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优化蛋白表达并建立用于杆状病毒/昆虫细胞培养的波浪生物反应器。

Optimisation of protein expression and establishment of the Wave Bioreactor for Baculovirus/insect cell culture.

机构信息

Core Technologies/Biomolecules Production, Novartis Pharma AG, Research, Building WSJ-506.101, CH-4002, Basel, Switzerland.

出版信息

Cytotechnology. 2002 Jan;38(1-3):77-85. doi: 10.1023/A:1021102015070.

Abstract

As the interest of research is beginning to shift from genomicsto proteomics the number of proteins to be expressed is rapidlyincreasing. To do so, well-established, high-level expressionsystems and rapid, cost-effective production means are needed. For addressing the latter, a novel cultivation system for recombinant cells, the Wave Bioreactortrade mark has recently becomeavailable. We describe the set-up and the optimisation of parameters essential for successful operation and growth of insect cells to high cell densities in the Wave Bioreactor. According to our experience, the Cellbagtrade mark system comparesvery favorably to conventional cultivation vessels such as bioreactors and roller cultures with respect to simplicity ofoperation and cost. Additionally, we developed a rapid and simple protocol for assessing expression and production conditions for the Baculovirus/insect cell system applicable to many different genes/proteins. Important parameters like MOI,TOI, peak cell density (PCD) and expression levels are determinedin pre-experiments on small scale to achieve optimal expressionof a given protein. These conditions are subsequently transformedand applied to large scale cultures grown in nutrient-supplemented medium in the Wave Bioreactor.

摘要

随着研究的兴趣从基因组学转向蛋白质组学,需要表达的蛋白质数量正在迅速增加。为此,需要建立完善的、高水平的表达系统和快速、具有成本效益的生产手段。为了解决后者问题,最近出现了一种新型的重组细胞培养系统——Wave 生物反应器商标。我们描述了 Wave 生物反应器中昆虫细胞成功操作和生长至高密度所必需的设置和参数的优化。根据我们的经验,Cellbag 商标系统在操作简单性和成本方面与传统的培养容器(如生物反应器和滚瓶培养)相比具有很大的优势。此外,我们开发了一种快速简单的 Baculovirus/昆虫细胞系统表达和生产条件评估方法,适用于许多不同的基因/蛋白质。在小规模预实验中确定重要参数,如 MOI、TOI、峰值细胞密度(PCD)和表达水平,以实现给定蛋白质的最佳表达。然后将这些条件转化并应用于在 Wave 生物反应器中用营养补充培养基培养的大规模培养中。

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