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標題: 自我摻雜聚苯胺修飾電極結合奈米金粒子信號放大效益應用於電化學免疫感測器
gold nanoparticles modified self-doping polyaniline electrode for signal enhancement of electrochemical immunosensors
作者: 黃聖祐
Huang, Sheng-Yu
關鍵字: 競爭型免疫感測器
competitive immunosensor
self-doping polyaniline
human serum albumin
colloidal gold
出版社: 化學工程學系所
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摘要: 本研究利用自我摻雜聚苯胺 (self-doping Polyaniline) 於導電ITO玻璃 (indium tin oxide) ,製備一競爭式免疫感測器;當抗原與對應抗體結合時所產生之複合物,會造成阻抗值的變化,因應此效應即可使用差式脈衝伏安法 (differential pulse voltammograms) 量測對應於抗原濃度之電流變化,將之應用於人類血清白蛋白 (human serum albumin,HSA) 檢測上。 相較一般導電高分子組裝之免疫感測器,此研究利用摻雜2-胺基苯磺酸(o-aminobenzenesulfonic acid),使聚苯胺自摻雜形成圓球形態,達到高表面積目的,同時又不失去聚苯胺原本之高傳導性、高生物適性、高穩定性等優良性質,進而擴增感測濃度範圍及增強信號。 研究中使用競爭型的方式間接檢測抗原抗體的結合,其後在利用二次抗體 (goat anti-mouse) 結合膠體金作為放大信號的標的物,其中固定化抗原的濃度、競爭抗體的最高濃度及膠體金結合二抗的最佳濃度分別為250 μg/mL、20 μg/mL、5 μg/mL,最適化後檢測極限為1 μg/mL線性範圍可到10~200 μg/mL,添加膠體金粒子放大信號後可將檢測範圍降至0.5~200 ug/mL ,檢測極限為413 ng/mL 。 此競爭型免疫感測器在組裝上具有製備容易、低成本,在檢測上則有寬廣的線性範圍與良好的信號響應等等優點,並可適用單次檢測、用完即拋式的檢測器上,使用於臨床檢驗分析人類血清白蛋白HSA 。
A competitive immunosensor, based on the indium tin oxide(ITO) electrode modified by self-doping polyaniline(SPAN), was fabricated for the detection of human serum albumin(HSA) by measuring the differential pulse voltammograms(DPV). Compared to other immunosensors applying electrodes modified by conducting polymer, this study o-aminobenzenesulfonic acid(SAN) dopped Aniline(AN) to make the morphology of AN in the form of microsphere, resulting in the boost of signal, the extension of detection range, and the sustainability of high conductivity, high biocompatibility and high stability of AN due to high surface area of microsphere. In this study, a competitive binding of antigen in samples or on test line to antibody was adopted as the principle to indirectly determine the concentration of antigen in samples. In addition, secondary antibodies(goat anti-mouse) coupled with colloidal gold were used as the marker to amplify the signal of immunosensors. The concentration of the immobilized HSA, the highest concentration of anti-HSA and the concentration of secondary antibody conjugated with colloidal gold were found as follows: 250 μg/mL, 20 μg/mL, 5 μg/mL. The linear range was from 10 to 200 μg / mL with a optimal detection limit of 1μg/mL. After adding colloidal gold particles to amplify the signal, the linear range was from 0.5 to 200 μg/ml with a detection limit of 413 ng/ml. The proposed competitive immunosensor is easy to prepare, with the advantages of low-cost, wide linear detection range and good signal response. This assay provides a clinical testing method for measuring human serum albumin concentrations via single-use competitive immunosensor of ITO electrode modified by SPAN.
其他識別: U0005-3007201214492900
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