Quantification of Receptor Association, Dissociation, and NA-Dependent Motility of Influenza A Particles by Biolayer Interferometry

Erik de Vries, Hongbo Guo, Wenjuan Du, Mengying Liu, Frank J M van Kuppeveld, Cornelis A M de Haan

Research output: Chapter in Book/Report/Conference proceedingChapterAcademicpeer-review

Abstract

We describe a method for real-time analysis and quantification of influenza A virus (IAV)-receptor interactions by biolayer interferometry (BLI). Biotinylated synthetic sialoglycans or sialoglycoproteins (biotinylated or Fc-tagged) were immobilized on the tip of biosensors (coated with streptavidin or protein A) that were subsequently dipped into IAV particle solutions in 96-well plates. Association and/or dissociation of IAV particles was recorded in consecutive steps in buffers of choice. From the association and dissociation curves, parameters can be derived that describe IAV particle-receptor interactions in absence or presence of neuraminidase activity. Overall, the method provides a quantitative description of the hemagglutinin-neuraminidase balance that determines the interaction kinetics of IAV with specific sialoglycan receptors.

Original languageEnglish
Title of host publicationGlycovirology
Subtitle of host publicationMethods and Protocols
PublisherHumana Press
Chapter10
Pages123-140
Number of pages18
ISBN (Electronic)978-1-0716-2635-1
ISBN (Print)978-1-0716-2634-4
DOIs
Publication statusPublished - 30 Sept 2022

Publication series

NameMethods in Molecular Biology
PublisherHumana Press
Volume2556
ISSN (Print)1064-3745

Bibliographical note

Publisher Copyright:
© 2022, The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.

Keywords

  • Neuraminidase
  • Binding kinetics
  • Hemagglutinin
  • Biolayer interferometry
  • Virus motility
  • Association
  • Influenza virus
  • Sialoglycoprotein
  • Sialoglycan
  • Dissociation

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