title: Protein nanopatterns and biosensors using gold binding polypeptide as a fusion partner.
abstract: 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.
authors.affiliation: Metabolic and Biomolecular Engineering National Research Laboratory, BioProcess Engineering Research Center, Korea Advanced Institute of Science and Technology (KAIST), Daejeon 305-701, Korea.
authors.forename: Tae Jung
authors.initials: TJ
authors.lastname: Park
authors.forename: Sang Yup
authors.initials: SY
authors.lastname: Lee
authors.forename: Seok Jae
authors.initials: SJ
authors.lastname: Lee
authors.forename: Jong Pil
authors.initials: JP
authors.lastname: Park
authors.forename: Kwang Suk
authors.initials: KS
authors.lastname: Yang
authors.forename: Kyung-Bok
authors.initials: KB
authors.lastname: Lee
authors.forename: Sungho
authors.initials: S
authors.lastname: Ko
authors.forename: Jong Bae
authors.initials: JB
authors.lastname: Park
authors.forename: Taekeun
authors.initials: T
authors.lastname: Kim
authors.forename: Seong Kyu
authors.initials: SK
authors.lastname: Kim
authors.forename: Yong Bum
authors.initials: YB
authors.lastname: Shin
authors.forename: Bong Hyun
authors.initials: BH
authors.lastname: Chung
authors.forename: Su-Jin
authors.initials: SJ
authors.lastname: Ku
authors.forename: Do Hyun
authors.initials: DH
authors.lastname: Kim
authors.forename: Insung S
authors.initials: IS
authors.lastname: Choi
mesh_terms: D001681:Biological Assay; D003102:Colloids; D004247:DNA / Q000737:chemistry / Q000235:genetics; D017930:Genes, Reporter / Q000235:genetics; D006046:Gold / Q000737:chemistry; D018625:Microscopy, Atomic Force; D049329:Nanostructures / Q000737:chemistry*; D010455:Peptides / Q000737:chemistry*; D016133:Polymerase Chain Reaction; D011993:Recombinant Fusion Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D020349:Surface Plasmon Resonance / Q000295:instrumentation* / Q000379:methods*
chemical_list: D003102:Colloids; D010455:Peptides; D011993:Recombinant Fusion Proteins; D006046:Gold; D004247:DNA