Davies Angela, Greene April, Lullau Elke, Abbott W Mark
AstraZeneca, Mereside, Alderley Park, Macclesfield, Cheshire, UK.
Protein Expr Purif. 2005 Jul;42(1):111-21. doi: 10.1016/j.pep.2005.03.012. Epub 2005 Mar 31.
Transient transfection of mammalian cells with episomal vectors is a very useful method for producing high levels of recombinant proteins. Transient systems remove the need for the laborious and time-consuming process of creating stable cell lines. Here, we describe the optimisation and evaluation of a high-throughput transient expression system in HEK293-EBNA cells. The process was developed for the expression of 10 constructs simultaneously in deep-well plates and subsequent purification using 96-well plate affinity chromatography. This enabled multiple combinations of different constructs, vectors, and expression conditions to be studied in parallel.
用游离型载体对哺乳动物细胞进行瞬时转染是生产高水平重组蛋白的一种非常有用的方法。瞬时系统省去了创建稳定细胞系这一费力且耗时的过程。在此，我们描述了HEK293-EBNA细胞中高通量瞬时表达系统的优化和评估。该方法是为在深孔板中同时表达10种构建体并随后使用96孔板亲和色谱法进行纯化而开发的。这使得能够并行研究不同构建体、载体和表达条件的多种组合。
