CONDUCTING POLYMER ENZYME BASED BIOSENSOR FOR NO

传导基于聚合物酶的生物传感器

• 批准号: 6564505
• 负责人: Hiroyasu Tachikawa
• 金额: $ 25.93万
• 依托单位: JACKSON STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-02-01 至 2005-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6564505
• 关键词: Raman spectrometry; aging; amines; bioengineering /biomedical engineering; biomedical equipment; biosensor device; biotechnology; cytochrome c; dopamine; electrochemistry; electron transport; glucose; hemoprotein; immobilized enzymes; infrared spectrometry; interferometry; laboratory rat; microelectrodes; nitric oxide; polymers; respiratory gas analyzer; tissue /cell culture; ultraviolet spectrometry

Conducting polymers have been used to modify the electrode surface and to immobilize the enzymes to construct biosensors for the detection of biomedically important species such as glucose, nitric oxide, histamine, etc. We will continue to use this approach to develop bioelectrodes which can catalytically reduce biomedically relevant species, and eventually develop a sensitive transducer for the detection of those species. For the detection of NO we will construct microsensors using both conducting polymers, and hemoprotein enzymes. Hemoproteins including hemoglobin (Hb), myoglobin (Mb), and cytochrome c peroxidase as well as the mutants of these proteins will be used to modify the surface of microelectrodes for an effective catalytic reduction of NO at less negative potentials that should minimize the effects of such interferents as nitrite, nitrite, and ascorbic acid. Since this type of electrodes use reductive potentials, the effects of catecholamines on the detect of NO can be eliminated. Conducting polymers using both polyhemin and polyhematin will be electrochemically coated on carbon fiber electrodes for the preparation of microsensors of the determination of NO. The utility of these electrodes will be tested in cell cultures. Spectroscopic methods including laser-Raman, FT-IR, and UV-visible absorption will be used to study the conformation of protein films on the electrode and also to elucidate the structural changes occurring under the different conditions including applied potential, pH, and in the presence of such species as NO and O2. We also plan to construct electrochemical amine sensors which are based on tryptophan tryptophylquinone (TTQ)- containing quinoproteins such as methylamine dehydrogenase (MADH) and aromatic amine dehydrogenase (AADH). Conducting polymers such as polypyrrole (PPy) will be used as a matrix to immobilize these enzymes on gold electrodes. These sensors will be characterized for the detection of dopamine, histamine, and other primary aliphatic and aromatic amines.

导电聚合物已被用来修改的电极表面，并将酶构建生物传感器的检测生物医学上重要的物种，如葡萄糖，一氧化氮，组胺等，我们将继续使用这种方法来开发生物电极，可以催化减少生物医学相关的物种，并最终开发一个敏感的传感器，用于检测这些物种。为了检测NO，我们将使用导电聚合物和血红素蛋白酶构建微传感器。血红蛋白，包括血红蛋白（Hb），肌红蛋白（Mb），细胞色素c过氧化物酶，以及这些蛋白质的突变体将被用来修改微电极的表面，有效的催化还原NO在较低的负电位，应尽量减少这种干扰的影响，如亚硝酸盐，亚硝酸盐，和抗坏血酸。由于这种电极使用还原电位，可以消除儿茶酚胺对NO检测的影响。导电聚合物，使用多氯化血红素和多氯化血红素将电化学涂覆在碳纤维电极上的微传感器的测定NO的制备。这些电极的效用将在细胞培养物中进行测试。光谱方法，包括激光拉曼，FT-IR和紫外-可见吸收将被用来研究构象的蛋白质膜上的电极，也阐明了结构的变化发生在不同的条件下，包括施加的电位，pH值，并在存在的物种，如NO和O2。我们还计划构建电化学胺传感器，这是基于含有色氨酸的醌（TTQ）的醌蛋白，如甲胺脱氢酶（MADH）和芳香胺脱氢酶（AADH）。导电聚合物如聚吡咯（PPy）将被用作基质，将这些酶固定在金电极上。这些传感器的特点是检测多巴胺，组胺，和其他主要的脂肪族和芳香族胺。