Biotechnology and Bioprocess Engineering:BBE

The Korean Society for Biotechnology and Bioengineering (한국생물공학회)
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Polyhydroxyalkanoate Chip for the Specific Immobilization of Recombinant Proteins and Its Applications in Immunodiagnostics

  • Park, Tae-Jung (Department of Chemical and Biomolecular Engineering, Bioprocess Engineering Research Center, Center for Ultramicrochemical Process Systems, Korea Advanced Institute of Science and Technology) ;
  • Park, Jong-Pil (Department of Chemical and Biomolecular Engineering, Bioprocess Engineering Research Center, Center for Ultramicrochemical Process Systems, Korea Advanced Institute of Science and Technology) ;
  • Lee, Seok-Jae (Department of Chemical and Biomolecular Engineering, Bioprocess Engineering Research Center, Center for Ultramicrochemical Process Systems, Korea Advanced Institute of Science and Technology) ;
  • Hong, Hyo-Jeong (The Antibody Engineering Laboratory, Korea Research Institute of Bioscience and Biotechnology) ;
  • Lee, Sang-Yup (Department of Chemical and Biomolecular Engineering, Bioprocess Engineering Research Center, Center for Ultramicrochemical Process Systems, Korea Advanced Institute of Science and Technology)
  • Published : 2006.04.30
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Abstract

In this study, a novel strategy was developed for the highly selective immobilization of proteins, using the polyhydroxyalkanoate (PHA) depolymerase substrate binding domain (SBD) as an active binding domain. In order to determine the appropriacy of this method for immunodiagnostic assays, the single-chain antibody (ScFv) against the hepatitis B virus (HBV) preS2 surface protein and the severe acute respiratory syndrome coronavirus (SARS-CoV) envelope protein (SCVe) were fused to the SBD, then directly immobilized on PH A-coated slides via microspotting. The fluorescence-labeled HBV antigen and the antibody against SCVe were then utilized to examine specific interactions on the PHA-coated surfaces. Fluorescence signals were detected only at the spotted positions, thereby indicating a high degree of affinity and selectivity for their corresponding antigens/antibodies. Furthermore, we detected small amounts of ScFv-SBD (2.7 ng/mL) and SCVe-SBD fusion proteins (0.6ng/mL). Therefore, this microarray platform technology, using PHA and SBD, appears generally appropriate for immunodiagnosis, with no special requirements with regard to synthetic or chemical modification of the biomolecules or the solid surface.

Keywords

References

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