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Rhinoviruses pp 101-128 | Cite as

Capillary Electrophoresis, Gas-Phase Electrophoretic Mobility Molecular Analysis, and Electron Microscopy: Effective Tools for Quality Assessment and Basic Rhinovirus Research

  • Victor U. Weiss
  • Xavier Subirats
  • Mohit Kumar
  • Shushan Harutyunyan
  • Irene Gösler
  • Heinrich Kowalski
  • Dieter BlaasEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1221)

Abstract

We describe standard methods for propagation, purification, quality control, and physicochemical characterization of human rhinoviruses, using HRV-A2 as an example. Virus is propagated in HeLa-OHIO cells grown in suspension culture and purified via sucrose density gradient centrifugation. Purity and homogeneity of the preparations are assessed with SDS-polyacrylamide gel electrophoresis (SDS-PAGE), capillary electrophoresis (CE), gas-phase electrophoretic mobility molecular analysis (GEMMA), and electron microscopy (EM). We also briefly describe usage of these methods for the characterization of subviral particles as well as for the analysis of their complexes with antibodies and soluble recombinant receptor mimics.

Key words

Rhinovirus Purification Analysis Capillary electrophoresis Gas-phase electrophoretic mobility molecular analysis Electron microscopy 

Notes

Acknowledgements

The work described in this chapter was made possible via various grants and the DK “Structure and Interaction of Macromolecules” funded by the Austrian Science Fund (FWF). We thank Guenter Resch (Campus Science Support Facilities) for help with the electron microscope and Günter Allmaier (Techn. Univ. Vienna) for making available the GEMMA instrument.

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Copyright information

© Springer Science+Business Media New York 2015

Authors and Affiliations

  • Victor U. Weiss
    • 1
  • Xavier Subirats
    • 2
  • Mohit Kumar
    • 3
  • Shushan Harutyunyan
    • 3
  • Irene Gösler
    • 3
  • Heinrich Kowalski
    • 3
  • Dieter Blaas
    • 3
    Email author
  1. 1.Institute of Chemical Technologies and AnalyticsVienna University of TechnologyViennaAustria
  2. 2.Department of Analytical ChemistryUniversity of BarcelonaBarcelonaSpain
  3. 3.Max F. Perutz Laboratories, Department of Medical BiochemistryMedical University of ViennaViennaAustria

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