結合自由流電泳與表面增強拉曼光譜來進行多重分子檢測

Abstract

電泳是一種在各個領域被廣泛應用的樣品前處理技術。自由流電泳（FFE）是其中一種常見的電泳模式，因能連續且快速的進行分離，被用於細菌、蛋白質或DNA的樣品純化和濃縮。近年來，由於降低人為錯誤和提高檢測準確性的潛力，自由流電泳與不同的偵測技術如電化學或是質譜儀的整合變得愈發受歡迎。然而，現有的整合方法面臨諸多限制，包括數據提供資訊不足、檢測時間較長和高成本等。為了應對這些挑戰，表面增強拉曼散射（SERS）因其高靈敏度和高特異性而備受矚目。在本篇論文中，我們開發了一個無縫整合FFE和SERS的多重分子檢測平台，可以在二十分鐘內完成樣本前處理與訊號檢測。我們首先通過使用不同電特性的三種螢光分子來評估平台的性能，驗證了此平台能夠成功分離並同時偵測與SERS擁有不同親和力的分子。同時我們也透過模擬與實驗的交互驗證，建立了最佳的操作參數來進行分離。並且依據這樣的模擬公式，也能分析樣本的電特性，藉此獲得更多資訊。最後我們通過分離與檢測嘌呤的混合樣本證明了此平台能用於細菌分析，同時還能透過調整環境pH值來進行更廣泛的應用。FFE與SERS的整合為同時檢測多種分子提供了一個具備了高靈敏度以及更快的處理速度的方法，有潛力應用於臨床診斷、環境監測和食品安全。

Electrophoresis is a widely used sample pre-processing technique in various fields. Free Flow Electrophoresis (FFE) is a common electrophoresis mode known for its continuous and rapid separation capabilities. It is applied in the purification and concentration of samples such as bacteria, proteins, or DNA. In recent years, integrating FFE with different detection techniques, such as electrochemistry or mass spectrometry, has gained popularity due to its potential to reduce human errors and enhance detection accuracy. However, existing integration methods face various limitations, including insufficient data information, prolonged detection times, and high costs. To address these challenges, Surface-Enhanced Raman Scattering (SERS) has garnered attention for its high sensitivity and specificity. In this thesis, we developed a seamlessly integrated platform that combines FFE and SERS for the detection of multiple molecules. This platform allows for sample pre-processing and signal detection to be completed within twenty minutes. We first evaluated the performance of the platform by using three fluorescent molecules with different electrical properties, confirming its ability to successfully separate and simultaneously detect molecules with varying affinities for SERS. Additionally, through the validation of simulations and experiments, we established optimal operational parameters for the separation process. By utilizing simulation formulas, we could analyze the electrical characteristics of the samples, providing more comprehensive information. Finally, we demonstrated the versatility of the platform by separating and detecting a mixed sample of purine derivatives, showcasing its potential for bacterial analysis. The platform's adaptability for broader applications was further highlighted by adjusting the pH value.