Artículos de revistas
Statistical optimization of influenza H1N1 production from batch cultures of suspension Vero cells (sVero)
Fecha
2011-07Registro en:
Paillet, Cristian; Forno, Guillermina; Soldano, Nicolas; Kratje, Ricardo Bertoldo; Etcheverrigaray, Marina; Statistical optimization of influenza H1N1 production from batch cultures of suspension Vero cells (sVero); Elsevier; Vaccine; 29; 41; 7-2011; 7212-7217
0264-410X
Autor
Paillet, Cristian
Forno, Guillermina
Soldano, Nicolas
Kratje, Ricardo Bertoldo
Etcheverrigaray, Marina
Resumen
Efficient vaccine production requires the growth of large quantities of virus produced with high yield from a safe host system. Human influenza vaccines are produced in embryonated chicken eggs. However, over the last decade many efforts have allowed the establishment of cell culture-derived vaccines.
We generated a Vero cell line adapted to grow in suspension (sVero) in a serum-free medium and evaluated it for its potential as host cell for influenza vaccine production. Initially we studied the capacity of sVero cells to grow in the presence of incremental concentrations of trypsin. In comparison with adherent Vero cells (aVero), we found that sVero cells maintain their growth kinetics even with a threefold increase in trypsin concentration. The influence of the conditions of infection on the yield of H1N1 produced in serum-free suspension cultures of sVero cells was investigated by a 22 full factorial experiment with center point. Each experiment tested the influence of the multiplicity of infection (m.o.i.) and trypsin concentration, on production yields at two levels, in four possible combinations of levels and conditions, plus a further combination in which each condition was set in the middle of its extreme levels. On the basis of software analysis, a combination of m.o.i. of 0.0066 TCID50%/cell and trypsin concentration of 5 g/1.0 × 106 cells with a desirability of 0.737 was selected as the optimized condition for H1N1
production in sVero cells. Our results show the importance of proper selection of infection conditions for H1N1 production on sVero cells in serum-free medium.