光纖式奈米生物感測器結合高效能液相層析串聯質譜儀分析系統開發與方法驗證

• Main Authors: Lin, Ching-Yi, 林靜宜
• Other Authors: Wang, Shau-Chun
• Format: Others
• Language: zh-TW
• Published: 2015
• Online Access: http://ndltd.ncl.edu.tw/handle/17639228416814200219

碩士 === 國立中正大學 === 化學暨生物化學研究所 === 103 === There have been considerable numbers of publications regarding connecting the mass spectrometer and the surface plasmon resonance biosensors. This integrated system combines the biosensors’ benefits of affinity capture and mass spectrometry sensitivity to identify analytes . In this study, the integration of high-performance liquid chromatography electrospray tandem mass spectrometry with the label-free fiber-optic particle plasmon resonance biosensor (FO-PPR) is investigated. Unlike using MALDI-TOF-MS analysis of complex protein, of which the disadvantages include matrix inhomegeneity sample loss, using various mobile phase solvents of different polarity, analytes are able to elute and separate with the chromatographic column prior to determining with the mass spectrometer, when matrix interferences are removed. Because of the advantages of label-free, real-time detection, high sensitivity, and specificity, FO-PPR is suitable to detect biochemical molecules, such as: the interaction of antigen-antibody proteins . In this study, two protein detection methods, using FO-PPR and LC-MS / MS respectively, are compared to evaluate their sensitivity with protein myoglobin and ovalbumin (OVA) , In addition desorbed analytes from FO- PPR fibers are injected into LC-MS/MS to evaluate their recovery, for finding out the proper operating conditions of FO-PPR and LC-MS / MS integration system. The results suggest that: (1) For detection of myoglobin, LC-MS had a good linear for concentration of 5 ~ 50 μg / ml, FO-PPR had a good linear relationship on the concentration of 50-1000 μg / ml . Detecting OVA, LC-MS system and FO-PPR had a good linear relationship are in the concentration 2.5 ~ 50 μg / ml . And LC-MS detection limit of magnitude lower than the FO-PPR.(2) Using glycine buffer myoglobin is successfully desorbed from the sensor chip, but the data of recovery and reproducibility are not unsatisfactory. The OVA in the glycine buffer decompose quickly. Desorbed OVA is not able to determine quantitatively.