On-chip Escherichia coli culture, purification, and detection of expressed proteins

被引:11
作者
Kim, Moonil [1 ]
Lee, So-Young [1 ]
Choi, Hyunju [1 ]
Shin, Yong-Beom [1 ]
Jung, Sun Ok [1 ]
Kim, Min-Gon [1 ]
Chung, Bong Hyun [1 ]
机构
[1] BioNanotechnol Res Ctr, Korea Res Inst Biosci & Biotechnol, Taejon 305600, South Korea
来源
EUROPEAN BIOPHYSICS JOURNAL WITH BIOPHYSICS LETTERS | 2006年 / 35卷 / 08期
关键词
microwell chip; surface plasmon resonance (SPR) imaging system; glutathione S-transferase-fused green fluorescent protein (GST-GFP); on-chip purification;
D O I
10.1007/s00249-006-0072-8
中图分类号
Q6 [生物物理学];
学科分类号
071011 ;
摘要
In a recent study, we reported the results of a rapid high-throughput expression analysis of the affinity-tagged proteins present in total cell lysates, using a surface plasmon resonance (SPR) imaging protein chip system. In this paper, we describe a novel method, which is able to sequentially carry out a recombinant Escherichia coli culture, as well as the detection and purification of the expressed proteins on a single microwell chip, fabricated on a two-dimensional thin gold film. Following the induction of the protein on the microwell chip, the E. coli cells were lysed on the chip via the addition of lysozymes, and the expressed glutathione S-transferase-fused green fluorescent protein (GST-GFP) was then purified on the chip via affinity interaction with the glutathionylated gold surface of the chip. Finally, the expressed protein was directly detected using the surface plasmon resonance (SPR) imaging system. This system saves a substantial amount of time, experimental resources, and labor, by allowing for the complicated and labor-intensive procedures inherent to the production of recombinant proteins to be conducted on a single microwell chip, simply and economically.
引用
收藏
页码:655 / 662
页数:8
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