• KSBB Journal
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• v.24 no.1
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• pp.1-8
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• 2009

In the past decade, we have observed rapid advances in the development of biochips in many fields including medical and environmental monitoring. Biochip experiments involve immobilizing a ligand on a solid substrate surface, and monitoring its interaction with an analyte in a sample solution. Metal nanoparticles can display extinction bands on their surfaces. These charge density oscillations are simply known as the localized surface plasmon resonance (LSPR). The high sensitivity of LSPR has been utilized to design biochips for the label-free detection of biomolecular interactions with various ligands. LSPR-based optical biochips and biosensors are easy to fabricate, and the apparatus cost for the evaluation of optical characteristics is lower than that for the conventional surface plasmon resonance apparatus. Furthermore, the operation procedure has become more convenient as it does not require labeling procedure. In this paper, we review the recent advances in LSPR research and also describe the LSPR-based optical biosensor constructed with a core-shell dielectric nanoparticle biochip for its application to label-free biomolecular detections such as antigen-antibody interaction.

• Korean Journal of Microbiology
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• v.54 no.4
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• pp.330-342
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• 2018

The Streptavidin and Biotin system has been studied most extensively as the high affinity non-covalent binding of Biotin to STR ($K_D=10^{-14}M$) and four Biotin binding sites in tetrameric Streptavidin makes this system useful for the production of multivalent antibody. For the application of this system, we cloned Streptavidin amplified from Streptomyces avidinii chromosome by PCR and fused to gene of hAY4 single-chain Fv antibody specific to death receptor 4 (DR4) which is a receptor for tumor necrosis factor ${\alpha}$ related apoptosis induced ligand. The hAY4 single-chain Fv antibody fused to Streptavidin expressed in Escherichia coli showed 43 kDa monomer in heated SDS-PAGE. However, this fusion protein shown in both non-heated SDS-PAGE and Size-exclusion chromatography exhibited 172 kDa as a tetramer suggesting that natural tetramerization of Streptavidin by non-covalent association induced hAY4 single-chain Fv tetramerization. This fusion protein retained a Biotin binding activity similar to natural Streptavidin as shown in Ouchterlony assay and ELISA. Death receptor 4 antigen binding activity of purified hAY4 single-chain Fv fused to Streptavidin was also confirmed by ELISA and Westernblot. In addition, surface plasmon resonance analysis showed 60-fold higher antigen binding affinity of the hAY4-STR than monomeric hAY4 ScFv due to tetramerization. In summary, hAY4 single-chain Fv fused to Streptavidin fusion protein was successfully expressed and purified as a soluble tetramer in E. coli and showed both Biotin and DR4 antigen binding activity suggesting possible production of bifunctional and tetrameric ScFv antibody.