Development of choline and acetylcholine Pt microelectrodes.
Development of choline and acetylcholine Pt microelectrodes.
复制标题
胆碱和乙酰胆碱铂微电极的开发。
DOI：
10.1006/abio.1993.1550
复制
发表时间：
1993
影响因子：
2.9
通讯作者：
Guilbault,GG
中科院分区：
文献类型：
--
作者：
Huang,Z;Villarta-Snow,R;Lubrano,GJ;Guilbault,GG
将胆碱氧化酶（ChO）和乙酰胆碱酯酶（AchE）固定在玻璃密封的25 μm铂丝的尖端，研制了胆碱（Ch）和乙酰胆碱（Ach）微酶传感器。测试了几种固定方法，包括电聚合物对酶/s的编码定位和与戊二醛的交联。所使用的各种电聚合物有1,2-二氨基苯、间苯二酚、4-羟基苯磺酸以及1,2-二氨基苯和间苯二酚两种聚合物的组合。由醋酸纤维素（CA）构建的内膜在戊二醛固定之前沉积。确定了微电极的分析特性，包括固定程序的优化、校准曲线、pH响应曲线、稳定性和对脑细胞外液中可能存在的电活性化合物的选择性。将酶与戊二醛交联在CA内膜上制备最佳微电极。在5.0 × 10−7 ~ 1.0 × 10−4M Ch和5.0 × 10−7 ~ 9.3 × 10−5M Ach浓度范围内，响应呈线性关系。达到95%稳态电流的时间为15 ~ 20 s。ca包覆的Ch微电极可用于测量人工脑胞外液中Ch浓度的变化。
Choline (Ch) and acetylcholine (Ach) microenzyme sensors were developed based on the immobilization of choline oxidase (ChO) and acetylcholinesterase (AchE) at the tip of a 25-μm Pt wire sealed in glass. Several immobilization procedures were tested, including code-position of the enzyme/s with an electropolymer and cross-linking with glutaraldehyde. The various electropolymers used were 1,2-diaminobenzene, resorcinol, 4-hydroxybenzenesulfonic acid, and a combination of two polymers, 1,2-diaminobenzene and resorcinol. An inner membrane constructed from cellulose acetate (CA) was deposited prior to immobilization with glutaraldehyde. The analytical characteristics of the microelectrodes, including optimization of immobilization procedures, calibration curves, pH response curves, stability, and selectivity toward possible electroactive compounds found in the brain extracellular fluid, were determined. The best microelectrodes were prepared by cross-linking the enzymes with glutaraldehyde on top of the inner CA membrane. The responses are linear in the concentration range 5.0 × 10−7-1.0 × 10−4M Ch and 5.0 × 10−7-9.3 × 10−5M Ach. The time to reach 95% steady-state current was 15-20 s. The CA-coated Ch microelectrodes were useful for measurement of changes in Ch concentration in artificial brain extra-cellular fluid.