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基于石墨烯的化学发光共振能量转移用于人绒毛膜促性腺激素测定的简单灵敏免疫分析。

A simple and sensitive immunoassay for the determination of human chorionic gonadotropin by graphene-based chemiluminescence resonance energy transfer.

机构信息

Key Laboratory of Environment and Health, Ministry of Education & Ministry of Environmental Protection, Tongji Medical College, Huazhong University of Science and Technology, #13 Hangkong Road, Wuhan, Hubei 430030, China; State Key Laboratory of Environmental Health (Incubating), Tongji Medical College, Huazhong University of Science and Technology, #13 Hangkong Road, Wuhan, Hubei 430030, China.

Key Laboratory of Environment and Health, Ministry of Education & Ministry of Environmental Protection, Tongji Medical College, Huazhong University of Science and Technology, #13 Hangkong Road, Wuhan, Hubei 430030, China; State Key Laboratory of Environmental Health (Incubating), Tongji Medical College, Huazhong University of Science and Technology, #13 Hangkong Road, Wuhan, Hubei 430030, China.

出版信息

Biosens Bioelectron. 2014 Apr 15;54:72-7. doi: 10.1016/j.bios.2013.10.033. Epub 2013 Oct 31.

Abstract

In this study, we report a strategy of chemiluminescence resonance energy transfer (CRET) using graphene as an efficient long-range energy acceptor. Magnetic nanoparticles were also used in CRET for simple magnetic separation and immobilization of horseradish peroxidase (HRP)-labeled anti-HCG antibody. In the design of CRET system, the sandwich-type immunocomplex was formed between human chorionic gonadotropin (HCG, antigen) and two different antibodies bridged the magnetic nanoparticles and graphene (acceptors), which led to the occurrence of CRET from chemiluminescence light source to graphene. After optimizing the experimental conditions, the quenching of chemiluminescence signal depended linearly on the concentration of HCG in the range of 0.1 mIU mL(-1)-10 mIU mL(-1) and the detection limit was 0.06 mIU mL(-1). The proposed method was successfully applied for the determination of HCG levels in saliva and serum samples, and the results were in good agreement with the plate ELISA with colorimetric detection. It could also be developed for detection of other antigen-antibody immune complexes by using the corresponding antigens and respective antibodies.

摘要

在本研究中,我们报告了一种使用石墨烯作为高效远程能量受体的化学发光共振能量转移 (CRET) 策略。磁纳米粒子也被用于 CRET 中,用于简单的磁分离和辣根过氧化物酶 (HRP) 标记的抗人绒毛膜促性腺激素 (HCG) 抗体的固定。在 CRET 系统的设计中,夹心型免疫复合物在人绒毛膜促性腺激素 (HCG,抗原) 和两种不同的抗体之间形成,该抗体桥接了磁性纳米粒子和石墨烯 (受体),从而导致化学发光光源到石墨烯的 CRET 发生。在优化实验条件后,化学发光信号的猝灭与 HCG 在 0.1 mIU mL(-1)-10 mIU mL(-1) 范围内的浓度呈线性关系,检测限为 0.06 mIU mL(-1)。该方法成功地应用于唾液和血清样品中 HCG 水平的测定,与比色检测的平板 ELISA 结果具有良好的一致性。通过使用相应的抗原和各自的抗体,也可以开发用于检测其他抗原-抗体免疫复合物的方法。

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