糖化血紅素的濃度為監控糖尿患者血糖值的一個重要指標，紅血球中糖化血紅素( HbA1C)所含比例反映病患近二~三個月內的平均血糖控制情況。本研究是利用硼酸分子可在微鹼性環境與醣類結構上之雙醇基團(cis-diol)產生親和性交互作用，進而分離人體血液中的糖化血紅素。首先以CM-瓊脂糖凝膠(CM-Sepharose)表面修飾上硼酸親合配體來測試吸附醣蛋白-HRP的能力，此自製之含有硼酸配體之瓊脂糖凝膠對於醣蛋白的回收率皆可達到90%以上，對HRP的親合吸附能力為17.93 mg HRP/g-dry Resin。而利用電漿表面共振儀可觀測到糖化血紅素分子與表面固定有硼酸配體之感測晶片間之交互作用，利用SPR共振角度位移量在365.9-486.5 mo之間可偵測到血液中糖化血紅素濃度在6.25-12.5%之間。此外，本研究應用表面帶有羧酸官能基之尼龍6,6薄膜(Negatively-charged Nylon 6,6)為固體基材，並藉由活化劑 1-(3-二甲氨基丙基)-3-乙基碳二亞胺鹽酸((N-(3-dimethylaminopropyl) -N′-ethylcarbodiimide hydrochloride , EDC)使3-胺基苯硼酸 (3-aminophenylboronate acid, 3-PBA)以共價鍵結合的方式固定在膜上，即成為一具有硼酸配體親和性薄膜，此親和性膜可來分離血液中糖化血紅素(Glycosylated Hemoglobin , HbAlC)。此分離出來之糖化血紅素仍具有如血紅素一般之過氧化酶活性，結合過氧化酶活性呈色法可檢測出糖化血紅素含量，本論文進一步探討利用濾紙取代比色槽作為基底，使糖化血紅速之過氧化酶活性於濾紙上所呈現呈色反應之顏色變化，以肉眼即可觀察並判斷血液中之血糖值高低，進一步可透過影像處理來量化紙上所呈現之顏色而達到定量檢測糖化血紅素的目的，結果顯示此方法可以偵測到血液中5.1-12.5%的HbAlc濃度。

Glycosylated hemoglobin (HbA1c) concentration is currently an important diagnostic indicator for the diabetic patient. The HbA1c concentration in blood reflects the average glucose concentration of the past 2-3 months. Boronic acids is known to form reversible covalent bonds with 1, 2- or 1, 3-diols to generate five-or six-membered cyclic esters in alkaline condition that can be used as an affinity ligand for specific binding the glycosylation sites of the HbA1c. CM-Sepharose was immobilized 3-aminophenylboronic acid as an affinity ligand on the surface for efficient enrichment of glycoproteins. Horseradish peroxidase (HRP) was used as a model glycoprotein to test the affinity adsorption efficiency of the boronic acid- CM-Sepharose. The percentage of HRP recovery was 90% and the adsorption capacity of HRP was 17.93 mg HRP/g-dry Resin. Real-time detection of immobilized 3-APBA on SPR chip interaction with HbA1C by Surface plasmon resonance (SPR) was also carried out. The results showes SPR angle shifts for binding HbA1c in the concentration of 6.25%-12.5% were 365.9 moand486.5 mo, is proportional to the HbA1C concentration. Since hemoglobin also has the function of peroxidase to catalyze the reduction of hydrogen peroxide with the oxidation of N-(Carboxymethylaminocarbonyl)-4,4'-bis(dimethylamino) diphenylamine sodium salt (DA-64) to generate color, in this study, phenylboronic acid was immobilized on porous Nylon membrane to specifically bind with HbA1c. The glycated hemoglobin eluted from the phenylboronic acid-immobilized membrane still maintains its hemoglobin peroxidase activity. The HbA1c was applied on the filter paper to develop green color by using dye DA-64 mixed with hydrogen peroxide. The intensity of color developed can be distinguished by visual recognition and digitalized by imageJ software that 5.1-12.5% HbA1C in blood was succefully determined. 

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