Characteristics of the Multifunctional Biosensors for the Measurements of Bioactive Substances in the Living Body

用于测量活体内生物活性物质的多功能生物传感器的特点

• 批准号: 05558115
• 负责人: SHIMIZU Nobuaki
• 金额: $ 10.56万
• 依托单位: Kanazawa University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Developmental Scientific Research (B)
• 财政年份: 1993
• 资助国家: 日本
• 起止时间: 1993 至 1995
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-05558115/
• 关键词: biosensor; immunosensor; immobilized enzyme; quartz crystal sensor; bioacitive substance; immunosorbent assay; glucose oxidase; microdialysis sensor; 固定化抗体; 抗原・抗体反応; ヒトIgG; 抗ヒトIgG; マイクロバランス; バイオセンサー; 免疫センサー; 酵素センサー; ボルタンメトリー; 単一ニューロン活動

The purpose of this research project was to accomplish the biosensors which can detect the endogenous substances influencing neuronal activity in the brain. For this purpose, the characteristics of the following sensors were studied ; (1) microdialysis sensor for separation and analysis of bioactive substances, (2) enzyme biosensor for detection of glucose immobilized with glucose oxidase and (3) a quartz crystal microbalance sensor for immunosorbent assay.1. Microdialysis biosensorNoradrenaline release in the medial prefrontal cortex and spleen of rats was studied. The detection limit of the microdialysis sensor system for noradrenaline was 500 fg/40 mul at a 3 : 1 signal to noise ratio. A series of experiments using pharmacological and physiological manipulations indicated that the noradrenaline recovered in the dialysate was mainly derived from nerve terminals.2. Enzyme biosensorGlucose oxidase was immobilized onto platinum electrode to attain stabilized activity. The concentration … More response characteristics and the stability of the sensor were studied for glucose concentrations of 0.1-10.0mM.Steady state response of the sensor was proportional to the glucose concentration up to 5 mM.The slope of the linear correlation of sensor response was inversely proportional to the dissolved oxygen concentration. The decrease in sensor response caused by the reduction of dissolved oxygen concentration was overcome by using ferrocene carboxialdehyde as an electron mediator.3. Quartz crystal microbalance sensorDetermination of human immunoglobulin G (IgG) was performed by using a piezoelectric crystal biosensor system, which was consist of an AT-cut quarts piezoelectriccrystal, oscillator and frequency counter. Anti-human IgG was immobilized onto the surface of the gold electrode with tyramine and glutaraldehyde. The resonant frequency shift resulting from the affinity reaction of anti-IgG and IgG correlated with a concentration of IgG in the range of 10mug/ml-50mug/ml. The crystal sensor showed no response to bovine serum albumin. Less

本课题的目的是研制能够检测脑内影响神经元活动的内源性物质的生物传感器。为此，我们研究了以下传感器的特性：（1）用于生物活性物质分离和分析的微透析传感器，（2）用于葡萄糖检测的固定化葡萄糖氧化酶酶生物传感器和（3）用于免疫吸附测定的石英晶体微天平传感器.微透析生物传感器研究了大鼠内侧前额叶皮层和脾脏去甲肾上腺素释放的变化。在3：1的信噪比下，微透析传感器系统对去甲肾上腺素的检测限为500 fg/40穆尔。一系列的药理和生理操作实验表明，透析液中回收的去甲肾上腺素主要来源于神经末梢.酶生物传感器将葡萄糖氧化酶固定在铂电极上，使酶活性稳定.浓度 ...更多信息 在0.1- 10.0mM的葡萄糖浓度范围内，研究了传感器的响应特性和稳定性，传感器的稳态响应与葡萄糖浓度成正比，直至5 mM，传感器响应的线性相关斜率与溶解氧浓度成反比。以二茂铁甲醛为电子媒介，克服了溶解氧浓度降低引起的传感器响应下降.石英晶体微天平传感器采用AT型石英晶体压电晶体、振荡器和频率计数器组成的压电晶体生物传感器系统测定人免疫球蛋白G（IgG）。用酪胺和戊二醛将抗人IgG固定在金电极表面。由抗-IgG和IgG的亲和反应产生的共振频移与IgG浓度在10 μ g/ml-50 μ g/ml范围内相关。该晶体传感器对牛血清白蛋白无响应。少

项目成果

清水宣明: "拘束ストレスと摂食抑制の中枢機序" Neurosciences. (印刷中). (1995)

N. Shimizu：“束缚应激和进食抑制的中心机制”，《神经科学》（出版中）。

Aoyagi,K.: "Concurrent measurement of serotonin metabolism and single neuron activity changes in the lateral hypothalamus of freely behaving rat." Behav.Brain Res.49. 205-212 (1992)

Aoyagi,K.：“同时测量自由行为大鼠下丘脑外侧的血清素代谢和单个神经元活动变化。”

Hori, T.et al.: "The hypothalamo-sympathetic nervous system modulates peripheral cellular immunity." Neurobiology. (in press). (1996)

Hori, T.et al.：“下丘脑交感神经系统调节外周细胞免疫。”

Nakane,H: "Stress-induced noradrenaline release in the rat prefrontal cortex measured by microdialysis" Am.J.Physiol.267. R1559-R1566 (1994)

Nakane,H：“通过微透析测量大鼠前额皮质中压力诱导的去甲肾上腺素释放”Am.J.Physiol.267。

Lee,R.-S.: "Effects of systemic and local ethanol onresponses of rat cerebellar Purkinje neurons to iontophoretically applied γ-aminobutyric acid" Brain Res.687. 1-11 (1995)

Lee, R.-S.：“全身和局部乙醇对大鼠小脑浦肯野神经元对离子电渗疗法应用的 γ-氨基丁酸的反应的影响”Brain Res.687 (1995)。