長期以來，評估慢性糖尿病的控制狀態一直是一個難題，目前一般多以HbA1c(糖化血紅素)作為糖尿病人血糖監控指標，主因是紅血球的半衰期約為60天，測定紅血球之糖化血紅素可代表過去二至三個月的體內平均血糖值，且不受血糖量測誤差之限制。一般糖化血紅素偵測可利用Fructosyl Peptide Oxidase (FPO)氧化酵素，對結合於血紅素β鏈的N端胺基酸的血糖進行反應所產生的過氧化氫來做測量。因此本論文探討以兩種方式進行FPO酵素固定化，以為後續HbA1c檢測可達方便省時，操作簡易的目的。
第一種方式是固定化於電紡絲技術(Electrospinning)所製備的膜上，以濃度10% 聚乙烯醇溶液、施加電位14.5 kV-17.5 kV、針頭與屏幕距離125 mm 及流速6 μl/min 製備出無纖維球(bead)之聚乙烯醇電紡絲薄膜，並於酸性環境下將戊二醛與聚乙烯醇之羥基交聯，並利用膜上約7299.36 molecules/nm2之自由醛基與FPO上之胺基產生Schiff Base 反應而固定之 ; 第二種方式是固定化於四氧化三鐵磁性顆粒表面，先修飾上聚多巴胺，可測得表面胺基量為0.267 mmol/g-DW ，再同樣以戊二醛上醛基與聚多巴胺上胺基反應，亦可測得表面與FPO胺基反應之醛基含量為0.14 mmol/g-DW。
FPO固定於電紡絲薄膜後比活性為2.38 U/mg．mm2 ，在環境溫度 4-40 ℃ 、pH 值5-10 間皆有相當好之活性，此膜重複使用7次後能可有95 % 穩定活性，使用7天後約有60 %活性。FPO另固定於磁性顆粒上比活性可達0.465 U/mg．g-DW，在環境溫度 4-40 ℃ 、pH 值5-8有相當之活性，重複使用7次後可有70 %穩定活性，使用7天後約剩有50 %活性。由酵素動力學測試，比較未固定前FPO、固定於電紡絲薄膜之FPO及固定於磁性顆粒之FPO其三者之反應動力學參數Vmax/Km 值分別為1.18、0.002、0.06 min-1。

The measurement of glycated hemoglobin A1c (HbA1c) concentration in blood is generally regarded as a better method for assessing the effectiveness of diabetes treatment. The main reason is due to the fact that half-life of red blood cells is approximately 60 days, therefore measuring glycated hemoglobin of red blood cell may represent an average blood glucose levels for past two to three months. Generally, Fructosyl peptide oxidase (FPO) can react the sugar coupled on the N-terminal amino acid of β-chain to generate hydrogen peroxide and it can subsequently react with a dye via an enzymatic reaction to develop color so that HbA1c concentration can be determined. In this thesis, we develop two methods to immobilize FPO enzyme in a convenience, simple, and economic ways.
The first one is to electrospin to form a film without beads by 10 % (W/V) polyvinyl alcohol to form nanofibrous mat cross-linked with glutaraldehyde . The excess amount of free aldehyde group on the nanofiber was employed for FPO immobilization via surface Schiff Base reaction. The second one is to modify magnetic iron oxide particle surface with poly-dopamine followed by glutaraldehyde activation. The surface of magnetic particles activated by GA was employed for FPO immobilization.
The specific activity of FPO immobilized electrospun nanofibrous mat is 2.38 U / mg• mm2. The mat retained 95% of its initial activity after seven cycles of reuse, and 60% activity after 7 days incubation at 4 ℃. The specific activity of magnetic nanoparticles is 0.465 U / mg. g-DW, and also showed good activity in 4-40 ℃and pH 5-8. The magnetic patticles retained 70% its initial activity after seven cycles of reuse, and 50% activity after 7 days storage.

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