Immunosensor for Mycobacterium tuberculosis on screen-printed carbon electrodes
Immunosensor for Mycobacterium tuberculosis on screen-printed carbon electrodes
复制标题
DOI：
10.1016/j.bios.2004.09.035
复制
发表时间：
2005-04-15
影响因子：
12.6
通讯作者：
Costa-García, A
中科院分区：
文献类型：
--
作者：
Díaz-González, M;González-García, MB;Costa-García, A
在这项工作中，两种方法进行了比较，以生产酶伏安免疫传感器的结核分枝杆菌抗原（Ag 360和Ag 231）的测定，使用预氧化丝网印刷碳电极（SPCE）作为信号转导元件。碱性磷酸酶（AP）与底物3-吲哚酚磷酸（3-IP）联合使用。结核抗原和抗M.在电极表面形成结核。然后用生物素标记的兔抗M抗体捕获免疫复合物。结核抗体，通过链霉亲和素：生物素反应固定在链霉亲和素修饰的SPCE上。最后，用碱性磷酸酶（AP）标记的兔抗鼠IgG作为检测抗体。结核抗原被抗M的单克隆抗体捕获。结核杆菌，其通过与被动吸附在SPCE上的兔IgG反应而固定在电极表面上。生物素标记的兔抗M.结核抗体与碱性磷酸酶标记的链霉亲和素一起用作检测抗体。用链霉亲和素修饰的SPCE和抗原Ag 231与单克隆抗体MabF 184-3的免疫复合物制成的免疫传感器检测结核抗原，检测限为1.0ng/ml。免疫传感器也适用于Ag 231加标蛋白质基质。(c)2004 Elsevier B. V.保留所有权利。
In this work, two methods have been compared to produce enzymatic voltammetric immunosensors for the determination of Mycobacterium tuberculosis antigens (Ag360 and Ag231), using a pre-oxidised screen-printed carbon electrode (SPCE) as a signal transduction element. The enzyme alkaline phosphatase (AP) was used in combination with the substrate 3-indoxyl phosphate (3-IP).In one design, the immune complexes between M. tuberculosis antigens and monoclonal antibodies against M. tuberculosis were formed out of the electrode surface. Then, the immune complexes were captured by biotinylated rabbit anti-M. tuberculosis antibodies, immobilised on the streptavidin modified SPCEs through the streptavidin:biotin reaction. Finally, an alkaline phosphatase (AP) labelled rabbit IgG anti-mouse immunoglobulin G was used as a detector antibody.In the other design, the M. tuberculosis antigens were captured by monoclonal antibodies against M. tuberculosis, which were immobilised on the electrode surface through the reaction with rabbit IgG passively adsorbed on the SPCEs. The biotinylated rabbit anti-M. tuberculosis antibodies were used with an alkaline phosphatase labelled streptavidin as detector antibodies.The best results for M. tuberculosis antigen determination were obtained using the immunosensor on the streptavidin modified SPCEs and the immune complexes between antigen Ag231 and monoclonal antibodies MabF 184-3, with a detection limit of 1.0 ng/ml. The immunosensor was also applied to Ag231 spiked proteic matrices. (c) 2004 Elsevier B.V. All rights reserved.