Reversed phase monolithic analytical columns for the determination of HA1 subunit of influenza virus haemagglutinin
| Autor: | Matjaž Peterka, Boris Pihlar, Lidija Urbas, Miloš Barut, Aleš Štrancar, Boštjan Košir |
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| Rok vydání: | 2011 |
| Předmět: |
Monolithic HPLC column
viruses Blotting Western Orthomyxoviridae Hemagglutinin Glycoproteins Influenza Virus Sensitivity and Specificity Biochemistry Virus Analytical Chemistry Influenza A Virus H1N1 Subtype Western blot medicine Chromatography High Pressure Liquid Gel electrophoresis Detection limit Chromatography Reverse-Phase Chromatography medicine.diagnostic_test biology Chemistry Influenza A Virus H3N2 Subtype Organic Chemistry virus diseases General Medicine Reversed-phase chromatography Hemagglutinin biology.organism_classification Influenza B virus Linear Models Polystyrenes Electrophoresis Polyacrylamide Gel Protein Binding |
| Zdroj: | Journal of Chromatography A. 1218:2432-2437 |
| ISSN: | 0021-9673 |
| Popis: | Monoliths are chromatographic stationary phases, which were specially designed for efficient purification of large biomolecules, like proteins, viruses and DNA. In this work, the small scale monolithic butyl (C4) and styrene-divinyl benzene (SDVB) columns were applied for reversed phase analyses of various degraded influenza viruses. The binding of the HA1 subunit of haemagglutinin to the monolithic columns was confirmed by sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) and the Western blot. The working linear range was determined as 1.60 × 1010 viral particles/mL to at least 1.64 × 1011 viral particles/mL, the limit of detection was found to be 2.56 × 109 virus particles/mL and the limit of quantification was 5.12 × 109 virus particles/mL. The analytical HPLC method developed with the H1N1 virus was also applicable for the analytics of the HA1 subunit of H3N2 influenza virus and the influenza B virus. |
| Databáze: | OpenAIRE |
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