Measurement of glucose concentrations in human plasma using a glucose biosensor.
Measurement of glucose concentrations in human plasma using a glucose biosensor.
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DOI：
10.1016/j.ab.2005.01.025
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发表时间：
2005-05
影响因子：
2.9
通讯作者：
Baoli Wu;Guomei Zhang;Yan Zhang;S. Shuang;Martin M. F. Choi
中科院分区：
文献类型：
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作者：
Baoli Wu;Guomei Zhang;Yan Zhang;S. Shuang;Martin M. F. Choi
葡萄糖是动植物碳水化合物的主要成分。人体血液中葡萄糖的定量测定对糖尿病的诊断和有效治疗具有重要意义。因此，相当大的努力集中在开发有效的诊断工具，以造福于糖尿病患者。近年来，化学传感器和生物传感器的研究发展非常迅速，出现了许多葡萄糖生物传感器[1-5]。然而，许多实验装置都很复杂，需要体积庞大且价格昂贵的电化学系统来进行循环伏安研究、电聚合和电沉积或用于光学传感的荧光分光计。因此，很明显，以更简单的设计和相对较低的成本改进葡萄糖生物传感器仍然是一个利基市场。在本工作中，我们尝试以蛋壳膜为酶固定化平台，以溶氧电极为传感器，构建葡萄糖生物传感器。蛋壳膜是蛋壳内的一层淡粉色双层膜，由类似角蛋白、胶原蛋白和弹性蛋白的高度交联的蛋白质组成。蛋壳膜具有良好的透气性和透水性，与酶具有良好的生物相容性，是酶固定化的理想生物平台[6-13]。此外，蛋壳膜比其他膜材料更便宜、更容易获得。尽管蛋壳膜与聚合物膜相比机械强度较低，但在保持湿润的情况下仍能提供良好的弹性和柔韧性。此外，我们发现使用钳子和剪刀并不是太难处理。蛋壳膜的成本最低。酶固定化过程的唯一主要成本是葡萄糖氧化酶(GOX)1和戊二醛。在商业和工业中，蛋壳膜肯定比现有的氧电极膜便宜。葡萄糖氧化酶最初固定在新鲜蛋壳膜的表面。酶固定化膜随后被放置在溶解氧传感器的表面。鉴于许多研究小组已经使用克拉克型电极来检测葡萄糖在酶转化过程中氧浓度的变化[14，15]，本文报道了一种在传统克拉克型氧电极的基础上发展起来的廉价的、改进的生物传感器来检测人血浆中的葡萄糖含量。该检测方案是基于葡萄糖的酶促反应导致基液中溶解氧水平的耗尽，然后监测氧水平的下降并与葡萄糖浓度相关联。该葡萄糖生物传感器具有稳定性好、成本低、设计简单、操作方便等特点。用于人血浆中葡萄糖含量的测定，结果令人满意。在蛋白和蛋黄被去除后，从一个破碎的新鲜蛋壳中小心地剥离一层蛋壳膜。它被大量的去离子水清洗过。将膜放置在干净的手表玻璃中，切成直径约15 mm的圆形；在pH 7.0的条件下，加入100L的0.8%(w/v)GOX溶液，
Glucose is a major component of animal and plant carbohydrates. Quantitative determination of glucose in human blood is of paramount importance for the diagnosis and effective treatment of diabetes. Therefore, considerable effort has been focused on developing effective diagnostic tools for the benefit of diabetic patients. In recent years, research in the development of chemosensors and biosensors has been very fast growing and many glucose biosensors have been presented [1–5]. However, many of the experimental setups are complicated and require a bulky and expensive electrochemical system for cyclic voltammetric studies, electropolymerization, and electrodeposition or spectrofluorimeter for optical sensing. Thus, it is obvious that there is still a niche for improving glucose biosensors with simpler design and relatively low cost. In this work, we attempted to construct a glucose biosensor using an eggshell membrane as the enzyme immobilization platform and a dissolved oxygen electrode as the transducer. Eggshell membrane is a light pink double-layered membrane inside the eggshell composed of highly cross-linked proteins similar to keratin, collagen, and elastion. Because of its excellent gas and water permeability and good biological compatibility with the enzyme, eggshell membrane may be an ideal bioplatform for enzyme immobilization [6–13]. Moreover, eggshell membrane is cheaper and easier to get than other membrane materials. Even though eggshell membrane lacks mechanical strength compared with polymeric membrane, it can still provide good elasticity and flexibility when it is kept wet. In addition, we find that it is not too diYcult to handle using forceps and scissors. The cost of eggshell membrane is minimal. The only major cost of the enzyme immobilization process is glucose oxidase (GOx) 1 and glutaraldehyde. Eggshell membrane is certainly cheaper than the existing membrane for oxygen electrodes in commerce and industry. Glucose oxidase was initially immobilized onto the surface of a fresh eggshell membrane. The enzymeimmobilized membrane was subsequently positioned on the surface of a dissolved oxygen sensor. Since many research groups have already used Clark-type electrodes to detect the change of oxygen concentration during enzymatic conversion of glucose [14, 15], in this article we report a cheaper and improved biosensor which is developed from a traditional Clark-type oxygen electrode to detect the glucose content in human blood plasma. The detection scheme was based on the enzymatic reactions of glucose leading to the depletion in the dissolved oxygen level in the base solution and then the decrease in the oxygen level was monitored and related to the glucose concentration. The proposed glucose biosensor has the promising features of good stability, low cost, simple design, and ease of operation. It was successfully applied to determine glucose content in human plasma and the results were satisfactory. An eggshell membrane was carefully peeled from a broken fresh eggshell after the albumen and yolk had been removed. It was cleaned with a copious amount of deionized water. The membrane was placed in a clean watch glass and cut into a circle of a diameter of about 15 mm; 100L of 0.8%(w/v) GOx solution at pH 7.0 was