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KMID : 0545120000100050728
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Journal of Microbiology and Biotechnology
2000 Volume.10 No. 5 p.728 ~ p.732
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Optimization of Propagation of Anagrapha falcifera Nuclear Polyhedrosis Virus in Spodoptera frugiperda 21 Cells
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Lee, Jong Min
Chang, Kyung Hwa/Park, Jin O/Park, Jong Hwa/Hwang, In Sook/Lee, Youn Hyung/Yang, Jai Myung
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Abstract
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Propagation of Anagrapha falcifera nuclear polyhedrosis virus (AfNPV) was investigated using well-plates and split-flow air-lift bioreactors. In well-plate experiments, the effects of pH, cell density at a point of infection, serum concentration, DEAE-dextran, and lipid on virus propagation were all closely examined, The AfNPV titer in well-plates was optimal at pH 6.8 and 3×l0 exp (6) cells/㎠. The virus titer was not dramatically affected when the fetal bovine serum concentration was reduced from 10% to 5%. The addition of cholesterol at AfNPV infection of Sf21 cells enhanced the virus titer, whereas the addition of DEAE-dextran did not improve the titer. The AfNPV titer (3.8×10 exp (7) TCID_50/㎖) at optimized conditions for well-plate experiments was 2.5-fold higher than for the control. In bioreactor experiments, the AfNPV titer showed its maximum level at air flow rates of 20-40㎖/min. In a split-flow air-lift bioreactor, AfNPV titer (2.3×10 exp (7) TCID50/㎖) was 1.5-fold higher than the control when the culture was at pH 6.8 and supplemented with 0.34mM cholesterol.
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