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PubMed · 15203330

A fusion protein expression analysis using surface plasmon resonance imaging.

Jin-Mi Jung·Yong-Beom Shin·Min-Gon Kim·Hyeon-Su Ro·Hee-Tae Jung·Bong Hyun Chung
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Abstract

A surface plasmon resonance (SPR) imaging system was constructed and used to detect the affinity-tagged recombinant proteins expressed in Escherichia coli. With regards to model proteins, the hexahistidine-ubiquitin-tagged human growth hormone (His(6)-Ub-hGH), glutathione S-transferase-tagged human interleukin-6 (GST-hIL6), and maltose-binding protein-tagged human interleukin-6 (MBP-hIL6) expressed in E. coli were analyzed. The cell lysates were spotted on gold thin films coated with 11-mercaptoundecanol (MUOH)/dextran derivatized with Ni(II)-iminodiacetic acid (IDA-Ni(II)), glutathione, or cyclodextrin. After a brief washing of the gold chip, SPR imaging measurements were carried out in order to detect the bound affinity-tagged fusion proteins. Using this new approach, rapid high-throughput expression analysis of the affinity-tagged proteins were obtained. The SPR imaging protein chip system used to measure the expression of affinity-tagged proteins in a high-throughput manner is expected to be an attractive alternative to traditional laborious and time-consuming methods, such as SDS-polyacrylamide gel electrophoresis (SDS-PAGE) and Western blots.

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Affinity LabelsCarrier ProteinsCloning, MolecularEscherichia coliFatty AlcoholsGene ExpressionGlutathione TransferaseGrowth HormoneHistidineHumansInterleukin-6Maltose-Binding ProteinsRecombinant Fusion ProteinsSulfhydryl CompoundsSurface Plasmon ResonanceUbiquitinJournal ArticleResearch Support, Non-U.S. Gov't

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Jin-Mi Jung, Yong-Beom Shin, Min-Gon Kim, Hyeon-Su Ro, Hee-Tae Jung, Bong Hyun Chung. 2004-07-15. A fusion protein expression analysis using surface plasmon resonance imaging.. https://doi.org/10.1016/j.ab.2004.02.009

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