分子印刷高分子是藉由模板分子與功能性單體所構成具有專一性辨識的人工抗體。由人類血清白蛋白中挑選出三種構型為無規則的線圈或螺旋結構的胜肽序列為模板分子，然後在經化學法修飾後的纖維素紙上，使用丙烯酸、丙烯醯胺、苯基丙烯醯胺與亞乙機雙丙烯醯胺做為官能性單體與交聯單體，經共聚合反應形成為披覆分子印刷高分子之纖維素紙。
由觀察顯示人類血清白蛋白溶液的螢光強度與其濃度成線性關係，因此利用螢光光譜儀，監測由印刷之纖維素紙吸附過的人類血清白蛋白溶液。並且將兩片已印刷之纖維素紙結合，形成具有雙面印刷之纖維素紙來提升吸附效果。
利用高靈敏度的石英晶體微天秤，來驗証、確認其印刷之晶片能專一性結合白蛋白。當以人類血清白蛋白568-582(螺旋) 作為模板分子時其印刷之晶片於吸附胜肽之Kd 值為 7 nM；而於吸附人類血清白蛋白時，其吸附值可以到達Kd為50 pM。

Molecularly imprinted polymers (MIPs) were fabricated with template and functional monomers to form artificial antibody with specific recognition ability. Three peptide sequences with random coil or α-helix configuration from human serum albumin (HSA) were selected as the template. The cellulose paper was chemically-modified and copolymerized with acrylamide, acrylic acid, N-benzylacrylamide, N,N’-ethylenebisacrylamide to form MIPs coated cellulose paper.
A linear relationship was observed between the fluorescence intensity to the concentration of human serum albumin. Therefore, fluorescence spectroscopy was applied to monitor the HSA solution after adsorption by MIPs cellulose paper. Moreover, two piece of MIPs cellulose paper were combined together as double-side MIPs cellulose paper to enhance the adsorption effect.
High-sensitivity quartz crystal microbalance is also applied to verify as well as to confirm the specificity of the MIPs on chip for binding with HSA. Upon using HSA568-582(α-Helix) as the template, the Kd values of the QCM chip was 7 nM for peptide adsorption. As for binding with HSA, the best Kd value was 50 pM.

總目錄 I
圖目錄 XIII
表目錄 IX
對照表 XIV

第一章緒論 1
1.1前言 1
1.2實驗設計 2
1.2.1MIFC運用於辨識HSA示意圖。 3
第二章、文獻探討 5
2.1人類血清白蛋白(Human serum albumin) 5
2.1.1前言 5
2.1.2人類血清白蛋白之結構 6
2.1.3人類血清白蛋白於臨床上之意義 8
2.2分子印刷高分子( molecularly imprinted polymers) 10
2.2.1前言 10
2.2.2原理 11
2.2.3分子印刷術之應用 14
2.3螢光光譜儀 (Fluorescence Spectrometer ) 16
2.3.1螢光的發現 16
第三章、研究內容 19
3.1藥品 19
3.2儀器設備 21
3.3聚合單體 22
3.4 Human serum albumin之抗原決定部位胜肽的選擇 23
3.5微波胜肽合成儀合成胜肽 26
3.5.1製備胜肽序列 LPRLVRPEVDV 26
3.5.2製備胜肽序列KEFNAETFTFHA 29
3.5.3製備胜肽序列LPSLAADFVESK 32
3.6.1 利用溶劑系統(H2O:CH3CN = 1:1)測試 38
3.6.2利用溶劑系統(H2O/CH3CN = 1/1)與(TFE / H2O = 7 / 3)測試 39
3.7分子印刷高分子之基材製備 40
3.7.1 利用氯化亞碸進行濾紙的表面修飾 40
3.7.2利用苄胺進行濾紙的表面修飾 42
3.7.3利用丙烯醯氯進行濾紙的表面修飾 44
第四章 結果與討論 47
4.1分子印刷高分子的親和力公式推導 47
4.2 HSA螢光偵測的範圍 50
4.2.1螢光光譜儀參數設定 50
4.2.2利用螢光偵測HSA標準樣品之檢量線 51
4.3製備具雙面吸附能力之高分子印刷濾紙 52
4.3.1單體的選用 52
4.3.2測試聚合溶液單體比例(AC:AM:BZ:EBAA=2:4:1:15) 52
4.3.3製作具雙面印刷之纖維素紙 52
4.4 HSA抗原決定部位印刷濾紙親合能力測試 53
4.4.1測試Rondom coil structure of double-side MIFC112-122對HSA的親和能力 53
4.4.2測試Rondom coil structure of double-side MIFC302-313對HSA的親和能力 54
4.4.3測試Rondom coil structure of double-side MIFC500-511對HSA的親和能力 55
4.4.4測試Rondom coil structure of double-side MIFC568-582對HSA的親和能力 56
4.4.5測試α-Helix structure of double-side MIFC568-582對HSA的親和能力 57
4.5測試聚合溶液單體比例(AC:AM:BZ:EBAA=1:1:2:4) 58
4.5.1製作雙面MIFC 58
4.5.2測試Rondom coil structure of double-side MIFC112-122對HSA的親和能力 59
4.5.3 測試Rondom coil structure of double-side MIFC302-313對HSA的親和能力 60
4.5.4測試Rondom coil structure of double-side MIFC500-511對HSA的親和能力 61
4.5.5測試Rondom coil structure of double-side MIFC568-582對HSA的親和能力 62
4.5.6測試α-Helix structure of double-side MIFC568-582對HSA親和能力 63
4.6數據分析 64
4.6.1以聚合單體比例(AC:AM:BZ:EBAA=2:4:1:15)測試各別吸附力 64
4.7利用聚合溶液比例 AC:AM:BM:EBAA=1:1:2:4的MIFC568-582對不同Protein 測試其專一性 74
4.8利用聚合溶液比例 AC:AM:BM:EBAA=1:1:2:4的MIFC來測試其再現性 75
4.9 HSA抗原決定部位印刷晶片親合能力測試 77
4.9.1 HSA112-122之印刷晶片對HSA112-122親和能力之測試 77
4.9.2 HSA500-511之印刷晶片對HSA500-511親和能力之測試 79
4.9.3 HSA302-313之印刷晶片對HSA302-313親和能力之測試 81
4.9.4 HSA568-582之印刷晶片對HSA568-582親和能力之測試 83
4.9.5 HSA568-582之印刷晶片對HSA568-582親和能力之測試 85
4.10利用印刷之晶片對HSA是否具吸附能力 89
4.10.1測試HSA112-122印刷之晶片對HSA的吸附能力 89
4.10.2測試 HSA302-313印刷之晶片對HSA的吸附能力 91
4.10.4 Rondom coil structure of HSA568-582印刷晶片對HSA親合能力測試 95
4.10.5 α-Helix structure of HSA568-582印刷晶片對HSA親和力測試 97
4.11數據分析 99
第五章、結論 105
第六章、參考文獻 107

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