Bojorge-Ramírez Ninoska I, Salgado Andréa Medeiros, Valdman Belkis
Escola de Química, Centro de Tecnologia, Federal University of Rio de Janeiro, Rio de Janeiro, Brazil.
Assay Drug Dev Technol. 2007 Oct;5(5):673-82. doi: 10.1089/adt.2007.082.
An immunosensor for detecting the antibody anti-apyrase of Schistosoma mansoni based on rigid composite materials, containing graphite powder and epoxy resins, developed in this work, is described. A surface modification strategy for the use of oxidized graphite in the detection of antibody-antigen interaction was developed. This modification strategy is based on silanization of conductive composite. First, the graphite powder-epoxy resin was treated with concentrated hydrogen peroxide to improve surface hydroxyl groups and to form a hydrophilic layer. Second, 3- aminopropyltriethoxysilane was subsequently used to functionalize the treated surface to form amino groups, which were further activated with glutaraldehyde to introduce a layer of aldehyde groups. Contact angle microscopy and scanning electron microscopy were used as a qualitative analysis of the deposition of silane on the surface of the sensor. The effectiveness of the modification strategy was validated by amperometric immunoassays of S. mansoni. Amperometric signals related to concentrations of this immobilized protein were observed, and the effects of pH and incubation times were analyzed. This surface modification strategy provides a platform on which proteins can be directly immobilized for immunosensor and protein array applications.
本文描述了一种基于刚性复合材料（包含石墨粉和环氧树脂）开发的用于检测曼氏血吸虫抗腺苷三磷酸双磷酸酶抗体的免疫传感器。开发了一种在抗体 - 抗原相互作用检测中使用氧化石墨的表面修饰策略。这种修饰策略基于导电复合材料的硅烷化。首先，用浓过氧化氢处理石墨粉 - 环氧树脂，以增加表面羟基并形成亲水层。其次，随后使用3 - 氨丙基三乙氧基硅烷对处理过的表面进行功能化以形成氨基，再用戊二醛进一步活化以引入一层醛基。使用接触角显微镜和扫描电子显微镜对硅烷在传感器表面的沉积进行定性分析。通过曼氏血吸虫的安培免疫测定验证了修饰策略的有效性。观察到与这种固定化蛋白质浓度相关的安培信号，并分析了pH值和孵育时间的影响。这种表面修饰策略提供了一个平台，可直接在其上固定蛋白质用于免疫传感器和蛋白质阵列应用。
