首页 | 本学科首页   官方微博 | 高级检索  
     


Optimisation of protein expression and establishment of the Wave Bioreactor for Baculovirus/insect cell culture
Authors:Wilfried Weber  Eric Weber  Sabine Geisse  Klaus Memmert
Affiliation:(1) Core Technologies/Biomolecules Production, Novartis Pharma AG, Research, Building WSJ-506.101, CH-4002 Basel, Switzerland;(2) Present address: ETH Hoenggerberg, CH-8093 Zürich, Switzerland
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 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 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.
Keywords:baculovirus/insect cell system  essential parameters for operation  expression optimisation  feeding strategies  Wave Bioreactor
本文献已被 SpringerLink 等数据库收录!
设为首页 | 免责声明 | 关于勤云 | 加入收藏

Copyright©北京勤云科技发展有限公司  京ICP备09084417号