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Purification of mumps and measles virus by hydrophobic interaction chromatography on monolithic columns

Mumps (MuV) and measles viruses (MeV) are nonsegmented, negative stranded RNA viruses of the family Paramyxoviridae, subfamily Paramyxovirinae. MuV and MeV are the cause of mumps and measles disease, both preventable by vaccination. Purity of virus particles used for vaccines or gene therapy is of utmost importance in order to obtain a product with high purity that can meet the stringent guidelines of the regulatory authorities for potency and safety. Chromatography is being increasingly used in virus purification, especially since the development of new chromatographic matrices. Here, research on MuV and MeV purification by hydrophobic interaction chromatography (HIC) on monolithic column is presented. The influence of different ammonium sulfate concentrations in the binding buffer on purification efficacy as well as column channel size was examined. Recovery of viable viruses was determined by 50% cell culture infective dose assay. Size and concentration of total virus particles were determined by Nanoparticle Tracking Analysis using NanoSight instrument and elution fractions were tested for Vero cell proteins by ELISA. Results show that total particle concentration, as well as the concentration of live viruses increases with decreasing ammonium sulfate concentration in the elution buffer. High ammonium sulfate concentrations did not induce virus aggregation but did induce some loss of viable particles. Decrease of MuV and MeV titers was also observed upon loading the virus on chromatographic system without the column. Viable virus recovery was moderately high (up to 40 %), therefore HIC appears to be a promising method for purification of viable MeV and MuV.

Mumps virus ; Measles virus ; Hydrophobic interaction chromatography ; Monoliths ; Nanoparticle Tracking Analysis

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