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

Cited 11 time in scopus
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Title
On-chip Escherichia coli culture, purification, and detection of expressed proteins
Author(s)
Moonil Kim; So Young Lee; Hyun-Ju Choi; Yong Beom Shin; Sun Ok Jung; Min-Gon Kim; Bong Hyun Chung
Bibliographic Citation
European Biophysics Journal with Biophysics Letters, vol. 35, no. 8, pp. 655-662
Publication Year
2006
Abstract
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.
Keyword
Glutathione S-transferase-fused green fluorescent protein (GST-GFP)Microwell chipOn-chip purificationSurface plasmon resonance (SPR) imaging system
ISSN
0175-7571
Publisher
Springer
DOI
http://dx.doi.org/10.1007/s00249-006-0072-8
Type
Article
Appears in Collections:
Critical Diseases Diagnostics Convergence Research Center > 1. Journal Articles
Division of Research on National Challenges > Bionanotechnology Research Center > 1. Journal Articles
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