- 學位論文
以田口式實驗方法設計及製作最佳化鎳沉積之蛋白質晶片
Optimal Design of Nickel-Coated Protein Chips Using Taguchi Approach
摘要
在傳統的金屬親合層析法（Immobilized Metal Affinity Chromatography, IMAC）中經常利用鎳離子(Ni2+)來修飾層析管柱(chromatographic column)中beads表面，以利吸附histidines(his) -tagged 蛋白質而組成快速純化的分離系統，本實驗根據上述分離技術之方法做為實驗依據基礎，利用電鍍技術將鎳沉積於晶片基材表面，並以免疫學分析技術驗證鎳晶片也具有吸附之效果。在檢測方面利用高靈敏度之共厄焦式掃描器檢測螢光影像。本實驗以田口實驗規劃與配合直交表運用設計蛋白質晶片沉積，直交表設計法則規劃L18之實驗，達到最佳化實驗晶片之結果。本研究最重要之目的是將此層析技術使其平面化，運用做為蛋白質晶片吸附生物分子之關鍵技術，並成功地將蛋白質His-tag固定於晶片表面，發展新的固定蛋白質晶片，且有不錯的成效，提高再現性，並且降低晶片之成本。
並列摘要
For the traditional immobilized metal affinity chromatography (IMAC), the nickel is used to modify the surface of the beads in chromatographic column for capturing the histidines (his)-tagged protein to make up a rapid purification system. Based on IMAC, a nickel coated protein chip was developed by electroplating technology and its capture property was tested by the immunoassay with a high sensitivity of fluorescence confocal scanner. Taguchi method was adopted to determine the optimal process of protein chip. Hence, a series of L18 experiments by Orthogonal Array rule were performed. The major purpose of our research is the planarization of IMAC technology so that bio-particles can be absorbed on protein chip for further analysis. The advantages of this novel protein chip are good performance, high repeatability, and less cost.