Real-Time Detection of α-Thrombin Binding to Single-Strand DNA Aptamers and Dengue Virus DNA Hybridization by ARROW-B SPR Biosensors

• Main Authors: Chi-Chieh Huang, 黃繼傑
• Other Authors: Yang-Tung Huang
• Format: Others
• Language: en_US
• Published: 2008
• Online Access: http://ndltd.ncl.edu.tw/handle/73575633967046751075

碩士 === 國立交通大學 === 電子工程系所 === 96 === In this thesis, an antiresonant reflecting optical waveguide of type B (ARROW-B) surface plasmon resonance (SPR) biosensor operating in the aqueous environment has been investigated. The ARROW-B SPR biosensor is proposed to provide a label-free, high-throughput and highly surface-sensitive platform to detect the bimolecular interactions in real time. The design and fabrication process of the ARROW-B SPR sensor chips are described and discussed. Besides, the primary analytes for the bioassay experiments are divided into two categories based on different binding characteristics. First, the real-time detection of α-thrombin binding to ssDNA aptamers was under in-depth investigation. The gold nanoparticles modified with anti-thrombin antibodies were employed to bind to the α-thrombins for signal amplification. The detection limit of this biosensor to α-thrombin was measured at 1 pM level, which was comparable to that of the Biacore 3000 system but at much lower cost. Second, the real-time detection of dengue virus ssDNA hybridization was studied. The dengue virus DNA probe was modified with a thiol group at one end to achieve effective immobilization on the Au surface, while the DNA target utilized the complementary sequence to bind to the immobilized probe. In summary, the measurement results have shown that the ARROW-B SPR biosensors can be applied to detect the ssDNA aptamer/α-thrombin interaction and dengue virus ssDNA hybridization both quantitatively and qualitatively in real time.