A piezoelectric immunoassay based on self-assembled monolayers of cystamine and polystyrene sulfonate for determination of Schistosoma japonicum antibodies
A piezoelectric immunoassay based on self-assembled monolayers of cystamine and polystyrene sulfonate for determination of Schistosoma japonicum antibodies
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基于胱胺和聚苯乙烯磺酸自组装单层的压电免疫分析法测定日本血吸虫抗体
DOI：
10.1007/s00216-002-1389-5
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发表时间：
2002
影响因子：
4.3
通讯作者：
R. Yu
中科院分区：
文献类型：
--
作者：
Cunchang Wang;Hua Wang;Zhaoyang Wu;Guoli Shen;R. Yu
基于改进的半胱胺(Cys)和聚苯乙烯磺酸盐(PSS)自组装单分子膜(SAM)固定化策略，研制了一种检测兔血清中日本血吸虫抗体(SjAb)的压电式免疫传感器。首先将半胱氨酸自组装膜应用于晶体的金电极表面，作为带正电荷的碱，然后通过带负电荷的PSS层将日本血吸虫抗原(SjAg)静电固定在晶体上。用适当选择的封闭剂封闭牛血清白蛋白和正常兔血清(NRS)，可显著减少非特异性吸附，免疫传感器用于在优化的缓冲介质中添加免疫增强剂聚乙二醇(PEG)测定SjAb。实验表明，Cys-PSS吸附固定化SjAg的免疫活性或结合效率高于戊二醛(GLU)结合或直接附着固定化SjAg。所研制的免疫传感器具有良好的灵敏度和检出限，且压电石英晶体易于再生。对感染兔血清样品的分析结果表明，该免疫传感器在日本血吸虫感染的临床诊断中是一种有希望的定性和定量检测SjAb的方法。
A piezoelectric immunosensor based on an improved immobilization strategy combining self-assembled monolayers (SAM) of cystamine (Cys) and polystyrene sulfonate (PSS) has been developed for the determination ofSchistosoma japonicumantibodies (SjAb) in rabbit serum. Cys SAM were first applied to the gold electrode surface of the crystal, serving as a positively-charged base.Schistosoma japonicumantigen (SjAg) was then electrostatically immobilized on the crystal by means of a negatively-charged PSS layer. When sealed by use of an appropriately selected blocking reagent for BSA and normal rabbit serum (NRS), non-specific adsorption could be substantially reduced.The immunosensor was used to determine SjAb in optimized buffer medium with addition of poly(ethylene glycol) (PEG), which served as an immunoreaction enhancer. It was shown experimentally that SjAg immobilized by the Cys–PSS adsorption procedure had higher immunological activity or binding efficiency than those immobilized by the glutaraldehyde (GLU) binding or direct attachment procedures. The immunosensor developed had satisfactory sensitivity and detection limit, and regeneration of the piezoelectric quartz-crystal was easy. Analytical results obtained with infected rabbit serum samples indicated that the proposed immunosensor is a promising alternative for qualitative and quantitative determination of SjAb in clinical diagnosis of infection withSchistosoma japonicum.