氧化石墨烯（graphene oxide, GO）具獨特的材料特性，為近年來的研究焦點。紫膜（purple membrane, PM）為Halobacterium salinarum嗜鹽古菌的細胞膜，其內的細菌視紫質（bacteriorhodopsin）蛋白具有光驅動單方向質子泵功能。本研究將GO應用於PM生物晶片技術上，並以水滴接觸角、微分光電流、脈衝雷射光電流、傅立葉轉換紅外光譜、蛋白質定量、螢光分析等分析進行探討其影響。先在經過自排性單分子膜胺基化的ITO玻璃上塗覆活化avidin或與GO之混合物，再以生物性親和吸附作用將經biotin修飾的PM（b-PM）固定化。水滴接觸角會從清潔後的ITO表面5 - 8 °增加至自排性單分子膜的約30 °，再塗覆上活化avidin或與GO之混合物後上升到約65 °，但隨著GO添加濃度的增加，接觸角會緩緩下降至15 °左右，顯示GO可增加晶片表面的親水性；成功塗覆b-PM後的水滴接觸角則維持在30 °左右。GO的微量添加對晶片的微分光電流影響不大，但GO濃度增加至臨界濃度時，光電流則幾乎完全喪失，僅剩餘約5 %。以脈衝雷射激發PM晶片，發現GO存在時，B1訊號強度及成功被激發的機率提高許多，意謂PM的定向性提高。傅立葉轉換紅外光譜分析發現活化avidin與GO混合後，氫鍵及C-N拉伸訊號的強度均增加，推測氫鍵及C-N鍵結是活化avidin與GO結合的主因。BCA蛋白質定量分析透析過後的活化avidin及與GO之混合物，發現添加GO可使活化avidin的保留率提升20 %。最後螢光分析結果顯示，活化avidin的最大放射波長不因GO的添加而紅位移，立體結構不受影響。總結本研究發現，GO可與活化avidin結合，作為以生物親合固定化法製作GO-PM複合晶片的架橋，此外，適量的添加GO並不會影響晶片活性，且有助於PM貼附的定向性。

Graphene oxide (GO) has unique properties and has recently become a research focus. Purple membrane (PM) is the cellular membrane of archaeon Halobacterium salinarum, which contains bacteriorhodopsin with a unidirectional light-driven proton pump ability. This research applied GO to PM-chip technology and employed water contact angle, CW differential photocurrent, pulse-laser fast photocurrent, Fourier transform infrared spectroscopy (FTIR), protein quantification, and fluorescence analyses to examine the GO effects. Self-assembled-monolayer (SAM) fabricated ITO was first coated with either activated avidin or its mixture with GO and used as linkers to immobilize biotinylated PM (b-PM) through affinity adsorption. The contact angle of clean ITO was 5~8, rose to ~30 after SAM fabrication, and further increased to ~65 following the coating with either activated avidin or its mixture with GO. The contact angle gradually decreased to ~15 with the increased addition of GO and was maintained at ~30 if b-PM had been successfully captured. A minor addition of GO caused insignificant effects on the differential photocurrent response of the as-prepared PM chips, while the response was almost completely inhibited with the result of only 5% residual activity when the GO addition concentration was increased and reached a critical value. Upon pulse-laser illumination, the chips prepared with the addition of GO generated a higher B1 fast photocurrent response with a better success rate than the chips prepared without GO, implying an improvement of the orientation uniformity of immobilized b-PM. FTIR analysis of the mixture of activated avidin and GO revealed augmentation of hydrogen binding as well as C-N stretching, suggesting hydrogen and covalent linkages between them. BCA protein analysis showed that 20% more activated avidin could be retained in dialysis tubes if GO had been added. Furthermore, there is no red shift of the fluorescence spectra of activated avidin when GO was present. Therefore, this study suggested the functional linkages between activated avidin and GO and demonstrated the feasibility of applying their mixture as a linker for b-PM affinity immobilization with little effects on PM activity but improvement of PM orientation uniformity.

陳逸航，"定向性細菌視紫質晶片之光電與二倍頻響應探討"，國立臺灣科技大學化學工程研究所碩士論文 (2010)

蔡孟訓，"自排性單分子膜於bacteriorhodopsin光電晶片之製備探討"，國立臺灣科技大學化學工程研究所碩士論文 (2011)

許遠玲，"氧化石墨烯於紫膜晶片技術之應用"，國立臺灣科技大學化學工程研究所碩士論文 (2012)

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