SUP], Baumgarth N[SUP]10[/SUP], Chen X[SUP]11[/SUP], Zhu X[SUP]12[/SUP].
[h=3]Author information[/h]
[h=3]Abstract[/h] A key step leading to influenza viral infection is the highly specific binding of a viral spike protein, hemagglutinin (HA), with an extracellular glycan receptor of a host cell. Detailed and timely characterization of virus-receptor binding profiles may be used to evaluate and track the pandemic potential of an influenza virus strain. We demonstrate a label-free glycan microarray assay platform for acquiring influenza virus binding profiles against a wide variety of glycan receptors. By immobilizing biotinylated receptors on a streptavidin-functionalized solid surface, we measured binding curves of five influenza A virus strains with 24 glycans of diverse structures and used the apparent equilibrium dissociation constants (avidity constants, 10-100 pM) as characterizing parameters of viral receptor profiles. Furthermore by measuring binding kinetic constants of solution-phase glycans to immobilized viruses, we confirmed that the glycan-HA affinity constant is in the range of 10 mM and the reaction is enthalpy-driven.
[h=4]KEYWORDS:[/h] binding profile; biosensors; ellipsometry; glycans; high-throughput; influenza A virus; label-free; microarray; reaction kinetics
PMID: 26193329 [PubMed - as supplied by publisher]
[h=3]Author information[/h]
[h=3]Abstract[/h] A key step leading to influenza viral infection is the highly specific binding of a viral spike protein, hemagglutinin (HA), with an extracellular glycan receptor of a host cell. Detailed and timely characterization of virus-receptor binding profiles may be used to evaluate and track the pandemic potential of an influenza virus strain. We demonstrate a label-free glycan microarray assay platform for acquiring influenza virus binding profiles against a wide variety of glycan receptors. By immobilizing biotinylated receptors on a streptavidin-functionalized solid surface, we measured binding curves of five influenza A virus strains with 24 glycans of diverse structures and used the apparent equilibrium dissociation constants (avidity constants, 10-100 pM) as characterizing parameters of viral receptor profiles. Furthermore by measuring binding kinetic constants of solution-phase glycans to immobilized viruses, we confirmed that the glycan-HA affinity constant is in the range of 10 mM and the reaction is enthalpy-driven.
[h=4]KEYWORDS:[/h] binding profile; biosensors; ellipsometry; glycans; high-throughput; influenza A virus; label-free; microarray; reaction kinetics
PMID: 26193329 [PubMed - as supplied by publisher]