Highly selective and sensitive detection of glutamate by an electrochemical aptasensor

被引:16
|
作者
Wu, Changtong [1 ,2 ]
Barkova, Daria [3 ]
Komarova, Natalia [3 ]
Offenhaeusser, Andreas [1 ,2 ]
Andrianova, Mariia [3 ]
Hu, Ziheng [1 ]
Kuznetsov, Alexander [3 ]
Mayer, Dirk [1 ]
机构
[1] Forschungszentrum Julich, Inst Biol Informat Proc, IBI 3, D-52425 Julich, Germany
[2] Rhein Westfal TH Aachen, Fac 1, D-52062 Aachen, Germany
[3] Sci Mfg Complex Technol Ctr, 1-7 Shokin Sq, Moscow 124498, Russia
关键词
Aptamer; Capture-SELEX; Glutamic acid; Electrochemical aptamer-based sensor; RNA APTAMERS; LABEL-FREE; IN-VITRO; SENSOR; RECOGNITION; BRAIN; BIOSENSORS; EVOLUTION; DYNAMICS; AFFINITY;
D O I
10.1007/s00216-021-03783-w
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
An electrochemical aptamer-based sensor was developed for glutamate, the major excitatory neurotransmitter in the central nervous system. Determining glutamic acid release and glutamic acid levels is crucial for studying signal transmission and for diagnosing pathological conditions in the brain. Glutamic acid-selective oligonucleotides were isolated from an ssDNA library using the Capture-SELEX protocol in complex medium. The selection permitted the isolation of an aptamer 1d04 with a dissociation constant of 12 mu M. The aptamer sequence was further used in the development of an electrochemical aptamer sensor. For this purpose, a truncated aptamer sequence named glu1 was labelled with a ferrocene redox tag at the 3 '-end and immobilized on a gold electrode surface via Au-thiol bonds. Using 6-mercapto-1-hexanol as the backfill, the sensor performance was characterized by alternating current voltammetry. The glu1 aptasensor showed a limit of detection of 0.0013 pM, a wide detection range between 0.01 pM and 1 nM, and good selectivity for glutamate in tenfold diluted human serum. With this enzyme-free aptasensor, the highly selective and sensitive detection of glutamate was demonstrated, which possesses great potential for implementation in microelectrodes and for in vitro as well as in vivo monitoring of neurotransmitter release.
引用
收藏
页码:1609 / 1622
页数:14
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