A malachite green light-up aptasensor for the detection of theophylline

TALANTA

卷:232

Article

Sett, Arghya¹  Zara, Lorena^1,2  Dausse, Eric¹  Toulme, Jean-Jacques^1,2 

[1] Univ Bordeaux, U1212, UMR5320, ARNA,CNRS,Inserm, F-33000 Bordeaux, France

[2] Novaptech, F-33600 Pessac, France

关键词: Aptamer;    Malachite green;    Fluorescence;    RNA hairpin;    Kissing complex;    Biosensor;   

DOI  :  10.1016/j.talanta.2021.122417

来源: Elsevier

【 摘 要 】

Biosensors are of interest for the quantitative detection of small molecules (metabolites, drugs and contaminants for instance). To this end, fluorescence is a widely used technique that is easily associated to aptamers. Light-up aptamers constitute a particular class of oligonucleotides that, specifically induce fluorescence emission when binding to cognate fluorogenic ligands such as malachite green (MG). We engineered a dual aptasensor for theophylline (Th) based on the combination of switching hairpin aptamers specific for MG on the one hand and for Th on the other hand, hence their names: malaswitch (Msw) and theoswitch (Thsw). The two aptaswitches form a loop-loop or kissing Msw-Thsw complex only in the presence of theophylline, allowing binding of MG, subsequently generating a fluorescent signal. The combination of the best Msw and Thsw variants, MswG12 and Thsw19.1, results in a 20-fold fluorescence enhancement of MG at saturating theophylline concentration. This aptasensor discriminates between theophylline and its analogues caffeine and theobromine. Kissing aptaswitches derived from light-up aptamers constitute a novel sensing device.

【 授权许可】

Free