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| Bioprocess Optimization for Recombinant Herpes Simplex Virus type Ⅱ Vaccine Production Based Cell Microcarrier Suspention Culture |
| LIN Mei-ling, TANG Yin, ZHANG Ming, YI Xiao-ping, HUANG Ming-zhi |
| The State Laboratory Bioreactor Engineering, East China University of Science and Technology, Shanghai 200237, China |
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Abstract The attenuated recombinant herpes simplex virus type Ⅱ(rHSV-Ⅱ) have a good oncolytic effect in vitro, and showed higher efficacy against B16R melanoma in mice. As an efficient new oncolytic virus, rHSV-Ⅱ is expected for the clinical treatment of cancer. African Green Monkey Kidney (Vero) Cells have been widely used for human vaccine production since they were sensitive and high production hosts, and safety in micro-carrier culture. With the self-development chemically defined serum-free medium added 1% NBCS, the bead-to-dead transfer of vero cells in bioreactor was developed. The optimal condition of transfer were about 30 cells/MC inoculated, cultured for 60 h when the cells density reached about the same density with the initial inoculated, the static time/stiring time was 42 min/3min. It can be achieved at least four times successive transfer for bioreactor amplification by the 1:4 (V:V), cells density reach up to 6.0×106 cells/ml. Under these conditions, the bioreactor culture process of rHSV-Ⅱ in microcarrier suspension culture has been developed, maximum virus titer was reached up to 6.62 lgTCID50/ml. A simple, efficient process method is provided for large-scale production of anti-tumor vaccine of rHSV-Ⅱ.
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Received: 26 December 2013
Published: 25 March 2014
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