tetano
Editor, Senior Moderator
Biotechnol Prog. 2012 Jul 27. doi: 10.1002/btpr.1601. [Epub ahead of print]
Real-time monitoring of influenza virus production kinetics in HEK293 cell cultures.
Emma P, Kamen A.
Source
6100 Royalmount Ave, Montreal, Quebec, Canada, H4P 2R2.
Abstract
There is an increased interest from the vaccine Industry to use mammalian cell cultures for influenza vaccine manufacturing. Therefore, it became important to study the influenza infection mechanism, the viral-host interaction and the replication kinetics from a bioprocessing stand point to maximize the influenza viral production yield in cell culture. In the present study, influenza replication kinetics in HEK293 cells was studied. Two infection conditions were evaluated, a low (0.01) and a high multiplicity of infection (1.0). Critical time points of the viral production cycle (infection, protein synthesis, viral assembly and budding, viral release, host-cell death) were identified in small-scale cell cultures. Additionally, cell growth, viability, viral titers were monitored in the viral production process. The infection state of the cultivated cell population was assessed by influenza immunolabeling throughout the culture period. Influenza virus production kinetics were also on-line monitored by dielectric spectroscopy and successfully correlated to real-time capacitance measures. Overall, this work provided insights into the mechanisms associated with the infection of human HEK293 cell lines by the influenza virus and demonstrated, once again, the usefulness of multi-frequency scanning permittivity for in-line monitoring and supervision of cell-based viral production processes. ? 2012 American Institute of Chemical Engineers Biotechnol. Prog., 2012.
Copyright ? 2012 American Institute of Chemical Engineers (AIChE).
PMID:
22848016
[PubMed - as supplied by publisher]
http://www.ncbi.nlm.nih.gov/pubmed/22848016
Real-time monitoring of influenza virus production kinetics in HEK293 cell cultures.
Emma P, Kamen A.
Source
6100 Royalmount Ave, Montreal, Quebec, Canada, H4P 2R2.
Abstract
There is an increased interest from the vaccine Industry to use mammalian cell cultures for influenza vaccine manufacturing. Therefore, it became important to study the influenza infection mechanism, the viral-host interaction and the replication kinetics from a bioprocessing stand point to maximize the influenza viral production yield in cell culture. In the present study, influenza replication kinetics in HEK293 cells was studied. Two infection conditions were evaluated, a low (0.01) and a high multiplicity of infection (1.0). Critical time points of the viral production cycle (infection, protein synthesis, viral assembly and budding, viral release, host-cell death) were identified in small-scale cell cultures. Additionally, cell growth, viability, viral titers were monitored in the viral production process. The infection state of the cultivated cell population was assessed by influenza immunolabeling throughout the culture period. Influenza virus production kinetics were also on-line monitored by dielectric spectroscopy and successfully correlated to real-time capacitance measures. Overall, this work provided insights into the mechanisms associated with the infection of human HEK293 cell lines by the influenza virus and demonstrated, once again, the usefulness of multi-frequency scanning permittivity for in-line monitoring and supervision of cell-based viral production processes. ? 2012 American Institute of Chemical Engineers Biotechnol. Prog., 2012.
Copyright ? 2012 American Institute of Chemical Engineers (AIChE).
PMID:
22848016
[PubMed - as supplied by publisher]
http://www.ncbi.nlm.nih.gov/pubmed/22848016