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

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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.

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Abbreviations

GST–GFP:

Glutathione S-transferase-fused green fluorescent protein

IPTG:

Isopropyl β-d-thiogalactopyranoside

PDMS:

Poly-dimethylsiloxane

SDS–PAGE:

Sodium dodecyl sulfate-polyacrylamide gel electrophoresis

SPR:

Imaging surface plasmon resonance imaging

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Acknowledgments

We thank Dr. Choong-Min Ryu (KRIBB) for his helpful comments on this manuscript. This research was supported by grants from the KRIBB Initiative Research Program (KRIBB, Korea), the Protein Chip Technology Program, and the Nano/Bio Science and Technology Program (MOST, Korea).

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Authors and Affiliations

  1. BioNanotechnology Research Center, Korea Research Institute of Bioscience and Biotechnology, 115, Yuseong, Daejeon, 305–600, South Korea

    Moonil Kim, So-Young Lee, Hyunju Choi, Yong-Beom Shin, Sun Ok Jung, Min-Gon Kim & Bong Hyun Chung

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  1. Moonil Kim
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  2. So-Young Lee
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  3. Hyunju Choi
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  4. Yong-Beom Shin
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  5. Sun Ok Jung
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  6. Min-Gon Kim
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  7. Bong Hyun Chung
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Corresponding author

Correspondence to Bong Hyun Chung.

Additional information

M. Kim and S. Y. Lee contributed equally to this work.

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Kim, M., Lee, SY., Choi, H. et al. On-chip Escherichia coli culture, purification, and detection of expressed proteins. Eur Biophys J 35, 655–662 (2006). https://doi.org/10.1007/s00249-006-0072-8

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  • DOI: https://doi.org/10.1007/s00249-006-0072-8

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