HbA1c指人體血液中的紅血球所含之糖化血紅素，當血液中的葡萄糖進入紅血球，但和血紅素結合後就形成糖化血紅素。一般紅血球平均壽命約為120天，因此血中糖化血紅素的濃度可以反映體內最近2-3個月的血糖控制情況。分析HbA1c含量的方法有許多種，本論文是以FPO(fructosyl peptide oxidase)氧化經Neutral protease水解HbA1c所產生的fructosyl valyl histidine(FVH)產生H2O2，再利用酵素呈色法進行偵測，推算HbA1c的濃度。
本研究建構一套能夠利用酵素呈色方式測量血樣中HbA1c含量的程序，在60℃下中性蛋白水解酶最能有效水解血紅素，在活性單位(40KU/mL)水解酶下反應1.5小時就能將HbA1c水解釋出所有FVH片段，可以FPO進行氧化反應，所產生的H2O2可用HRP呈色反應偵測到5.2~15%的HbA1c，為能重覆使用FPO，而將FPO固定化於經聚多巴胺表面改質的PE膜上，發現經GA(glutaraldehyde)進一步修飾之聚多巴胺表面能提升FPO酵素的固定化量，此固定化的FPO在溫度4~60℃、pH 3~11下仍能保有相當的活性。固定化之FPO-PE膜可用於偵測FV濃度其線性範圍為0.02~0.7 mM，且使用至少7次擁有95%以上的活性，可用於測定血樣中之HbA1c濃度。

Glycated hemoglobin (HbA1c) concentration of blood is usually measured to identify the average blood sugar concentration over prolonged periods of time. In this work, an enzymatic colorimetric method for the measurement of HbA1c concentration of blood was developed, FPO (fructosyl peptide oxidase) enzyme was used to oxidize fructosyl valyl histidine released from neutral protease hydrolyzed HbA1c. The oxidation produces hydrogen peroxide which was further reacted with DA-64 dye by peroxidase (horseradish peroxidase, HRP) to develop color.
The optimum temperature for neutral protease to fully hydrolyze HbA1c was determined to be 60℃. One and half hours is required to have HbA1c been completely hydrolyzed when 40 KU/mL of neutral protease was employed. HbA1c concentration range from 5.2% to 15% can be detected. In order to reuse FPO enzyme, FPO was immobilized onto polydopamine modified PE membranes. It was found that further modified with glutaraldehyde (GA) on polydopamine-PE membranes could significantly enhance the immobilization efficiency of FPO. The stability of the immobilized FPO could also be improved that 95% activity still can be maintained after 7 consecutive reuse. The detection range of fructosyl valine by using immobilized FPO-PE membranes was 0.02-0.7mM.

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