Optimisation and evaluation of a high-throughput mammalian protein expression system

被引：18

作者：

Davies, A

Greene, A

Lullau, E

Abbott, WM

机构：

[1] AstraZeneca, Macclesfield SK10 4TG, Cheshire, England

[2] AstraZeneca R&D, S-15185 Sodertalje, Sweden

来源：

PROTEIN EXPRESSION AND PURIFICATION | 2005年 / 42卷 / 01期

关键词：

high-throughput expression; heterologous expression; recombinant protein; HEK293-EBNA cells; episomal vector; mammalian expression;

D O I：

10.1016/j.pep.2005.03.012

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

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. (c) 2005 Elsevier Inc. All rights reserved.

引用

页码：111 / 121

页数：11

共 22 条

[1]

ABREMSKI K, 1984, J BIOL CHEM, V259, P1509

[2] High-throughput screening for soluble recombinant expressed kinases in Escherichia coli and insect cells [J].