| Summary: | 碩士 === 國立臺灣科技大學 === 化學工程系 === 102 === 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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