Monolithic anion-exchange chromatography yields rhinovirus of high purity
| Autor: | Christina Bliem, Dieter Blaas, Günter Allmaier, Heinrich Kowalski, Victor U. Weiss, Irene Gösler, Thomas Dechat, Sofiya Fedosyuk |
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| Rok vydání: | 2018 |
| Předmět: |
0301 basic medicine
Electrospray Chromatography Rhinovirus Picornavirus biology 010401 analytical chemistry Chromatography Ion Exchange biology.organism_classification 01 natural sciences Negative stain Article 0104 chemical sciences 03 medical and health sciences chemistry.chemical_compound Electrophoresis 030104 developmental biology Capillary electrophoresis chemistry Peptide mass fingerprinting Virology Sodium dodecyl sulfate Polyacrylamide gel electrophoresis |
| Zdroj: | Journal of Virological Methods. 251:15-21 |
| ISSN: | 0166-0934 |
| Popis: | For vaccine development, 3D-structure determination, direct fluorescent labelling, and numerous other studies, homogeneous virus preparations of high purity are essential. Working with human rhinoviruses (RVs), members of the picornavirus family and the main cause of generally mild respiratory infections, we noticed that our routine preparations appeared highly pure on analysis by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), exclusively showing the four viral capsid proteins (VPs). However, the preparations turned out to contain substantial amounts of contaminating material when analyzed by orthogonal analytical methods including capillary zone electrophoresis, nano electrospray gas-phase electrophoretic mobility molecular analysis (nES GEMMA), and negative stain transmission electron microscopy (TEM). Because these latter analyses are not routine to many laboratories, the above contaminations might remain unnoticed and skew experimental results. By using human rhinovirus serotype A2 (RV-A2) as example we report monolithic anion-exchange chromatography (AEX) as a last polishing step in the purification and demonstrate that it yields infective, highly pure, virus (RV-A2 in the respective fractions was confirmed by peptide mass fingerprinting) devoid of foreign material as judged by the above criteria. |
| Databáze: | OpenAIRE |
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