以陣列式QCM生物感測器偵測蛋白質濃度之研究

Measurement of Protein Concentration Using QCM-Array Biosensors

張啟霆(Chi-Tim Chang)

指導教授 ： 張耀仁

中原大學/工學院/機械工程學系/碩士(2007年)

https://doi.org/10.6840/cycu200700816

摘要

利用標準生化螢光反應，偵測特定蛋白質濃度，是目前在各類型檢測中最常使用的一種方式，其主要原因在於蛋白質接合專一性佳以及技術發展純熟，可一次性的檢測多種蛋白質並做重複性的實驗以確保實驗準確性，而其缺點在於需利用大量的抗體及螢光分子，並於每道接合程序皆需花費相當的時間以利雜交反應所需的時間，而在最終則需要以高階解析度掃描儀作為螢光分子亮度的辨識器，因此在金錢以及時間上的耗費相當可觀，並難以突破現今生物晶片所要求的即地(In-Situ)和即時(Real-Time)這兩個重要指標。
石英晶體微量天平(Quartz Crystal Microbalance)為利用質量負載(Mass-Loaded)原理，藉由生物辨識層的薄膜，可將蛋白質濃度準確辨識出，並再利用單一石英晶片將電極以多通道的方式製作於晶片上，即可達到高效率檢測，以及重複性檢測的目的，最重要在於可結合任一微流體及微陣列晶片，或直接將元件設計於石英晶片上，再利用共振電路以阻抗電訊號的方式擷取出量化後的數值。
本論文利用石英晶圓設計多通道電極，並利用硝化纖維薄膜(Nitrocellulose Membrane)及鎳(Nickel)金屬膜分別作為生物辨識層嘗試辨別蛋白質在不同濃度下所造成石英共振頻率的改變量，並利用螢光生化反應驗證其結果。實驗結果發現使用鎳金屬膜作為生物辨識層的共振頻率訊號較低，並不能有效分辨蛋白質濃度，而在螢光生化驗證的部份也未能產生訊號值。但若以硝化纖維薄膜作為生物辨識層批覆於電極上，則可以利用共振頻率準確分辨不同濃度蛋白質的變化，蛋白質濃度越高，共振頻率下降量越多，蛋白質濃度越低，共振頻率下降量越少，並在螢光生化檢測中，驗證其結果。

關鍵字

多通道石英晶體微量天秤 ； 螢光反應 ； 硝化纖維 ； 鎳

並列摘要

It is the common ways which detect specific protein concentration by using fluorescence of biologic reaction in all kind of detection. Because of the fluorescent particles have specific binding technique with protein molecule. It can do lots kind of different experiments at once and repeat it to make sure the accuracy. But it also has several faults. For example, it must be used plenty of antibodies and fluorescent particles and also take a lot of time on each of binding processes. Then, it needs a recognition component for fluorescence particles by a high-dpi scanner. So it still spends a lot of money for a machine. Actually, the most important thing is hard to break through the real-time and in-situ control by this way.
Quartz crystal microbalance which makes use of the mass-load theorem by biologic recognition component can recognize protein concentration accurately. Through the quartz wafer which fabricates multi-channel electrodes on can reach detection efficiently and repeatedly. It also may combine with micro-fluidic chip and micro-array chip.
In this paper, we designed multi-channel electrodes on one quartz wafer which covered with nitrocellulose membrane and Nickel to recognize different protein concentration by the quartz resonance frequency shift. We also use the fluorescence of biologic reaction to verify our experiment. In the experiment result, the resonance frequency which replace biologic recognition layer with nickel is too lower to differentiate protein concentration. It also didn’t have fluorescent signal on the biologic verification. Separately in the same way, we replace the biologic recognition layer with nitrocellulose membrane. It can recognize perfectly. The stronger protein concentration is, the lower resonance frequency is. On the contrary, the thinner protein concentration is, the higher resonance frequency is. We still use the fluorescence of biologic reaction to verify the experiment. The result is excellent.

並列關鍵字

nitrocellulose ； nickel ； multi-channel quartz crystal microbalance ； fluorescent reaction

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