Gong Fu-Chun, Tang Lian-Hui, Shen Guo-Li, Yu Ru-Qin
State Key Laboratory for Chemo/Biosensing and Chemometrics, College of Chemistry and Chemical Engineering, Hunan University, Changsha 410082, PR China.
Talanta. 2004 Mar 10;62(4):735-40. doi: 10.1016/j.talanta.2003.09.023.
A fluoroimmunosensing device which was based on ferulic acid (FA)/horseradish peroxidase system for the detection of Schistosoma japonicum antibody (SjAb) has been developed. To circumvent the difficulty of regeneration of immunocomposite surface, a natural chitosan-epoxy resin matrix was used for the immobilization of SjAg. The surface of the immunocomposite layer reacted was easily regenerated by simple polishing. The renewed surface served as a platform for the competitive immuno-reaction of HRP-SjAb and SjAb with SjAg immobilized at the support body surface and for enzymatic reaction. A novel fluorescent substrate ferulic acid for HRP, which is relatively stable toward H(2)O(2), has been adapted in the proposed fluorometric enzyme immunosensing system. FA can been catalyzed to produce a non-fluorescent species. The amount of HRP-SjAb bound to the aforementioned renewable surface layer, which is related to the content of SjAb in samples could be quantitized by measuring the decrease of fluorescence of FA induced by HRP-SjAb. The chitosan incorporated in matrix is favorable for the amplification of this sensing system due to the electrostatic reaction with FA. The proposed method showed a linear response ranging from 45 to 150ngml(-1), with an improved detection limit of 45ngml(-1). The method has been employed to determine SjAb in serum samples.
一种基于阿魏酸（FA）/辣根过氧化物酶体系用于检测日本血吸虫抗体（SjAb）的荧光免疫传感装置已被开发出来。为了克服免疫复合物表面再生的困难，使用天然壳聚糖 - 环氧树脂基质固定日本血吸虫抗原（SjAg）。免疫复合物反应层的表面通过简单抛光很容易再生。更新后的表面作为辣根过氧化物酶标记的日本血吸虫抗体（HRP - SjAb）和SjAb与固定在支撑体表面的SjAg进行竞争性免疫反应以及酶促反应的平台。一种对过氧化氢相对稳定的新型辣根过氧化物酶荧光底物阿魏酸已应用于所提出的荧光酶免疫传感系统中。FA可被催化产生一种非荧光物质。结合到上述可再生表面层的HRP - SjAb的量与样品中SjAb的含量相关，通过测量HRP - SjAb诱导的FA荧光降低可对其进行定量。由于壳聚糖与FA的静电反应，基质中掺入的壳聚糖有利于该传感系统的信号放大。所提出的方法在45至150 ng ml⁻¹范围内呈线性响应，检测限提高到45 ng ml⁻¹。该方法已用于测定血清样品中的SjAb。
