Purification of Vero cell derived live replication deficient influenza A and B virus by ion exchange monolith chromatography

We explored the possibilities for purification of various Delta NS1 live, replication deficient influenza viruses on ion exchange methacrylate monoliths. Influenza A Delta NSI-H1N1, Delta NSI-H3N2, Delta NS1-H5N1 and Delta NS-Iinfluenza B viruses were propagated in Vero cells and concentrated by tangential flow filtration. All four virus strains adsorbed well to CIM QA and CIM DEAE anion exchangers, with CIM QA producing higher recoveries than CIM DEAE. Delta NSI-influenza A viruses adsorbed well also to CIM SO3 cation exchanger at the same pH, while Delta NSI-influenza B virus adsorption to CIM SO3 was not complete. Dynamic binding capacity (DBC) for CIM QA, DEAE and SO3 methacrylate monoliths for influenza A Delta NSI-H1N1 virus were 1.9E + 10 TCID50/ml, 1.0E + 10 TCID50/ml and 8.9E + 08 TCID50/ml, respectively. Purification of Delta NSI viruses on CIM QA was scaled up and reproducibility was confirmed. Yields of infectious virus on CIM QA were between 70.8 +/- 32.3% and 87 +/- 30.8%. Total protein removal varied from 93.3 +/- 0.4% to 98.6 +/- 0.2% and host cell DNA removal efficiency was ranging from 76.4% to 99.9% and strongly depended on pretreatment with deoxyribonuclease. (C) 2014 Elsevier Ltd. All rights reserved.