Analysis of recombinant protein expression using localized surface plasmon resonance (LSPR).
The localized surface plasmon resonance (LSPR)-based optical biosensor using nano-structures of noble metals has been considered as a useful tool for label-free detection of DNA hybridization and protein-protein interactions. We fabricated LSPR-based optical biosensors using gold nano-islands (nominal thickness; 75 A) on glass substrates that were easily made using the conventional fabrication methods. The formation of gold nano-islands on glass substrates was realized by heat treatment of thin gold film deposited with a low deposition rate (approximately 0.05 A/s). The morphologies of sensor surfaces composed of gold nano-islands were observed using an atomic force microscope (AFM) with a non-contact mode. To investigate the sensing capacity of the gold nano-island sensor for the binding of proteins by affinity interactions, the streptavidin and biotin interaction was used as a model system. In addition, detection of recombinant glutathione-S-transferase (GST)-tagged human interleukin-6 (hIL6) expressed in Escherichia coli was carried out by LSPR. It is expected that the LSPR sensors composed of gold nano-islands can be an alternative to traditional methods such as SDS-polyacrylamide gel electrophoresis (SDS-PAGE) for fast analysis of protein expression.
Shin
Yong-Beom
YB
BioNanotechnology Research Center, Korea Research Institute of Bioscience and Biotechnology, Yusong-gu, Daejeon 305-333, Republic of Korea. ybshin@kribb.re.kr
Lee
Jeong-Min
JM
Park
Mi-Ra
MR
Kim
Min-Gon
MG
Chung
Bong Hyun
BH
Pyo
Hyeon-Bong
HB
Maeng
Sunglyul
S
D003001:Cloning, Molecular; D004926:Escherichia coli; D006046:Gold; D006801:Humans; D015850:Interleukin-6 / Q000032:analysis / Q000096:biosynthesis / Q000235:genetics; D011994:Recombinant Proteins / Q000032:analysis* / Q000096:biosynthesis / Q000235:genetics; D020349:Surface Plasmon Resonance
D015850:Interleukin-6; D011994:Recombinant Proteins; D006046:Gold