The Phenylboronic acid Derivative as linker for Surface Modification of QCM Chip and its Application on Dengue virus Detection in Human Serum

• Main Authors: He, Jian-Xing, 何建興
• Other Authors: Li, Yaw-Kuen
• Format: Others
• Language: zh-TW
• Published: 2012
• Online Access: http://ndltd.ncl.edu.tw/handle/t9atx2

碩士 === 國立交通大學 === 應用化學系分子科學碩博士班 === 101 === The Quartz crystal microbalance (QCM) is a useful platform for detecting the interactions between bio-molecules such as antigen/antibody, aptamer/protein…etc. For promoting the detection sensitivity of QCM and reduce the cost from chip comsumption, we synthesized the interface linker by coupling the aminophenylboronate with the carboxylic residue of the 2-thiolpyridine-capped 11-mercaptoundecanoic acid. The phenylboronic acid residue was used for controlling the orientation of antibody on chip through the complex formation of boronic acid with sugar moiety on antibody. In order to establish the chip regeneration process, ethylene dithiol was coated on gold chip surface through the self-assembly manner. The thiol on chip surface was then employed to replace the thiolpyridine moiety of the linker to form a disulfide bond, which can be broken down by treating with tris(2-carboxyethyl)phosphine. The experimental results revealed that the sensor chip can be reused, with minor loss of sensitivity, for at least 4 times. Quantitative assessment on Dengue virus 1 (DV1) detection was performed using this new strategy. 1% N-laurylsarcosine (LSS) was further added in a serum sample (with 100-fold dilution) to eliminate non-specific interaction of chip and molecules in serum. Results showed that the detection exhibited a good linear co-relation in the range of 1000 to 1,000,000 cfu/ml of DV1.