Protein Nanopatterns and Biosensors Using Gold Binding Polypeptide as a Fusion Partner

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• 作者：Tae Jung Park ; Sang Yup Lee ; Seok Jae Lee ; Jong Pil Park ; Kwang Suk Yang ; Kyung-Bok Lee ; Sungho Ko ; Jong Bae Park ; Taekeun Kim ; Seong Kyu Kim ; Yong Bum Shin ; Bong Hyun Chung ; Su-Jin Ku ; Do Hyun Kim ; In
• 刊名：Analytical Chemistry
• 出版年：2006
• 出版时间：October 15, 2006
• 年：2006
• 卷：78
• 期：20
• 页码：7197 - 7205
• 全文大小：617K
• 年卷期：v.78,no.20(October 15, 2006)
• ISSN：1520-6882

文摘

An efficient strategy for immobilizing proteins on a goldsurface was developed by employing the gold bindingpolypeptide (GBP) as a fusion partner. Using the enhanced green fluorescent protein (EGFP), severe acuterespiratory syndrome coronavirus (SARS-CoV) envelopeprotein (SCVme), and core streptavidin (cSA) of Streptomyces avidinii as model proteins, specific immobilization of the GBP-fusion proteins onto the gold nanoparticles and generation of protein nanopatterns on the baregold surface were demonstrated. The GBP-fused SCVmebound to gold nanoparticles successfully interacted withits antibody and showed changes in absorbance and color,allowing efficient diagnosis of SARS-CoV. The fusionproteins could be successfully immobilized on the goldsurface by nanopatterning and microcontact printing asexamined by atomic force microscopy and surface plasmon resonance analysis. The poly(dimethylsiloxane) microfluidic channels were created on the gold surface andwere used for antigen-antibody and DNA-DNA interaction studies. Specific immobilization of GBP-EGFP fusion protein and its interaction with the antibody in themicrochannels could be demonstrated. By immobilizingthe DNA probe through the use of GBP-fused cSA, specifichybridization of the target DNA prepared from Salmonella could also be achieved. The GBP-fusion methodallows immobilization of proteins onto the gold surfacewithout surface modification and in bioactive forms suitable for studying protein-protein, DNA-DNA, and otherbiomolecular interaction studies. Furthermore, thesestudies can be carried out in a microfluidic system, whichallows high-throughput analysis of biomolecular interactions.