近年研究常使用兩性物質如:聚電解質和脂質作為穩定液晶液滴的物質。本實驗提出利用具有生物活性的寡肽，修飾在液晶液滴表面來穩定液晶液滴。藉由自訂序列的寡肽作為兩性分子修飾在液滴表面，製備出穩定液晶液滴。此液晶液滴經由蛋白酶對寡肽特異性的裂解，使液晶液滴表面排列由平行轉換至垂直，藉由液滴組態的改變來檢測蛋白酶，且這些液晶液滴能不聚集且不改變組態穩定30天，這顯示寡肽修飾的液滴具有生物檢測的潛力。此外本實驗針對寡肽序列對液晶液滴穩定性的影響做了研究。研究結果發現由親水性側鏈胺基酸lysine (K)與疏水性側鏈胺基酸leucine (L)、phenylalanine (F)和tyrosine (Y)組成的寡肽序列能穩定液晶液滴。
寡肽在不同生物檢測系統被廣泛作為識別分子偵測不同分析物。本實驗希望藉由多樣的寡肽序列來修飾液晶液滴，檢測各種目標。除了檢測應用外，液晶液滴的製備過程簡單且成本低。讀出系統的信號不需要複雜的儀器，結果可以容易被未經訓練的民眾解釋。這些優勢展現液晶液滴檢測系統具有極大的發展潛力。

Recent studies on the fabrication of liquid crystal (LC) droplets usually applied amphiphiles such as polyelectrolytes and lipids at the LC/aqueous interface to stabilize the droplets. In this work, LC droplets were prepared with bioactivity by decorating oligopeptide at the LC/aqueous interface for the first time. The LC droplets were fabricated by assembling a custom-made oligopeptide as amphiphiles at interfaces. These LC droplets can be used to detect proteases through the specific cleavage of protease on an oligopeptide leading to a bipolar-to-radial transition of LC configuration inside the droplets, and these LC droplets are quite stable as they can withstand coalescence without changing their configuration for 30 days, which demonstrates their potential in biological sensing applications. 
Besides, we also investigated the effect of oligopeptide sequence on the stability of LC droplets. It was found that the LC droplets are stable only when they are formed by oligopeptide composed of the amino acids with both hydrophilic side chain such as lysine (K) and hydrophobic side chain such as leucine (L), phenylalanine (F), tryptophan (W) and tyrosine (Y). Because oligopeptides are widely used as the probe in different biological detections, this information provides us the strategy for designing oligopeptide-decorated LC droplets to detect various targets. In addition to sensing applications, the fabrication of LC droplets is straightforward and cost-effective. The read-out of this system does not require sophisticated instrumentation, and the result can be easily interpreted by the untrained users. These advantages show the potential of LC droplets in commercial applications.

第一章 緒論	1
1-1	胜肽	1
1-2	胜肽感測器(Peptide-based sensors)	2
1-2-1 蛋白質胜肽感測器(Peptide-based protein sensor)	2
1-2-2 金屬胜肽感測器(Peptide-based metallic ion sensors)	3
1-2-3 蛋白酶胜肽感測器(Peptides-based enzymatic sensors)	4
1-3	液晶	6
1-3-1 液晶的形成	7
1-3-2 液晶的種類	8
1-3-3 液晶的排列	9
1-4	液晶型分析檢測	12
1-4-1 液晶型分析檢測作用機制	12
1-4-2 液晶型分析檢測介紹	14
1-4-2-1液晶-固體介面(LC/solid interfaces)	14
1-4-2-2液晶-水溶液介面(LC/aqueous interfaces)	17
1-4-2-3液晶液滴(LC droplets)	19
1-4-3 液晶檢測系統的優點	22
1-5	研究動機	23
1-6	總結	25
第二章 實驗儀器與方法	26
2-1 實驗藥品與材料	26
2-2 實驗儀器	27
2-3 實驗方法	29
2-3-1 溶液配製方法	29
2-3-2 製備塗覆DMOAP玻璃	34
2-3-3 空白玻璃製備	34
2-3-4 製備寡肽修飾的液晶液滴	35
2-3-5 液晶液滴的鑑定	35
2-3-6蛋白酶檢測	36
2-3-7 PCT臨界微胞濃度測試	36
2-3-8 利用HPLC分析裂解的PCT片段	37
第三章 結果與討論	38
3-1 玻璃基板的選定	38
3-2 LC droplets最佳條件建立	40
3-2-1 交聯劑(Glutaraldehyde, GA)濃度探討	41
3-2-2 還原劑(NaBH3CN)濃度探討	43
3-3 PCT濃度對LC droplets組態的影響	45
3-4 PT濃度對LC droplets組態的影響	47
3-5 不同濃度PCT droplets螢光圖	49
3-6 PCT droplets表面電位的探討	50
3-7 PCT濃度對LC droplets穩定度影響	51
3-8 PCT臨界微胞濃度測量	53
3-9 利用PCT droplets檢測蛋白酶	55
3-10 PCT droplets檢測蛋白酶螢光圖	57
3-11 HPLC分析裂解的PCT片段	58
3-12 PCT droplets偵測極限的探討	60
3-13 PCT droplets檢測特異性探討	62
3-14 不同序列oligopeptide對LC droplets穩定度的影響	65
3-15 pH對疏水胺基酸的影響	66
3-16 PKL對LC droplets穩定度的影響	67
3-17 PKF對LC droplets穩定度的影響	68
3-18 PKF與PKL對LC droplets組態的影響	69
第四章 結論	71
第五章 未來展望	74
參考資料	78

 
圖目錄
圖1 液晶特殊的光學訊號	6
圖2 液晶分子示意圖	7
圖3 可能的液晶分子結構	8
圖4 熱致型液晶結構圖	9
圖5 桿狀型液晶示意圖	11
圖6 圓盤狀液晶示意圖	11
圖7 液晶型分析檢測機制示意圖11	12
圖8 偏光顯微鏡交叉偏光板示意圖，(a)未含待測物質(b)含待測物質	13
圖9 DMOAP結構圖	16
圖10 液晶-固體介面示意圖，(a)基表面塗覆自組裝修飾分子，使液晶垂直排列(b)待測物與自組裝修基板作用，造成液晶分子擾動	16
圖11 (a) TEM銅網俯視圖(b)液晶-水溶液介面裝置側視圖(c)未受待測物影響液晶垂直排列及光學訊號(d)受待測物影響液晶垂直排列及光學訊號	18
圖12 液晶液滴常見的兩種組態圖(箭頭表示缺陷點)及示意圖(黑色線為液晶分子排列方向)	20
圖13 不同玻璃介面LC droplets的狀態	39
圖14 不同GA濃度製備的PCT droplets粒徑大小	42
圖15 不同還原劑濃度的 PCT droplets粒徑大小	44
圖16 PCT修飾於LC droplets表面示意圖，黑色實線表示液晶分子排列方向，彩色曲線表示PCT分子	46
圖17 不同濃度PCT droplets在塗覆DMOAP的玻璃表面上，於偏光顯微鏡下的LC droplets圖像	46
圖18 胺基酸分子結構	48
圖19不同濃度PT droplets在塗覆DMOAP的玻璃表面上，於偏光顯微鏡下的LC droplets圖像	48
圖20 標定Cy3螢光分子的PCT droplets	49
圖21利用比例尺量測修飾不同濃度PCT的 LC droplets粒徑大小	53
圖22 利用pyrene分子測定不同濃度的PCT溶液紫外光吸收強度，並將第三放光波長與第一放光波長訊號強度比，得到PCT的臨界微胞濃度	54
圖23 蛋白酶水解切割修飾於LC droplets的寡肽機制圖，實線表示液晶分子的排列方向，彩色曲線表示PCT分子	56
圖24 PCT droplets檢測不同蛋白酶，1000

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