石英晶体微天平传感的酶联免疫分析法测定免疫球蛋白IgM  

Quartz Crystal Microbalance ELISA for Determination of IgM

作　　者：周亚民[1] 王永东[1] 王桦[2] 吴朝阳[2] 沈国励[2] 

机构地区：[1]东莞理工学院化学生物工程系,广东东莞523106 [2]湖南大学化学生物传感与计量学国家重点实验室,长沙410082

出　　处：《传感技术学报》2006年第2期305-308,331,共5页Chinese Journal of Sensors and Actuators

基　　金：广东省教育厅自然科学研究项目资助(Z03093);国家自然科学基金资助(20375008);东莞市科技项目资助(2005054)

摘　　要：结合酶联免疫分析技术与石英晶体微天平检测技术,发展了一种基于石英晶体微天平传感的免疫传感器用于测定血液中免疫球蛋白IgM的量。羊抗人IgM抗血清通过氨基硅烷化共键固定在石英晶体表面上,采用竞争吸附免疫分析法,以辣根过氧化物酶(HRP)标记的IgM作标记物,N-四甲联苯胺(TMB)和H2O2作酶的底物。结合在石英晶体微天平上的酶标记物催化TMB氧化形成沉淀,使石英晶体微天平的振荡频率发生改变,据此测定标记物HRP-IgM和IgM的量。实验优化了分析条件下,对IgM的检测范围为0.03μg/mLto7μg/mL,它具有仪器简单、操作方便、灵敏度高等优点。Enzyme-linked immunosorbent assay （ELISA） is an extremely useful immunoassay method. The quartz crystal microbalance is highly sensitive to the surface mass change. The aim of this study was to develop an amplified mass enzyme linked immunosensor with quartz crystal microbalance as transducer to determine IgM. Sheep anti-human IgM antibody was covalently immobilized on transducer surface via the modified （γ-aminoppropyl） trimethoxysilane, a competitive immunoassay format was adopted with HRPIgM as a tracer, Tetramethyl Benzidine （TMB） and H2O2 as the enzyme substrates. TMB was catalyzed oxide to make deposition produce on quartz crystal microbalance surface. The amount of HRP-IgM binding onto the transducer surface was detected through measuring the resonance frequency change of quartz crystal microbalance. The system was optimized to realize a reliable determination of IgM in the range of 0.03μg/mL to 7μg/mL. It exhibits some advantages, such as simplicity of fabrication, rapidity of measurement, and satisfactory sensitivity.

关 键 词：酶联免疫分析法 石英晶体微天平 IgM的检测 

分 类 号：TP212.3[自动化与计算机技术—检测技术与自动化装置]