侷限型表面電漿共振感測平台結合自動化微流道控制系統進行免標定及即時多重生物標記物檢測

細胞因子的檢測是判斷自體免疫的重要指標,除了解細胞行為的研究價值外,還可在臨床上提供許多診斷資訊,包括傳染病、癌症、自體免疫疾病、過敏原和藥物開發等資訊。然而標準的細胞因子檢測往往需要冗長的反應時間（8-24小時）及繁雜的標記步驟,因此常常會錯過治療的黃金時間 (數小時),且添加標定物進行檢測的方式僅能提供最終的細胞因子濃度,並無法即時檢測快速變化的免疫系統。為解決此問題本論文提出一利用侷限型表面電漿共振 (LSPR) 檢測細胞因子的平台。此平台由三個部分所組成：(1) 雙模式微流道晶片：根據不同需求切換至同步或獨立模式； (2) 自動化微流體系統：自動化控制試劑流速與體積並將試劑輸送至檢測區； (3) 光譜系統：執行免標定與即時生物分子檢測。本論文首先透過有限元素分析軟體模擬最佳化微流道的設計,接著驗證免疫量測的步驟、光譜系統、LSPR晶片以及自動化微流體系統。最後我們開發的LSPR平台可在4.5小時內完成自動化、免標定、即時和多通道平行化細胞因子檢測,此平台較傳統使用ELISA方式進行的檢測方式量測快2到6倍。我們相信此LSPR平台能克服傳統細胞因子量測的限制,藉由快速且免標定的生物分子檢測,來達成即時診斷與治療。

關鍵字

微流道；侷限型表面電漿共振；細胞因子

並列摘要

Cytokine detection allows the immune system to be monitored, providing useful information related to infectious diseases, cancer, autoimmune diseases, allergy transplantation, and drug discovery. However, conventional methods of cytokine measurement usually require multiple labor-intensive labeling procedures as well as long processing time (8-24 hours). Besides, the requirement of labeling process lead to only end-point readout measurements, which makes the monitoring of dynamic cytokine concentration variation impossible. Moreover, the long processing time hinders diagnosis and precludes timely treatments. In this thesis, a localized surface plasmon resonance (LSPR) platform is proposed to address these problems. The LSPR platform consists of three components: (1) a dual-mode microchannel which can switch into the synchronized or independent mode based on different assay steps; (2) an automatic microfluidic system which can control all reagent flow; and (3) a spectroscopy setup which performs label-free and real-time biomolecular detection. Our study first optimized the microchannel design through simulations. Then, the performance of the immunoassay protocol, the spectroscopy setup, the sensor, and automatic microfluidic system are validated in succession. Finally, an automated, label-free, real-time, and multi-parallel cytokine detection is performed by our LSPR platform in under 4.5 hours, which is 2-6 times faster than conventional methods. We believe our LSPR platform have the potential to overcome the limitations of conventional cytokine measurements and perform rapid cytokine detection for punctual diagnosis and timely treatments.