- 學位論文
陣列式表面電漿共振於病毒檢測之最佳化開發
Optimization of Virus Detection Biosensor on Surface Plasmon Resonance Array
摘要
抗體陣列是作為蛋白質體學研究的一種新穎技術，可以高通量檢測多種蛋白質。由於抗體結構的特殊性，將抗體固定於感測表面已成為構建抗體陣列中的關鍵步驟。因此為了進行抗體固定化方法的最佳化，在本論文中，將以家禽白血病J亞群病毒為材料，用於病毒檢測之最佳化開發，以提供高效率、具有成本效益的陣列式表面電漿共振感測器。 在實驗中，為了建立家禽白血病J亞群病毒抗體最佳固定化的模式而提高感測器靈敏度的目的，因此比較了化學性和生物性固定抗體於金膜表面的方法。在固定了50μg/ml的蛋白質生物層上，檢測6.7-667 nM抗體的相對結合強度，由得到的資料可以計算出平衡常數和表面覆蓋率。此外，我們還評估了空間中立體效應和質量傳輸對於感測器靈敏度的影響。並且使用不同濃度家禽白血病J亞群病毒對167nM抗體的強度變化去建構一個濃度曲線圖，以期可以適用於陣列式表面電漿共振的檢測。
並列摘要
Antibody array is a novel technology that holds great promise in proteomics and diagnosis. It can also provide parallel analysis of multiple proteins in small sample volumes. Because of the high specificity of monoclonal antibody structure, it is thus crucial for the immobilization of antibodies to have correct orientation on the sensor surface. To optimize the immobilization of the antibody for small protein detection, the interaction between subgroup J avian leukosis virus and its monoclonal antibody is chosen as a model system in this thesis. In this work, we compared between the chemical and biological immobilization methods for the optimal conditions of the relative binding intensity in the range of 6.7-667 nM avian antibodies with 50μg/ml of the protein capturing layer on the gold surface. The measurement data from antibody-protein G and antibody-protein A interaction gives the equilibrium constants and the fractional surface coverage. In addition, the steric effect and mass transport under different flow rate are evaluated by a surface plasmon resonance biosensor. Using different concentration of subgroup J avian leukosis virus antigen to 167nM antiboby, we have constructed a concentration curve which can then apply to the surface plasmon resonance biosensor array for ALV detection.