Capillary electrophoresis, gas-phase electrophoretic mobility molecular analysis, and electron microscopy: effective tools for quality assessment and basic rhinovirus research

Methods Mol Biol. 2015:1221:101-28. doi: 10.1007/978-1-4939-1571-2_9.

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Biomedical Research / methods
  • Cell Culture Techniques
  • Centrifugation, Density Gradient
  • Electrophoresis, Capillary / methods*
  • Electrophoresis, Polyacrylamide Gel
  • Electrophoretic Mobility Shift Assay / methods
  • HeLa Cells / virology
  • Humans
  • Microscopy, Electron / methods*
  • Microscopy, Electron, Transmission / methods
  • Rhinovirus* / growth & development
  • Rhinovirus* / isolation & purification