Electrochemical Micro-immunosensors Using Monoclonal Antibodies

本研究项目的目标是开发高功能和微型免疫传感器，并研究这种传感器的实际应用的基本方面。在本研究中，电聚合膜首次成功地作为功能材料应用于制备如下所述的电化学免疫传感器。电位型人绒毛膜促性腺激素（hCG）传感器：当酪胺（=对-（2-氨乙基）-苯酚）在低电流密度下从碱性乙醇溶液中聚合时（例如，60 μ A/cm^2）时，得到了无孔、低电导率的精细织构薄膜。用这种聚酪胺修饰电极（= 0.5mm，=1-5 mm），在其上固定抗hCG单克隆抗体（抗小鼠腹水中hCG- -亚基），制成了一种选择性、快速（< 5 min响应90%）和灵敏（检测限为1 ng/ml）的电位型hCG激素传感器。此外，使用另一种单克隆抗体固定化， 关于我们 以该电极为参比电极，取消非免疫化学反应，再次测定人血清中的hCG，其浓度降至1 ng/ml。首先在酸性溶液中在铂电极上形成多孔聚酪胺膜，然后在碱性醇溶液中在铂电极表面覆盖聚酪胺膜，得到聚酪胺双涂层电极，然后通过肽连接将抗IgG固定在其上。采用竞争性酶免疫分析技术，在约100 μ g/ml的浓度范围内测定IgG。10 μ g/ml至1 mg/ml。新型二极管型生物传感器：将聚吡咯或聚苯胺薄膜覆盖在金膜电极上，并与金膜电极隔开一个小间隙，由于聚吡咯或聚苯胺薄膜的导电性随薄膜氧化态的变化而变化，因此该传感器表现为二极管型。该装置被应用于开发新的生物传感器的流动注射系统。少

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