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布鲁氏菌颗粒在光纤上的固定化策略及其在化学发光免疫传感器中的应用。

Immobilization strategies of Brucella particles on optical fibers for use in chemiluminescence immunosensors.

机构信息

Department of Biotechnology Engineering, Ben-Gurion University of the Negev, Beer-Sheva 84105, Israel.

出版信息

Talanta. 2009 Nov 15;80(1):338-45. doi: 10.1016/j.talanta.2009.06.070. Epub 2009 Jul 7.

Abstract

Immunosensors are powerful analytical tools in clinical and veterinary diagnostics. This has led us to design a chemiluminescent immunosensor aimed at identifying anti-Brucella antibodies using optical fibers as the transducer. In order to develop the optimal transducer, to achieve an optimal chemical modification thereby allowing an optimal covalent binding of the protein receptor, several cleaning strategies and silane coupling agents were investigated. Brucella killed organisms were used as a model receptor for quantifying anti-Brucella IgG antibodies in a suspension compared to conventional colorimetric and chemiluminescent ELISA. A silane-benzophenone derivative was selected as the best performing silane coupling agent: the optical fiber immunosensor (OFIS) has showed the lowest limit of detection at 0.207 microg/ml, compared to 0.828 microg/ml and 0.414 microg/ml achieved by colorimetric and chemiluminescent ELISAs, respectively. These results, together with the additional advantages of rapidity, lower reagent volumes and moderate operating conditions, have set the grounds for further study in order to adapt this platform for on-site diagnostics of brucellosis disease markers.

摘要

免疫传感器是临床和兽医诊断中强有力的分析工具。这促使我们设计了一种化学发光免疫传感器,旨在使用光纤作为换能器来识别抗布鲁氏菌抗体。为了开发最佳的换能器,实现最佳的化学修饰,从而允许蛋白质受体的最佳共价结合,我们研究了几种清洗策略和硅烷偶联剂。使用布鲁氏菌杀死的生物体作为模型受体,与传统的比色法和化学发光 ELISA 相比,在悬浮液中定量抗布鲁氏菌 IgG 抗体。选择硅烷-二苯甲酮衍生物作为性能最佳的硅烷偶联剂:光纤免疫传感器(OFIS)的检测限最低,为 0.207μg/ml,比比色法和化学发光 ELISA 分别达到的 0.828μg/ml 和 0.414μg/ml 更低。这些结果,加上快速、试剂体积更小和中等操作条件等额外优势,为进一步研究奠定了基础,以便将该平台适用于布鲁氏菌病标志物的现场诊断。

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