YUE Wei, YE Lin-bai, ZHANG Jie and LIU Jing. Preparation of Rabies Vaccine by Perfusion Culture of Vero Cells on Microcarriers[J]. Virologica Sinica, 2004, 19(4): 373-375.
Citation: YUE Wei, YE Lin-bai, ZHANG Jie, LIU Jing. Preparation of Rabies Vaccine by Perfusion Culture of Vero Cells on Microcarriers .VIROLOGICA SINICA, 2004, 19(4) : 373-375.

微载体灌注培养制备Vero细胞狂犬病疫苗

  • 本文研究在生物反应器中用微载体连续灌注培养Vero细胞生产狂犬病毒制备技术。在5L体积的生物反应器中,加入含10g/L微载体的199培养基,接种Vero细胞至细胞浓度达到1×105/mL,培养7d后细胞可生长至6~7×106/mL,然后以感染复数(MOI)为0.01接种狂犬病毒VaG株,接毒后24h开始收获,连续收获12d左右,收获的病毒滴度范围在6.0~8.5log LD50/mL,收获的病毒原液经浓缩、灭活和纯化等步骤制备成疫苗,各项质量指标均达到《中国生物制品规程》2000年版要求。实验表明,用生物反应器微载体灌注培养制备人用Vero细胞狂犬病疫苗小试工艺可行。

Preparation of Rabies Vaccine by Perfusion Culture of Vero Cells on Microcarriers

  • Corresponding author: YE Lin-bai, 
  • To establish a technique for preparing rabies vaccine by perfusion culture on microcarriers in bioreactor, a 5 litre bioreactor was filled with 199 cell culture medium which contains 10g/L of microcarriers. Then Vero cells were inoculated with a final cell density of 1×105/mL . After 7 days of culture, cell density can reach 6~7×106/mL, then rabies virus VaG strain was inoculated with a MOI of 0.01. After 24h inoculation, the cultured viruses were harvested continuously for 12 days. The titer of collected virus in the culture medium was about 6.0 to 8.5 logLD50/mL. From these collected viruses, vaccines were prepared through the steps of concentration, inactivation and purification, The quality of these vaccines meets “The Chinese Requirements for Biologics Year 2000”. This lab-scale method for preparing rabies vaccine by perfusion culture on microcarries is viable.

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    Preparation of Rabies Vaccine by Perfusion Culture of Vero Cells on Microcarriers

      Corresponding author: YE Lin-bai,
    • 1. 1. Institute of virology,Wuhan University,Wuhan 430072,China
    • 2. Wuhan Institute of Biological Products,Wuhan 430060,China

    Abstract: To establish a technique for preparing rabies vaccine by perfusion culture on microcarriers in bioreactor, a 5 litre bioreactor was filled with 199 cell culture medium which contains 10g/L of microcarriers. Then Vero cells were inoculated with a final cell density of 1×105/mL . After 7 days of culture, cell density can reach 6~7×106/mL, then rabies virus VaG strain was inoculated with a MOI of 0.01. After 24h inoculation, the cultured viruses were harvested continuously for 12 days. The titer of collected virus in the culture medium was about 6.0 to 8.5 logLD50/mL. From these collected viruses, vaccines were prepared through the steps of concentration, inactivation and purification, The quality of these vaccines meets “The Chinese Requirements for Biologics Year 2000”. This lab-scale method for preparing rabies vaccine by perfusion culture on microcarries is viable.

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