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Improvements to the throughput of recombinant protein expression in the baculovirus/insect cell system

机译：改进杆状病毒/昆虫细胞系统中重组蛋白表达的通量

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Recombinant baculoviruses have proved to be a very useful means to express many proteins over the last 20 years. Since their introduction, there have been a number of significant improvements that have simplified and speeded up the construction of baculoviruses. One of the most commonly used methods relies upon recombination with the baculovirus genome maintained in Escherichia coli. In this paper, we report the conversion of nearly all the steps in this process including the expression testing and purification to a multi-well plate format. This enables a significant increase in the number of constructs that can be processed in a shorter period of time and an order of magnitude increase in the number of expression conditions that can be analysed. A key step in our process is that the transfection is done in suspension rather than adherent cells, which gives a much higher virus titre than in the standard methods. (c) 2005 Elsevier Inc. All rights reserved.
机译：在过去的20年中，重组杆状病毒被证明是表达许多蛋白质的非常有用的手段。自引入以来，已经进行了许多重大改进，简化并加快了杆状病毒的构建。最常用的方法之一是与保持在大肠杆菌中的杆状病毒基因组重组。在本文中，我们报告了该过程中几乎所有步骤的转化，包括表达测试和纯化为多孔板形式。这使得可以在较短的时间段内处理的构建体的数量显着增加，并且可以分析的表达条件的数量增加一个数量级。我们过程中的关键步骤是转染是在悬浮液中而不是在贴壁细胞中进行，与标准方法相比，这种方法的病毒滴度要高得多。 （c）2005 Elsevier Inc.保留所有权利。

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