Development of a Strategy of Influenza Virus Separation Based on Pseudoaffinity Chromatography on Short Monolithic Columns
- 14 February 2008
- journal article
- Published by American Chemical Society (ACS) in Analytical Chemistry
- Vol. 80 (6), 2188-2198
- https://doi.org/10.1021/ac702258t
Abstract
This research is devoted to the development and optimization of fine purification processes realized on short monolithic columns (CIM disks), using influenza vaccine and viruslike synthetic particles as model objects. The pseudoaffinity mode of liquid chromatography has been used as a tool for dynamic adsorption experiments. Viruslike particles, close to the dimensions of influenza viruses, were developed by means of main antigen of influenza viruses (hemeagglutinin) covalent binding to the outer aminated surface of synthetic latex particles. The natural receptor analogues of sialic acid were used as affinity ligands immobilized on the surface of the CIM disk by different ways to achieve a high adsorption capacity. Also, some other ligands were tested as possible candidates for virus capturing. The affinity binding parameters for influenza A virus were obtained by frontal elution method at optimized chromatographic conditions and immobilization schemes. The experimental data pointed out the possibility of selective isolation of hemeagglutinin from a mixture of vaccine proteins. The results obtained by fast affinity chromatography have shown functional and sterical correspondence viruslike synthetic models to influenza viruses. Additionally, the optimization of chromatographic conditions allowed isolation of influenza virus A while maintaining its virulence. The maximum value of adsorption capacity was registered for a monolithic disk, modified subsequently by chitosan and 2,6-sialyllactose and found to be equal to 6.9 × 1012 virions/mL support.Keywords
This publication has 26 references indexed in Scilit:
- Study of dynamic adsorption behavior of large-size protein-bearing particlesJournal of Chromatography A, 2007
- Lectin-affinity chromatography for downstream processing of MDCK cell culture derived human influenza A virusesVaccine, 2007
- Purification of cell culture‐derived human influenza A virus by size‐exclusion and anion‐exchange chromatographyBiotechnology & Bioengineering, 2006
- Convective Interaction Media short monolithic columns: Enabling chromatographic supports for the separation and purification of large biomoleculesJournal of Separation Science, 2005
- Chromatographic media for bioseparationJournal of Chromatography A, 2005
- Screening of inhibitors for influenza A virus using high-performance affinity chromatography and combinatorial peptide librariesJournal of Chromatography A, 2005
- Concentration of plant viruses using monolithic chromatographic supportsJournal of Virological Methods, 2004
- Application of short monolithic columns for improved detection of virusesJournal of Virological Methods, 2003
- Very fast analysis of impurities in immunoglobulin concentrates using conjoint liquid chromatography on short monolithic disksJournal of Immunological Methods, 2002
- Application of monoliths as supports for affinity chromatography and fast enzymatic conversionJournal of Biochemical and Biophysical Methods, 2001