細胞膜蛋白感測晶片之研製 -使用氧化銦錫雙電極之連續阻抗量測

Taipei Medical University Institutional Repository > 醫學工程學院 > 生醫材料暨組織工程研究所 > 博碩士論文 > Item 987654321/4280

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題名:	細胞膜蛋白感測晶片之研製 -使用氧化銦錫雙電極之連續阻抗量測 Development of bio-chips for cellular-membrane-protein detection - continuous impedance measurement by ITO di-electrode
作者:	林君彥 Chun-Yen Lin
貢獻者:	口腔科學研究所
關鍵詞:	生物感測器氧化銦錫細胞膜蛋白細胞電阻抗量測 Biosensor indium-tin oxide ITO Cellular membrane protein Electric cell-substrate impedance sensing
日期:	2007
上傳時間:	2009-08-28 14:24:48 (UTC+8)
摘要:	細胞膜蛋白佔全部蛋白的30％，為蛋白質體學計畫中的研究重點。然而目前蛋白質相關的研究方法難以對細胞膜蛋白做非破壞的檢測，也無法長時間對單一樣本做連續變化的觀察。基於此，我們製作出以ITO為電極材料的生物晶片，非破壞性的連續測量細胞的交流阻抗。在本研究中MG-63細胞株培養於自製的ITO玻璃晶片上，當細胞成長數目越多時，電阻抗的變化量也越大，此與測量細胞活性之MTT法比較，有極高的線性相關(R2=0.9883，p<0.005)。本研究亦進一步結合免疫染色的原理，將帶有奈米級金粒子的抗體連結到細胞膜上Integrin β1蛋白，當有越多的Integrin膜蛋白存在時，細胞膜上被連結的金粒子也越多，此時會產生越大的電阻抗變化；與螢光染色法之讀值比較，亦有良好的線性相關(R2=0.9178，p<0.005)。本研究以較經濟的ITO晶片做為細胞膜蛋白非破壞性之研究方法，並為將來藥物篩選、癌細胞檢驗之應用性提供了初步的數據，但仍需要更多實驗佐證細胞膜蛋白量測相關之結果。 Membrane proteins, about 30% of the total proteins, become the major research topics of proteomic projects. There were many techniques were proposed for membrane proteins investigations. However, a method for continuous and long-term protein detection is still unavailable. Accordingly, in this study, we developed an ITO-based biochip for non-destructive membrane protein detection. In our study, when MG-63 cells were seeded on the ITO-based glass chips, a liner relationship was found when comparing with MTT assay (R2=0.9883, p<0.005). Additionally, when Integrin β1 membrane protein were taken into account, there exist a liner relationship between cell impedance and immuno-fluorescence intensity (R2=0.9178, p<0.005). In conclusion, we developed a novel ITO biochip for non-destructive membrane protein detection, and provided some preliminary results for future applications. It will be a useful tool for drugs discovery and cancer cells testing. The results obtained from this study can be a useful reference for future advance investigations on membrane-proteins detection.
資料類型:	thesis
顯示於類別:	[生醫材料暨組織工程研究所] 博碩士論文

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