Homogenous FRET-based fluorescent immunoassay for deoxynivalenol detection by controlling the distance of donor-acceptor couple

被引:35
作者
Goryacheva, Olga A. [1 ,2 ]
Beloglazova, Natalia, V [1 ,3 ]
Goryacheva, Irina Yu [2 ]
De Saeger, Sarah [1 ]
机构
[1] Univ Ghent, Fac Pharmaceut Sci, Ctr Excellence Mycotoxicol & Publ Hlth, Ottergemsesteenweg 460, B-9000 Ghent, Belgium
[2] Saratov NG Chernyshevskii State Univ, Chem Inst, Dept Gen & Inorgan Chem, Astrakhanskaya 83, Saratov 410012, Russia
[3] South Ural State Univ, Nanotechnol Educ & Res Ctr, Prospect Lenina 76, Chelyabinsk 485400, Russia
基金
俄罗斯科学基金会;
关键词
Forster resonance energy transfer; Immunoassay; Mycotoxins; Quantum dots; Gold nanoparticles; ENERGY-TRANSFER FRET; QUANTUM-DOTS; SENSITIVE DETECTION; GOLD NANOPARTICLES; ORIENTATION; DIAGNOSTICS; MYCOTOXINS; SIZE;
D O I
10.1016/j.talanta.2020.121973
中图分类号
O65 [分析化学];
学科分类号
070302 ; 081704 ;
摘要
Semiconductor quantum dots (QDs) are one of the most popular luminescent labels that are widely used in food and medical analysis. Their unique optical properties establish QDs as excellent tools for highly sensitive biosensors based on Forster resonance energy transfer (FRET). To provide a convenient analytical system with long-term optical stability, a FRET pair consisting of QDs as energy donor and gold nanoparticles (GNs) as energy acceptor was developed. Careful selection of donor and acceptor properties allowed to achieve a large Forster distance of 12.9 nm and to use full-size specific antibody. As the immunoreagents pair, mycotoxins were bound to proteins and then to GNs, while QDs were conjugated with specific antibodies. FRET was observed as a result of the immunocomplex formation. Contributions of FRET and inner filter effect on the quenching were evaluated separately. The quenching effect in the donor-acceptor pair was compared for proteins with different sizes. The developed homogeneous FRET-based immunoassay for the detection of deoxynivalenol (DON) is an example of a fast method for high-throughput control of mycotoxins. The quenching effect of FRET was observed with a limit of detection of 28 mu g kg(-1) of DON in spiked wheat samples.
引用
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页数:8
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