Fan Hongli, Liu Yaxiong, Dong Jiamei, Luo Zhuoya
Institute of Mathematical Engineering, Guangzhou University of Chinese Medicine, Guangzhou, 510006, China.
NMPA Key Laboratory for Rapid Testing Technology of Drugs, Guangdong Institute for Drug Control, Guangzhou, 510663, China.
Appl Biochem Biotechnol. 2021 Oct;193(10):3139-3150. doi: 10.1007/s12010-021-03585-x. Epub 2021 Jun 3.
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 = 0.15 ± 0.02 μM) 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 μM with a low detection limit (LOD) of 0.1 μM. 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.
我们通过指数富集配体系统进化技术（SELEX）开发了一种对倍氯米松（BEC）具有特异性的适配体。通过实时定量聚合酶链反应（Q-PCR）监测开发过程，并通过高通量测序对富集文库进行测序。获得了47个候选适配体；其中，BEC-6表现出最高的亲和力（K = 0.15±0.02 μM），且不与其他BEC类似物发生交叉反应。我们还开发了一种基于量子点的检测方法（QDA）用于检测BEC，该方法基于量子点（QD）复合探针。在优化的反应条件下，该方法对BEC的线性范围为0.1至10 μM，检测限低至0.1 μM。随后，该方法用于检测中药中的BEC，平均回收率为81.72-91.84%。这是第一份描述针对BEC的适配体开发的报告；BEC-6也可用于构建QDA以检测BEC。
