Li Zhi-Zhang, Gong Fu-Chun, Shen Guo-Li, Yu Ru-Qin
State Key Laboratory for Chemo/Biosensing and Chemometrics, College of Chemistry and Chemical Engineering, Hunan University, Changsha, P R China.
Anal Sci. 2002 Jun;18(6):625-30. doi: 10.2116/analsci.18.625.
A novel amperometric immunosensor setup is described which uses horseradish peroxidase (HRP) as a label in conjunction with a current-based Brucella sensor. The Bacteria modified immunosensor was constructed by using a biocomposite formed by dispersing graphite powder into a mixture of Brucella melitensis and silicate polymer gel. The enzyme-labeled antibody can readily diffuse toward the encapsulated antigen (Brucella melitensis), which retains its binding properties, and the association reaction is easily detected at the surface exposed to the solution. The use of an oaminophenol (o-AP) substrate and amperometric detection at -150 mV (vs. SCE) results in a relatively low detection limit of 3.5 ng/ml and a linear detection range of 3.5 ng/ml to 200 ng/ml. Based on an optimized parameter, the prepared sensor was used to detect the Brucella melitensis antibody in serum samples by using a competitive binding assay. The results demonstrate the feasibility of employing the proposed immunosensor for the detection for Brucella melitensis antibody in a clinical analysis.
本文描述了一种新型的安培免疫传感器装置，该装置使用辣根过氧化物酶（HRP）作为标记物，并结合基于电流的布鲁氏菌传感器。通过将石墨粉分散到布鲁氏菌和硅酸盐聚合物凝胶的混合物中形成生物复合材料，构建了细菌修饰的免疫传感器。酶标记抗体能够轻松地向被包封的抗原（布鲁氏菌）扩散，该抗原保留其结合特性，并且在暴露于溶液的表面能够轻松检测到结合反应。使用邻氨基酚（o-AP）底物并在-150 mV（相对于饱和甘汞电极）下进行安培检测，检测限相对较低，为3.5 ng/ml，线性检测范围为3.5 ng/ml至200 ng/ml。基于优化参数，通过竞争结合试验，使用制备的传感器检测血清样本中的布鲁氏菌抗体。结果证明了在临床分析中使用所提出的免疫传感器检测布鲁氏菌抗体的可行性。
