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基于负载金纳米粒子酶的简单灵敏流动注射化学发光免疫分析法测定氯霉素。

A simple and sensitive flow injection chemiluminescence immunoassay for chloramphenicol based on gold nanoparticle-loaded enzyme.

机构信息

The Key Laboratory of Health Chemistry & Molecular Diagnosis of Suzhou, College of Chemistry, Chemical Engineering & Materials Science, Soochow University, Suzhou, China.

出版信息

Luminescence. 2020 Sep;35(6):877-884. doi: 10.1002/bio.3795. Epub 2020 Mar 9.

Abstract

A simple and ultrasensitive flow injection chemiluminescence competitive immunoassay based on gold nanoparticle-loaded enzyme for the detection of chloramphenicol (CAP) residues in shrimp and honey has been developed. Due to their good biocompatibility and large specific surface area, carboxylic resin beads can be used as solid phase carriers to immobilize more coating antigens (Ag). In addition, gold nanoparticles could provide an effective matrix for loading more CAP antibody and horseradish peroxidase, which would effectively catalyze the system of luminol-p-iodophenol (PIP)-H O . A competitive immunoassay strategy was used for detection of CAP, in which CAP in the sample would compete with the coating Ag for the limited antibodies, leading to a chemiluminescence (CL) signal decrease with increase in CAP concentration. A wide linear range 0.001-10 ng ml (R = 0.9961) was obtained under optimized conditions, and the detection limit (3σ) was calculated to be 0.33 pg ml . This method was also been successfully applied to determine CAP in shrimp and honey samples. The immunosensor proposed in this study not only has the advantages of high sensitivity, wider linear range, and satisfactory stability, but also expands the application of flow injection CL immunoassay in antibiotic detection.

摘要

基于金纳米粒子负载酶的简单、超灵敏流动注射化学发光竞争免疫分析法,用于检测虾和蜂蜜中的氯霉素(CAP)残留。由于其良好的生物相容性和较大的比表面积,羧酸树脂珠可用作固定相载体来固定更多的包被抗原(Ag)。此外,金纳米粒子可以为负载更多的 CAP 抗体和辣根过氧化物酶提供有效的基质,这将有效地催化鲁米诺-p-碘苯酚(PIP)-H₂O 的体系。采用竞争免疫分析策略检测 CAP,样品中的 CAP 会与包被 Ag 竞争有限的抗体,导致 CAP 浓度增加时化学发光(CL)信号降低。在优化条件下,获得了 0.001-10ngml(R=0.9961)的宽线性范围,检测限(3σ)计算为 0.33pgml。该方法还成功地用于测定虾和蜂蜜样品中的 CAP。本研究中提出的免疫传感器不仅具有灵敏度高、线性范围宽、稳定性好等优点,而且扩展了流动注射 CL 免疫分析在抗生素检测中的应用。

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