跨膜蛋白-細菌視紫質 (bacteriorhodopsin, BR)，被光照射後會吸收光能使質子從細胞膜膜內被推向膜外，因而產生跨膜質子濃度梯度，再透過外部電路連接即可產生光電流訊號。 BR乃來自古生嗜鹽菌Halobacterium salinarum的紫色細胞膜 (purple membrane, PM)；先前我們利用BR光電流響應和入射光強度間所呈現的正關係，已開發出PM光電感測晶片。本論文分為三部分，首先將可與三磷酸腺苷 (adenosine triphosphate, ATP) 結合的核酸適體 (ATP aptamer) 固定化於PM晶片上，再將與ATP aptamer部分互補並修飾有奈米金 (gold nanoparticle, AuNPs) 之去氧核酸序列 (complementary DNA, cDNA) 雜合上去，以製備出可檢測ATP之AuNPs-cDNA/ATP aptamer-PM複合晶片。研究以PM光電感測晶片所產生之光電流變化及拉曼光譜分析探討晶片製程及最適化檢測ATP條件。研究證實ATP存在時可與ATP aptamer結合並使AuNPs-cDNA脫附，而造成原本受AuNPs遮蔽之PM光電感測晶片的光電流回升，且此回升率與ATP濃度有關；在最適化條件下，30分鐘即可完成ATP檢測。當ATP濃度為200 μM時，PM光電流回升率可達87.64%，且最低檢測靈敏度可達0.1 nM ATP。其次為白血球 (leukocyte) 檢測晶片之開發，在PM晶片上依序固定化protein A與未被修飾之抗白血球抗體，即可利用白血球可遮光特性，檢測與定量白血球濃度。最後將4種可能與革蘭氏陽性菌 (gram positive) 細胞壁中之壁脂酸 (lipoteichoic acid, LTA) 結合之核酸適體 (LTA aptamer) 直接固定化於PM晶片表面以開發革蘭氏陽性菌檢測晶片，實驗結果發現其中2種LTA aptamer具有較高的微生物結合力，然而其特異性並不佳，有待進一步探討。

The transmembrane protein, bacteriorhodopsin (BR), absorbs light upon illumination and pumps protons from the cytoplasmic to the extracellular side of its cellular membrane, leading to a transmembrane proton gradient that can be used to generate photocurrents through an external circuit. BR is produced from the purple membrane (PM) of archaeon Halobacterium salinarum. We have previously developed PM-based photoelectric sensor chips based on the positive correlation between BR photocurrents and the incident light intensity. There are three parts in this thesis. In the first part, an ATP aptamer that specifically binds with adenosine triphosphate (ATP) was first immobilized on a PM chip. Complementary DNA (cDNA) partially complementary to the ATP aptamer and conjugated with gold nanoparticles (AuNPs) was hybridized on the ATP aptamer–PM chips to prepare AuNPs-cDNA/ATP aptamer-PM complex chips. The monitoring of the sensor chip production and the optimization of ATP detection conditions were investigated by Raman spectroscopy and photocurrent measurements, which was generated from the PM chips. The results evidenced that ATP binding led to the desorption of AuNPs-cDNA form the ATP aptamer–PM chip and thus caused the photocurrents of the chip to recover, which had been decreased due to the light-shielding effect of AuNPs, The photocurrent recovery level depended on ATP concentrations. ATP detection was accomplished in 30 min at the optimal condition. A 87.64% photocurrent recovery was obtained at 200 μM ATP, and a detection limit of 0.1 nM ATP was achieved. In the second part, leukocyte detection chips were developed by sequentially conjugating protein A and unmodified anti-leukocyte antibodies on PM chips. White blood cells blocked light significantly so that their concentrations were readily determined by the photocurrent reduction level. In the final part, four LTA aptamers with possible lipoteichoic acid (LTA) binding affinity, the major component in the cell wall of Gram-positive cells, were directly immobilized on PM chips to develop Gram-positive bacteria detection chips. The results showed that two of the four LTA aptamers had higher microbial binding abilities than the others, while their specificity needed to be further improved in future studies.

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