古生嗜鹽菌Halobacterium salinarum 之紫色細胞膜，簡稱紫膜 (purple
membrane, PM) ，含有細菌視紫質 (bacteriorhodopsin, BR) ，受光激發後會產生
光循環並將質子從細胞膜內側傳遞到細胞膜外側，形成一個光驅動質子泵
(light-driven proton pump)，再外加電極與導線即可用以產生光電流訊號。我們已
利用BR 光電流響應與入射光強度呈正向關係之原理，製備出一系列PM 微生物
光電晶片。本研究共分為兩部分，第一部分先將一組互補的脫氧核酸 (DNAzyme)
與鍵結有奈米金 (gold nanoparticle, AuNPs) 之基質寡核酸 (substrate oligo) 依序
塗覆至PM 晶片上，以作為生物辨識分子，並利用AuNPs 可遮光之事實來檢測
Pb2+。研究中探討晶片製程及最適化檢測條件，並且以PM 感測晶片之光電流與
拉曼光譜分析，分別確認Pb2+檢測條件及製程可行性。研究發現，PM 感測晶片
光電流回升程度與Pb2+濃度有關，當Pb2+達1 μM時光電流可有最大回升比例 (97
%) ；此Pb2+感測晶片具有高選擇性，在最適化條件下，15 分鐘即可完成Pb2+
檢測，且晶片檢測靈敏度可低達1 nM (0.208 μg/mL) Pb2+，遠小於法規所規定的
10 μg/mL。第二部分則是延續實驗室先前開發成果，深入探討可分別檢測血紅素
(hemoglobin, Hb) 與糖化血紅素 (glycated hemoglobin, HbA1c) 之Hb aptamer-PM
及HbA1c aptamer -PM 複合晶片的最適化aptamer 固定化濃度、Hb 與HbA1c 檢測
時間以及晶片保存性研究。在調整晶片製程至最適化後，兩者晶片檢量線的靈敏
度均較原先晶片製程為高；同時發現利用適合添加物，可將Hb 與HbA1c 檢測時
間由原先的2 小時縮短至15 分鐘。最後發現在最適化條件下保存一個月後，PM
與Hb aptmaer-PM 複合晶片均仍保有原有活性並可進行檢測，顯示其未來商業化
之可行性。

Purple membrane (PM) is one of the cellular membranes of archaeon
Halobacterium salinarum, containing bacteriorhodopsin (BR), a light-driven proton
pump capable of transporting a proton across PM upon illumination and subsequently generating photocurrents. Based on the linear dependence of BR photocurrents on illumination intensities, we had developed series of PM-based biosensor. This study contains two parts. First, based on light scattering of gold nanoparticle (AuNPs), a PM chip sequentially coated with DNAzyme and AuNPs-conjugated substrate oligo was used as the recognition element to detect lead ion (Pb2+). Chip fabrication and Pb2+ detection were both optimally investigated and confirmed by both Raman spectroscopy and PM-photocurrent measurement. The recovery of the photocurrent density linearly depended on Pb2+ concentrations, with the highest photocurrent density recovery (97 %) observed at 1 μM Pb2+ of the PM-based sensor chip. The Pb2+ sensor chip had a good selectivity and completed the detection in 15 min, with a detection limit of 1 nM (0.208 μg/mL), which was much lower than the regulation concentration (10 μg/mL). Secondly, we continued our previous hemoglobin (Hb) and glycated hemoglobin (HbA1c) studies with the developed Hb aptamer-PM and HbA1c aptamer-PM chips. By optimizing the fabrication process, the sensitives of both chips increased. In addition, the use of a proper additive speeded up detection, shorten the detection from 2 hr to 15 min. Finally, the stability studies revealed both PM and Hb aptamer-PM chips maintained their activities for at least one month, suggesting their
potentials in future commercial applications.

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