高通量灌流培养模型在生物工艺开发中的应用研究

doi:10.13523/j.cb.2005005

中国生物工程杂志

2020, Vol. 40

Issue (8): 63-73 DOI: 10.13523/j.cb.2005005

综述

高通量灌流培养模型在生物工艺开发中的应用研究

靳露¹,周航^2,^*,曹云²,王振守²,曹荣月^1,^*(

)

1 中国药科大学生命科学与技术学院南京 211198
2 上海药明生物技术有限公司上海 200131

Research on Applications of High-Throughput Perfusion Models in Bioprocessing Development

JIN Lu¹,ZHOU Hang^2,^*,CAO Yun²,WANG Zhou-shou²,CAO Rong-yue^1,^*(

)

1 College of Life Science and Biotechnology,China Pharmaceutical University,Nanjing　211198,China
2 Cell Culture Process Development,Wuxi Biologics,Shanghai　200131,China

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摘要：

近年来,连续型细胞培养由于其高单位体积产量、稳定的产品质量属性以及潜在的成本节约效应正成为生物大分子制药生产的工艺焦点。相比传统的流加培养模式,灌流培养因培养的连续性、操作的复杂性,致使其反应器规模培养需消耗大量培养基,产生更高人力成本,不能满足当今加速化高效化的工艺开发需求。为获得稳健的灌流培养工艺并控制较低成本,高通量灌流培养模型被用于批量化的小规模灌流培养,进行灌流培养前期的克隆筛选、培养基筛选及工艺参数优化等工作,为后期大规模培养提供实用性数据支持,同时也被用于预测大规模培养的细胞表型和产品质量属性。重点介绍了当前高通量系统包括摇瓶/摇管系统、多平行自动化系统以及微流控体系用作灌流培养的特征、具体应用及比较,同时论述当前高通量灌流培养系统在生物工艺领域发展所面临的机遇及挑战,并展望其应用前景。

关键词： 灌流培养; 高通量; 微型生物反应器; Ambr; 微流控

Abstract:

Recently, continuous cell culture is becoming the process focus in the pharmaceutical industry due to its high volumetric productivity, stable product quality attributes, and cost- effectiveness. Compared to the traditional fed-batch culture, benchtop-scale perfusion culture requires quantities of media and labor costs due to its longer culture duration and operation complexity, thus failing to satisfy the current requirement of accelerated and efficient process development. To obtain a robust perfusion process with reduced costs, high-throughput perfusion models are utilized for batches of small-scale perfusion culture in the early-stage process development including clone screening, media selection and process parameter optimization, providing practical process data for late-stage large-scale bioprocessing. Furthermore, they are also applied to predict the phenotype and product quality attributes in large-scale culture. This article will focus on the characteristics, applications and comparisons of current high-throughput systems including shake flasks and spin tubes, parallelized automated ambr systems and microfluidic systems, and discuss the opportunities and challenges faced with high-throughput perfusion models in the bioprocessing development, then look forward to the future prospects.

Key words: Perfusion High-throughput Micro-bioreactor Ambr Microfluidics

收稿日期: 2020-05-06 出版日期: 2020-09-10

ZTFLH:

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通讯作者: 周航,曹荣月 E-mail: caorongyuenanjing@126.com

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引用本文:

靳露,周航,曹云,王振守,曹荣月. 高通量灌流培养模型在生物工艺开发中的应用研究[J]. 中国生物工程杂志, 2020, 40(8): 63-73.

JIN Lu,ZHOU Hang,CAO Yun,WANG Zhou-shou,CAO Rong-yue. Research on Applications of High-Throughput Perfusion Models in Bioprocessing Development. China Biotechnology, 2020, 40(8): 63-73.

链接本文:

https://manu60.magtech.com.cn/biotech/CN/10.13523/j.cb.2005005 或 https://manu60.magtech.com.cn/biotech/CN/Y2020/V40/I8/63

图1 反应器灌流培养装置图

图2 摇管灌流操作流程[28]

图3 Ambr? 15工作站构造①(①图片来源:www.sartorius.com.cn)

图4 Ambr? 250灌流培养系统构造①(①图片来源:www.sartorius.com.cn)

图5 微流控技术在细胞培养工艺中的应用示例

表1 高通量灌流培养模型比较

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