An amperometric biosensor for acetylcholine determination prepared from acetylcholinesterase-choline oxidase immobilized in polypyrrole-polyvinylsulpfonate film
An amperometric biosensor for acetylcholine determination prepared from acetylcholinesterase-choline oxidase immobilized in polypyrrole-polyvinylsulpfonate film
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DOI：
10.1016/j.snb.2014.06.049
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发表时间：
2014-10-01
影响因子：
8.4
通讯作者：
Arslan, Fatma
中科院分区：
文献类型：
--
作者：
Aynaci, Elif;Yasar, Ahmet;Arslan, Fatma
本文以聚吡咯-聚乙烯基磺酸盐(PPy-PVS)膜为载体，研制了一种新型的电流型乙酰胆碱(ACh)生物传感器。以戊二醛(GA)和牛血清白蛋白(BSA)为交联剂，将乙酰胆碱酯酶(AChE)和胆碱酯酶(CHO)固定在PPy-PVS膜上。乙酰胆碱的测定是在0.3V与Ag/AgCl条件下用酶法产生的过氧化氢氧化完成的。考察了pH值和温度的影响，确定最佳工艺参数分别为9.0℃和65℃。在1.0×10(-8)-1.0×10(-7)M(R-2=0.994)和1.0×10(-7)-1.0×10(-6)M(R-2=0.994)范围内有两个线性部分。该生物传感器的检测下限为5.0×10~(-9)M，响应时间为200×10~(-9)M，并对酶电极的储存稳定性和操作稳定性进行了研究。结果表明，经18次活性测定后，响应电流的保留率为83.1%。ACh生物传感器检测结果重现性好，相对标准偏差为4.4%。制备的ACh生物传感器在4摄氏度的0.1M甘氨酸缓冲溶液中保存35天后，保持了31%的初始活性。(C)2014 Elsevier B.V.版权所有。
In this paper, a novel amperometric acetylcholine (ACh) biosensor with immobilization of acetylcholinesterase-choline oxidase (AChE-ChO) on polypyrrole-polyvinylsulphonate (PPy-PVS) film has been successfully developed. AChE and ChO enzymes were immobilized on PPy-PVS film by cross-linking with glutaraldehyde (GA) and bovine serum albumine (BSA). Determination of ACh was carried out by oxidation of enzymatically produced H2O2 at 0.3 V vs Ag/AgCl. The effects of pH and temperature were investigated and optimum parameters were found to be 9.0 and 65 degrees C, respectively. There were two linear parts in the region between 1.0 x 10(-8)-1.0 x 10(-7) M (R-2 = 0.994) and 1.0 x 10(-7)-1.0 x 10(-6) M (R-2=0.994). The detection limit of the biosensor was 5.0 x 10(-9) M and response time of the biosensor was 200 s. Storage stability and operation stability of the enzyme electrode were also studied. The results showed that 83.1% of the response current was retained after 18 activitiy assays. ACh biosensor gave perfect reproducible findings with 4.4% relative standard deviation. The prepared ACh biosensor retained 31% of its initial activity after 35 days storage in 0.1 M glycine buffer solution at 4 degrees C. (C) 2014 Elsevier B.V. All rights reserved.