Screening Aptamers that Are Specific for Beclomethasone and the Development of Quantum Dot-Based Assay

被引:1
|
作者
Fan, Hongli [1 ]
Liu, Yaxiong [2 ]
Dong, Jiamei [1 ]
Luo, Zhuoya [2 ]
机构
[1] Guangzhou Univ Chinese Med, Inst Math Engn, Guangzhou 510006, Peoples R China
[2] Guangdong Inst Drug Control, NMPA Key Lab Rapid Testing Technol Drugs, Guangzhou 510663, Peoples R China
关键词
Beclomethasone; Systematic evolution of ligands by exponential enrichment; SELEX; Aptamers; Quantum dot-based assay; IONIZATION MASS-SPECTROMETRY; IN-VITRO SELECTION; DNA APTAMERS; RNA APTAMERS; BIOSENSOR; BINDING; PROTEIN; CORTICOSTEROIDS; RECOGNITION; MECHANISM;
D O I
10.1007/s12010-021-03585-x
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
We developed an aptamer that was specific for beclomethasone (BEC) via systematic evolution of ligands by exponential enrichment (SELEX). Development was monitored by real-time quantitative PCR (Q-PCR) and the enriched library was sequenced by high-throughput sequencing. Forty-seven aptamer candidates were obtained; of these, BEC-6 showed the highest affinity (K-d = 0.15 +/- 0.02 mu M) and did not cross-react with other BEC analogs. We also developed a quantum dot-based assay (QDA) for the detection of BEC that was based upon a quantum dot (QD) composite probe. Under optimized reaction conditions, the linear range of this method for BEC was 0.1 to 10 mu M with a low detection limit (LOD) of 0.1 mu M. Subsequently, the method was used to detect BEC in Traditional Chinese Medicine (TCM) with a mean recovery of 81.72-91.84%. This is the first report to describe the development of an aptamer against BEC; BEC-6 can also be engineered into QDA for the detection of BEC.
引用
收藏
页码:3139 / 3150
页数:12
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