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 language | English |
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Title of host publication | Glycovirology |
Subtitle of host publication | Methods and Protocols |
Publisher | Humana Press |
Chapter | 10 |
Pages | 123-140 |
Number of pages | 18 |
ISBN (Electronic) | 978-1-0716-2635-1 |
ISBN (Print) | 978-1-0716-2634-4 |
DOIs | |
Publication status | Published - 30 Sept 2022 |
Publication series
Name | Methods in Molecular Biology |
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Publisher | Humana Press |
Volume | 2556 |
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