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標題: | 修飾低吸附雙極性多胜肽金表面於胎盤生長因子檢測應用 Surface Functionalization of Gold Surfaces with Polypeptide: A Low-Fouling Zwitterionic Surface for Detect Placenta Growth Factor |
作者: | Wei-En Hsu 許偉恩 |
指導教授: | 林啟萬 |
關鍵字: | 表面電漿子共振,抗吸附,兩性離子,大腸直腸癌,胎盤生長因子, Surface Plasmon Resonance,Anti-fouling,Zwitterion,Colorectal Cancer,Placental Growth Factor, |
出版年 : | 2017 |
學位: | 碩士 |
摘要: | 在台灣大腸直腸癌的發生已經率逐年提高,目前大腸直腸癌是全台灣所有癌症中發生率第一名的癌症,已經七度蟬聯癌症發生人數的第一名,且大腸直腸癌在死亡率上也高居第三,僅次於肝癌和肺癌的死亡率。根據過去的許多癌症研究中發現,目前對於大腸直腸癌的初期診斷,胎盤生長因子(PlGF)是一個重要的指標,可以在患病初期及早發現並治療,並且胎盤生長因子也可以當成預後復發的危險因子指標之一。
本研究為了能夠檢測胎盤生長因子的存在,以表面電漿子共振技術來進行量測,該技術具有高靈敏度、即時量測且不須螢光標記等優點,並且使用抗體為感測元件,能夠對於標的物有良好的結合能力。但由於胎盤生長因子是存在於血清中,血清檢體包含了大量的蛋白質,在接觸量測表面時會受到極大的非特異性吸附影響,導致錯誤的訊號產生。為了解決這個現象,本研究使用混合的胺基酸序列來進行蛋白質的抗吸附處理與抗體的固定化,以簡單的方式達到保護量測表面的同時,也可以用於檢測胎盤生長因子的存在。在溶液中,本研究所提出的量測方法已經可以有效的檢測出PlGF的存在,且檢測極限可以到達2pg/ml。在實際臨床樣本的量測中,由於非特異性吸附的影響,目前與現有ELISA技術相比,同樣可以擁有六成的精確度。 In Taiwan, the incidence of colorectal cancer has been increasing year by year. Colorectal cancer is the first occurrence of all cancer. The incidence rate of colorectal cancer has become first for seven consecutive years and the mortality rate also ranked third, only behind liver cancer and lung cancer. According to many cancer studies in the past, placental growth factor(PlGF) is the important marker that can help us to know the potential patient and give them some treatment. In addition to this, PlGF can also be used as one of the risk factors for prognosis recurrence. In this study, I want to know if PlGF exists in patient's serum. I used surface plasmon resonance(SPR) technology for detection of PlGF. The reasons being that SPR has many advantages, like high sensitivity, real-time detection, non-Fluorescent label and so on. Moreover, I used the antibody as my sensing element. It has a good ability to combine with the target. However, PlGF exists in the serum contains a large amount of protein and will be affected by non-specific adsorption, therefore resulted in the error signal. In order to solve this problem, I used the mixed amino acid sequence for protein anti-fouling and antibody modification in this study. This simple method can achieve protection of surface, and also can be used to detect if PlGF exists at the same time. In the solution, the measurement method proposed in this study has been able to effectively detect the presence of PlGF, and the limit of detection(LOD) is 2pg/ml. Due to the impact of non-specific adsorption, if compare the actual clinical sample with the existing ELISA technology, this study also have sixty precent accuracy. |
URI: | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/59432 |
DOI: | 10.6342/NTU201701042 |
全文授權: | 有償授權 |
顯示於系所單位: | 醫學工程學研究所 |
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