Protein nanopatterns and biosensors using gold binding polypeptide as a fusion partner.
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.
Park
Tae Jung
TJ
Metabolic and Biomolecular Engineering National Research Laboratory, BioProcess Engineering Research Center, Korea Advanced Institute of Science and Technology (KAIST), Daejeon 305-701, Korea.
Lee
Sang Yup
SY
Lee
Seok Jae
SJ
Park
Jong Pil
JP
Yang
Kwang Suk
KS
Lee
Kyung-Bok
KB
Ko
Sungho
S
Park
Jong Bae
JB
Kim
Taekeun
T
Kim
Seong Kyu
SK
Shin
Yong Bum
YB
Chung
Bong Hyun
BH
Ku
Su-Jin
SJ
Kim
Do Hyun
DH
Choi
Insung S
IS
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*
D003102:Colloids; D010455:Peptides; D011993:Recombinant Fusion Proteins; D006046:Gold; D004247:DNA