近年來由於基因微陣列技術的進步，人類的基因密碼逐漸被專家所解密，然而真正影響細胞活動的是蛋白質，癌細胞所產生的蛋白質是癌症篩檢中重要的判讀依據，因此可供檢測蛋白質微陣列之製作益形重要。本論文利用生醫微機電製程技術，製作批次且廉價之蛋白質晶片，使用的檢體量少且可拋棄式的優點可避免檢體交互汙染，人性化操作介面可使蛋白質失去活性前能即時快速地轉印到檢測晶片表面。晶片以厚膜光阻SU-8作微流道，底部則以高分子軟性矽膠材料作為微印章，蛋白質檢體以滴管由儲存槽注入，經過微流道到達底部微印章陣列，以印章蓋印方式製作蛋白質微陣列，整個過程中利用毛細力驅動不需額外施力。本研究製作各式蛋白質微陣列，探討並歸類造成微陣列缺陷成因。目前已成功製作出三種不同形式蛋白質微陣列而不造成交互汙染，包括單種蛋白質陣列、多種蛋白質陣列及癌症檢體陣列。主要使用之蛋白質檢體為標定Cy3螢光的兔子免疫球蛋白抗體、標定Cy5螢光的老鼠免疫球蛋白抗體、子宮頸癌抗原E6及肝癌抗原HuRP。微陣列其尺寸標準差於3%以內，螢光強度在20%以內。利用晶片所製作出之癌症抗原微陣列，經由傳統酵素免疫螢光分析法，得到癌症檢測最小解析度約為1ng/μl。

The protein chip is a micro-chip with its surface modified to detect various disease causing proteins simultaneously in order to help find a cure for them. This thesis is going to introduce a back-filling protein chip without any external driving force. Bio-MEMS technique makes a batch of cheap chips, and they are disposable form samples cross contamination. The flow channels are made by SU-8；PDMS polymer was used for the array at the bottom. First of all, the protein samples were lead into the upper reservoirs by a pipette. Next the samples flowed through the micro-channels by the capillary force and arrived the array at the bottom. Finally different protein arrays would come out like the term of printing stamps. This research improves the quality of arrays and summarizes the problems of the faults of the arrays. The used protein solutions are Cy3-tagged anti-rabbit IgG and Cy5-tagged anti-mouse IgG. Now this chip can successfully demonstrate three types of arrays without cross-talk, including single-protein array, multi-protein arrays and cancer-marker arrays. Each spot is formed by a single droplet about 1nl at each position. These protein arrays is characterized with a deviation ±3% in size and 20% in fluorescent intensity. The cancer antigens were made by chip stamping and using ELISA (Enzyme Linked- Immuno -Sorbent Assay) method the cancer levels down to 1ng/μl were detected.

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