PK-15细胞微载体悬浮培养生产猪细小病毒的工艺研究

doi:10.13523/j.cb.20150709

中国生物工程杂志

2015, Vol. 35

Issue (7): 62-67 DOI: 10.13523/j.cb.20150709

技术与方法

PK-15细胞微载体悬浮培养生产猪细小病毒的工艺研究

李智力, 易小萍, 储炬, 庄英萍, 张嗣良

华东理工大学生物反应器工程国家重点实验室上海 200237

A Microcarrier Cell Culture Process for propagating Porcine Parvovirus in PK-15 Cells

LI Zhi-li, YI Xiao-ping, CHU Ju, ZHUANG Ying-ping, ZHANG Si-liang

East China University of Science and Technology, State Key Laboratory of Bioreactor Engineering, Shanghai 200237, China

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摘要：

通过对猪细小病毒接毒时间TOI、MOI和收毒时间进行优化,开发了一种基于PK-15细胞静置培养的猪细小病毒生产工艺,最大病毒滴度达到10^7.5 TCID₅₀/ml。通过进一步优化接毒时间,成功建立了基于PK-15细胞反应器微载体悬浮培养的猪细小病毒培养工艺,在5L反应器上最大病毒滴度达到10^7.2 TCID₅₀/ml。首次发现乳酸对葡萄糖得率与病毒滴度的正相关性,当猪细小病毒滴度处于最大值时,乳酸对葡萄糖得率也达到最大值,可作为指针病毒滴度及收毒时间的重要参数。

关键词： PK-15细胞; PPV病毒; 微载体; 反应器

Abstract:

In recent years, roller bottle was still used to produce porcine parvovirus. However, it was inefficient, labor-intensive and covered large areas. Now based on the optimization of time of infection (TOI), multiplicity of infection (MOI) and harvest time, porcine parvovirus static production process with PK-15 cells was well developed, that the maximum virus titre could reach 10^7.5 TCID₅₀/ml. More specifically, by optimizing TOI, the process was successfully transferred from static culture to microcarrier suspension culture, and 5L bioreactor verification experiment was conducted, which enable the maximum virus titer reached 10^7.2 TCID₅₀/ml. It was the first time to verify that the yield coefficients of lactate to glucose had a positive correlation with virus titre, that it could reach the peak value as soon as the porcine parvovirus at the maximum virus titre. It can be used as an important parameter for virus harvest.

Key words: PK-15 cells Porcine parvovirus Microcarrier Stirred bioreactor

收稿日期: 2015-04-02 出版日期: 2015-07-25

ZTFLH:

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通讯作者: 易小萍 E-mail: xpyi@ecust.edu.cn

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引用本文:

李智力, 易小萍, 储炬, 庄英萍, 张嗣良. PK-15细胞微载体悬浮培养生产猪细小病毒的工艺研究[J]. 中国生物工程杂志, 2015, 35(7): 62-67.

LI Zhi-li, YI Xiao-ping, CHU Ju, ZHUANG Ying-ping, ZHANG Si-liang. A Microcarrier Cell Culture Process for propagating Porcine Parvovirus in PK-15 Cells. China Biotechnology, 2015, 35(7): 62-67.

链接本文:

https://manu60.magtech.com.cn/biotech/CN/10.13523/j.cb.20150709 或 https://manu60.magtech.com.cn/biotech/CN/Y2015/V35/I7/62

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