Biotechnology and Bioprocess Engineering:BBE

The Korean Society for Biotechnology and Bioengineering (한국생물공학회)
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Surface Plasmon Resonance Imaging Analysis of Hexahistidine-tagged Protein on the Gold Thin Film Coated with a Calix Crown Derivative

  • Chung, Bong-Hyun (BioNanotechnology Research Center, Korea Research Institute of Bioscience and Biotechnology) ;
  • Baek, Seung-Hak (BioNanotechnology Research Center, Korea Research Institute of Bioscience and Biotechnology, Departmet of Biological Engineering, Inha University) ;
  • Shin, Yong-Beom (BioNanotechnology Research Center, Korea Research Institute of Bioscience and Biotechnology) ;
  • Kim, Min-Gon (BioNanotechnology Research Center, Korea Research Institute of Bioscience and Biotechnology) ;
  • Ro, Hyeon-Su (BioNanotechnology Research Center, Korea Research Institute of Bioscience and Biotechnology) ;
  • Kim, Eun-Ki (Departmet of Biological Engineering, Inha University)
  • Published : 2004.03.01
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Abstract

A surface plasmon resonance (SPR) imaging system was constructed and used to detect the hexahistidine-ubiquitin-tagged human parathyroid hormone fragment (His$\sub$6/-Ub-hPTHF(1-34)) expressed in Escherichia coli. The hexahistidine-specific antibody was immobilized on a thin gold film coated with ProLinker$\^$TM/ B, a novel calixcrown derivative with a bifunctional coupling property that permits efficient immobilizaton of capture proteins on solid matrices. The soluble and insoluble fractions of an E. coli cell lysate were spotted onto the antibody-coated gold chip, which was then washed with buffer (pH 7.4) solution and dried. SPR imaging measurements were carried out to detect the expressed His$\sub$6/-Ub-hPTHF(1-34). There was no discernible protein image in the uninduced cell lysate, indicating that non-specific binding of contaminant proteins did not occur on the gold chip surface. It is expected that the approach used here to detect affinity-tagged recombinant proteins using an SPR imaging technique could be used as a powerful tool for the analyses of a number of proteins in a high-throughput mode.

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References

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