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，并研究这些传感器的基本方面以供实际使用。在本研究中，电聚合膜首次成功地作为功能材料用于制备电化学免疫传感器，如下所述。电位法人绒毛膜促性腺激素（hCG）传感器：当从碱性酒精溶液中以低电流密度（例如60muA/cm^2）聚合酪胺（=p-(2-氨基乙基1)-苯酚）时，获得无孔、细纹理、低电导率的薄膜。采用聚酪胺修饰电极（=0.5,=1-5 mm），固定化单克隆抗hCG（针对小鼠腹水hCG- -亚基），制备了具有选择性、快速（< 5 min， 90%应答）、灵敏度高（检测限为1 ng/ml）的hCG激素传感器。同时，用另一种单克隆抗体固定e . More电极作为参考电极，取消非免疫化学反应，再次测定人血清中低至1ng /ml的hCG。人体免疫球蛋白G （IgG）高灵敏度酶免疫传感器：首先用酸溶液在铂电极上形成多孔的聚酪胺膜，再用碱醇溶液中的聚酪胺膜覆盖表面，得到双聚酪胺包被电极，然后通过肽链固定抗IgG。采用竞争酶免疫分析法测定IgG，浓度范围约为10pg /ml ~ 1mg /ml。对于目前的电极，0.5%吐温-20，生物洗涤剂和1%牛血清白蛋白（BSA）的混合物可以有效地减少非特异性反应。新型二极管型生物传感器：将聚吡咯或聚苯胺薄膜覆盖在间隙较小的金膜电极上；该装置表现为二极管，因为聚合物薄膜的电导率随着薄膜的氧化状态而变化。该装置已被应用于开发用于流动注射系统的新型生物传感器。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