Impact of adsorbents selection on capture efficiency of cell culture derived human influenza viruses.
The study aims on affinity matrix selection for a cell culture derived influenza virus capture step in downstream processing. Euonymus europaeus lectin (EEL) was used as an affinity ligand. Human influenza A/Puerto Rico/8/34 (H1N1) virus produced in MDCK cells was chosen as a model strain. The chromatographic separation characteristics of reinforced cellulose membranes and different matrices such as agarose, cellulose, polymer and glass particles with immobilized EEL have been determined. Results obtained were compared to affinity matrices, which are currently used in large-scale vaccine manufacturing. Mass balances for the viral membrane protein hemagglutinin showed that EEL affinity chromatography results in higher recoveries than conventional processes using Cellufine sulphate and heparinized agarose. The most efficient media, a polymer and a cellulose membrane, have been further characterized by protein and host cell DNA measurements. Separations based on the polymer matrix and the cellulose membrane removed contaminating DNA to 0.2 and 1%, respectively. Total protein contents were decreased to 50 and 31%, respectively. The EEL-membrane showed the highest influenza virus binding capacity. These characteristics demonstrate that EEL affinity chromatography is a promising candidate for capturing influenza viruses from MDCK cell culture broths in addition to currently applied chromatographic media.
Opitz
Lars
L
Bioprocess Engineering, Max Planck Institute for Dynamics of Complex Technical Systems, Sandtorstrasse 1, 39106 Magdeburg, Germany. opitz@mpi-magdeburg.mpg.de
Lehmann
Sylvia
S
Zimmermann
Anke
A
Reichl
Udo
U
Wolff
Michael W
MW
D000042:Absorption; D001672:Biocompatible Materials / Q000737:chemistry*; D002846:Chromatography, Affinity / Q000379:methods*; D006801:Humans; D053118:Influenza A Virus, H1N1 Subtype / Q000302:isolation & purification* / Q000378:metabolism*; D007251:Influenza, Human / Q000821:virology*; D037102:Lectins / Q000378:metabolism*; D008422:Materials Testing; D014462:Ultrafiltration / Q000379:methods; D014776:Virus Cultivation / Q000379:methods*
D001672:Biocompatible Materials; D037102:Lectins