Sassolas Audrey, Blum Loïc J, Leca-Bouvier Béatrice D
Institut de Chimie et Biochimie Moléculaires et Supramoléculaires, Laboratoire de Génie Enzymatique et Biomoléculaire, Université de Lyon, Université Lyon 1, CNRS, UMR5246, 69622, Villeurbanne Cedex, France.
Anal Bioanal Chem. 2009 Jun;394(4):971-80. doi: 10.1007/s00216-009-2780-2. Epub 2009 Apr 24.
Performant reagentless electrochemiluminescent (ECL) (bio)sensors have been developed using polymeric luminol as the luminophore. The polyluminol film is obtained by cyclic voltammetry (CV) on a screen-printed electrode either in a commonly used H(2)SO(4) medium or under more original near-neutral buffered conditions. ECL responses obtained after performing polymerization either at acidic pH or at pH 6 have been compared. It appears that polyluminol formed in near-neutral medium gives the best responses for hydrogen peroxide detection. Polymerization at pH 6 by cyclic voltammetry gives a linear range extending from 8 x 10(-8) to 1.3 x 10(-4) M H(2)O(2) concentrations. Based on this performant sensor for hydrogen peroxide detection, an enzymatic biosensor has been developed by associating the polyluminol film with an H(2)O(2)-producing oxidase. Here, choline oxidase (ChOD) has been chosen as a model enzyme. To develop the biosensor, luminol has been polymerized at pH 6 by CV, and then an enzyme-entrapping matrix has been formed on the above modified working electrode. Different biological (chitosan, agarose, and alginate) and chemical (silica gels, photopolymers, or reticulated matrices) gels have been tested. Best performances have been obtained by associating a ChOD-immobilizing photopolymer with the polyluminol film. In this case, choline can be detected with a linear range extending from 8 x 10(-8) to 1.3 x 10(-4) M.
已经开发出了使用聚鲁米诺作为发光体的高性能无试剂电化学发光（ECL）（生物）传感器。聚鲁米诺膜是通过循环伏安法（CV）在丝网印刷电极上，在常用的硫酸（H₂SO₄）介质中或在更原始的近中性缓冲条件下获得的。比较了在酸性pH或pH 6下进行聚合后获得的ECL响应。结果表明，在近中性介质中形成的聚鲁米诺对过氧化氢检测给出了最佳响应。通过循环伏安法在pH 6下聚合得到了一个线性范围，从8×10⁻⁸到1.3×10⁻⁴ M的过氧化氢浓度。基于这种用于过氧化氢检测性能优异的传感器，通过将聚鲁米诺膜与产生过氧化氢的氧化酶结合，开发出了一种酶生物传感器。在这里，胆碱氧化酶（ChOD）被选作模型酶。为了开发该生物传感器，鲁米诺已通过CV在pH 6下聚合，然后在上述修饰的工作电极上形成了一个酶包埋基质。已经测试了不同的生物（壳聚糖、琼脂糖和藻酸盐）和化学（硅胶、光聚合物或网状基质）凝胶。通过将固定ChOD的光聚合物与聚鲁米诺膜结合获得了最佳性能。在这种情况下，可以检测胆碱，线性范围从8×10⁻⁸到1.3×10⁻⁴ M。
