Electrochemical Micro-immunosensors Using Monoclonal Antibodies

使用单克隆抗体的电化学微免疫传感器

• 批准号: 61870104
• 负责人: TANIGUCHI Isao
• 金额: $ 3.78万
• 依托单位: Kumamoto University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Developmental Scientific Research
• 财政年份: 1986
• 资助国家: 日本
• 起止时间: 1986 至 1988
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-61870104/
• 关键词: Electrochemical Immunosensor; Monoclonal Antibody; Immnoglobulin G; Human Chorionic Gonadotropin (hCG); Electropolymerization; Polytyramine Thin Film; Modified Electrode; 酵素免疫測定; 電解電合; 差動出力; 微小電極; 免疫センサ; 電位応答; イムノグロブリンG; 性腺刺激ホルモン; 電導性薄膜

The objectives of the present research project have been to develop highly functional and micro immunosensors, and also to study fundamental aspects on such sensors for practical use. In the present study, electropolymerized films have,at the first time, been applied successfully as functional materials to prepare electrochemical immunosensors as described below.Potentiometric human chorionic gonadotropin (hCG) sensor: when tyramine (=p-(2-aminoethy1)-phenol) is polymerized from an alkline alcohol solution at a low current density (e.g., 60muA/cm^2), a pore-free and fine-textured film with low conductivity was obtained. Using this polytyramine modified electrode ( =0.5, =1-5 mm), on which monoclonal anti-hCG (against hCG- -subunit from mouse ascites fluid) was immobilized, a potentiometric hCG hormone sensor was prepared, which responded selctively, rapidly (< 5 min. for 90% response) and sensitively (detection limit of 1 ng/ml). also, using an another monoclonal antibody immobilized e … More lectrode as a reference one, non-immunochemical response was canceled and down to 1 ng/ml of hCG in a human serum was again determined.Highly sensitive enzyme immunosensor for human immunogloblin G (IgG): A porous polytyramine film was formed on a Pt electrode at first from an acid solution and the surface was further covered with a polytyramine film from alkaline alcohol solution to give a polytyramine doubly coated electrode, on which anti-IgG was then immobilizes through the peptide linkage. By means of the competitive enzyme immunoassay technique, IgG was determined in the concentration range of ca. 10 pg/ml to 1 mg/ml. For the present electrode, a mixture of 0.5%Tween-20, a biological detergent, and 1% bovine serum albumin (BSA) was effective to diminish nonspecific responses.New diode-type biosensors: Polypyrrole or polyaniline thin film was covered on gold film electrodes with a small gap; this device behaved as a diode becarse the conductivity of the polymer film changed with the oxidation state of the film. This device was applied to develop new biosensors for a flow injection system. Less

本研究项目的目标是开发高功能和微型免疫传感器，并研究此类传感器的实际应用的基本方面。在本研究中，电聚合薄膜首次成功地用作功能材料来制备电化学免疫传感器，如下所述。 电位人绒毛膜促性腺激素（hCG）传感器：当酪胺（=p-（2-氨基乙基1）-苯酚）在低电流密度（例如， 60muA/cm^2)，得到无孔、纹理细密、低电导率的薄膜。使用固定有单克隆抗 hCG（针对来自小鼠腹水的 hCG-亚基）的聚酪胺修饰电极（=0.5，=1-5 mm），制备了电位 hCG 激素传感器，该传感器选择性、快速（< 5 分钟，90% 响应）且灵敏（检测限为 1 ng/ml）。此外，使用另一种固定化电极的单克隆抗体作为参比电极，取消非免疫化学反应，并再次测定人血清中低至 1 ng/ml 的 hCG。 用于人免疫球蛋白 G (IgG) 的高灵敏度酶免疫传感器：首先用酸溶液在 Pt 电极上形成多孔聚酪胺薄膜，然后在 Pt 电极上形成多孔聚酪胺薄膜。 表面进一步覆盖碱性醇溶液的聚酪胺膜，得到聚酪胺双层涂层电极，然后通过肽键将抗IgG固定在其上。通过竞争酶免疫分析技术，在约浓度范围内测定了IgG。 10 皮克/毫升至 1 毫克/毫升。对于本发明的电极，0.5%Tween-20（一种生物洗涤剂）和1%牛血清白蛋白（BSA）的混合物可有效减少非特异性反应。新型二极管型生物传感器：聚吡咯或聚苯胺薄膜以小间隙覆盖在金膜电极上；该器件表现为二极管，因为聚合物薄膜的电导率随薄膜的氧化态而变化。该设备用于开发用于流动注射系统的新型生物传感器。较少的

项目成果

谷口功: 病態生理. 8. 243-251 (1989)

谷口功：病理生理学。8. 243-251 (1989)

Ichiro Tsuji: Biosensors,.

辻一郎：生物传感器。

Isao,Taniguchi: "A Ferrocene-Modified Carbon Fiber Microelectrode as a Potentiometric Sensor for Hydrogen Peroxide" Extended Abstract of the 172nd Meeting of the Electrochemical Society, held at Honolulu. 87-2. 2213 (1987)

Isao,Taniguchi：“作为过氧化氢电位传感器的二茂铁改性碳纤维微电极”在檀香山举行的电化学学会第 172 届会议的扩展摘要。

谷口功: 学術月報. 42. 64 (1989)

谷口功：学术月报。42. 64 (1989)。

Isao,Taniguchi: "The Potentiometric Immunosensor Based on a Phlytyramine Modefied Electrode" Review of Polarography. 32. 83 (1986)

Isao，Taniguchi：“基于苯酪胺修饰电极的电位免疫传感器”极谱评论。