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

• Published in: Analytical chemistry (Washington) Vol. 78; no. 20; pp. 7197 - 7205
• Main Authors: Park, Tae Jung, Lee, Sang Yup, Lee, Seok Jae, Park, Jong Pil, Yang, Kwang Suk, Lee, Kyung-Bok, Ko, Sungho, Park, Jong Bae, Kim, Taekeun, Kim, Seong Kyu, Shin, Yong Bum, Chung, Bong Hyun, Ku, Su-Jin, Kim, Do Hyun, Choi, Insung S
• Format: Journal Article
• Language: English
• Published: Washington, DC American Chemical Society 15.10.2006
• Subjects: Analytical chemistry; Binding sites; Biological and medical sciences; Biological Assay; Biosensors; Biotechnology; Chemistry; Colloids; Deoxyribonucleic acid; DNA; DNA - chemistry; DNA - genetics; Exact sciences and technology; Fundamental and applied biological sciences. Psychology; Genes, Reporter - genetics; Gold; Gold - chemistry; Methods. Procedures. Technologies; Microscopy, Atomic Force; Nanostructures - chemistry; Peptides; Peptides - chemistry; Polymerase Chain Reaction; Proteins; Recombinant Fusion Proteins - chemistry; Recombinant Fusion Proteins - genetics; Recombinant Fusion Proteins - metabolism; Salmonella; SARS coronavirus; Severe acute respiratory syndrome; Spectrometric and optical methods; Streptomyces; Surface Plasmon Resonance - instrumentation; Surface Plasmon Resonance - methods; Various methods and equipments; Atomic force microscopy; Gold; Antibody; Nanoparticle; Immobilization; Protein; Absorption spectrometry; Chemical modification; Polypeptide; Biosensor; DNA; Surface plasmon resonance; DNA hybridization; Microfluidics; Ultraviolet visible spectrometry
• ISSN: 0003-2700; 1520-6882
• DOI: 10.1021/ac060976f

An efficient strategy for immobilizing proteins on a gold surface was developed by employing the gold binding polypeptide (GBP) as a fusion partner. Using the enhanced green fluorescent protein (EGFP), severe acute respiratory syndrome coronavirus (SARS-CoV) envelope protein (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 bare gold surface were demonstrated. The GBP-fused SCVme bound to gold nanoparticles successfully interacted with its antibody and showed changes in absorbance and color, allowing efficient diagnosis of SARS-CoV. The fusion proteins could be successfully immobilized on the gold surface by nanopatterning and microcontact printing as examined by atomic force microscopy and surface plasmon resonance analysis. The poly(dimethylsiloxane) microfluidic channels were created on the gold surface and were used for antigen−antibody and DNA−DNA interaction studies. Specific immobilization of GBP−EGFP fusion protein and its interaction with the antibody in the microchannels could be demonstrated. By immobilizing the DNA probe through the use of GBP-fused cSA, specific hybridization of the target DNA prepared from Salmonella could also be achieved. The GBP-fusion method allows immobilization of proteins onto the gold surface without surface modification and in bioactive forms suitable for studying protein−protein, DNA−DNA, and other biomolecular interaction studies. Furthermore, these studies can be carried out in a microfluidic system, which allows high-throughput analysis of biomolecular interactions.