Journal
APPLIED SCIENCE AND CONVERGENCE TECHNOLOGY
Volume 27, Issue 6, Pages 184-188Publisher
KOREAN VACUUM SOC
DOI: 10.5757/ASCT.2018.27.6.184
Keywords
Biosensor; Surface plasmon resonance; SPR; Toluene; Toluene binding domain
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Funding
- KRIBB Initiative Research Program (KRIBB, Korea)
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Herein, we report a novel surface plasmon resonance (SPR) sensor modified with a toluene-binding domain (TBD) to detect toluene. The TBD was manipulated with cysteines for rapid, easy, and orientation-controlled immobilization of the protein onto an SPR gold surface, due to the specific interaction between bare gold and the thiol group (-SH) of cysteines. After the purification and on-chip immobilization of the recombinant TBD, the thickness of cysteine-tagged TBD was determined to be approximately 1.8 nm according to atomic force microscopic cross-sectional analysis. The SPR analysis of cysteine-tagged TBD adsorption showed that cysteine-tagged TBD binds to gold thin film with high binding affinity. With an SPR-based ligand binding assay, a detectable increase in the SPR response was observed for 15.62 mu M of toluene. Owing to the inherent specificity of the TBD, this method showed comparative specificity for the determination of toluene, as it exhibited no SPR response to aromatic compounds such as p-xylene or benzene. Taken together, the results obtained from this study indicate that a TBD-modified SPR sensor system can be useful for detecting toluene, a volatile organic compound widely distributed in the environment.
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