摘要
同时固定四羧基酞菁钻（Ⅲ）（CoPc）和HRP标记核基质蛋白22抗体（HRP-Ab-NMP22）于自组装在金电极表面的Fe3O4／Au胶上，构建了一种快速测定膀胱肿瘤患者尿液中NMP22抗原（NMP22）含量的安培免疫传感器。CoPc可被用作电檄表面HRP的电子传递媒介体。当该传感器在含NMP22尿样的溶液中温育后，NMP22与HRP-Ab-NMP22免疫结合导致HRP的活性中心与CoPc之间的电子传递部分阻碍，使HRP对H202电催化还原电流Io，降低。△Io与NMP22浓度在1．2～200μg／L呈线性关系；检出限为0．5μg／L。该传感器对NMP22响应灵敏，较ELISA法提高了检测速度，有望用于膀胱肿瘤的体外诊断。
A novel immunosensor was proposed for the rapid determination without separation of nuclear matrix protein 22 （NMP22） antigen in human urine . The immunosensor was prepared by co-immobilizing hoseradish peroxidase （HRP）-NMP22 antibody, Co（Ⅲ） phthalocyanine （CoPc） on a Fe3O4/Au collide -modified gold electrode （HRP-Ab-NMP22/ Fe3O4/Au colloid/CoPc CME） using the self-assembly technique in which CoPc was as electron immediate. The modified electrode shows an excellent electrocatalytic activity for NMP22 antigen in phosphate buffer solution （ pH 7.0）. After the immunosensor is incubated with NMP22 antigen solution at 37℃ for 20 min, the access of activity center of the HRP to electrode is partly inhibited, which leads to a linear decrease of the catalytic efficiency of the HRP to the reduction of immobilized CoPc by H2O2 at -50 mV in NMP22 antigen concentration ranges from 1.2 to 200 μg/L. The detection limit was 0.5 μg/L. The immunosensor was successfully utilized for the determination of NMP22 antigen in urine samples of bladder patients, which shows a good accuracy, precision and reproducibility. This method decreases the detection cost and shortens the analytical time, thus would be valuable for clinical immunoassay for bladder cancer.
出处
《分析化学》
SCIE
EI
CAS
CSCD
北大核心
2007年第11期1553-1558,共6页
Chinese Journal of Analytical Chemistry
基金
浙江省自然科学基金(Y106725)
浙江省科技攻关项目(No2006C31040)
浙江省教育厅基金(20061697)
宁波市自然科学基金(No2006A610048)资助项目
