Citation: YANG Ge, ZHU Chao, LIU Xiao-Hui, WANG Yong, QU Feng. Screening of Clenbuterol Hydrochloride Aptamers Based on Capillary Electrophoresis[J]. Chinese Journal of Analytical Chemistry, ;2018, 46(10): 1595-1603. doi: 10.11895/j.issn.0253-3820.181347 shu

Screening of Clenbuterol Hydrochloride Aptamers Based on Capillary Electrophoresis

School of Life Science, Beijing Institute of Technology, Beijing 100081, China

Received Date: 24 May 2018
Revised Date: 29 July 2018

Fund Project: This work was supported by the National Natural Science Foundation of China (No. 21675012)

Capillary electrophoresis based systematic evolution of ligands via exponential enrichment (CE-SELEX) was reported as a homogeneous efficient method for high-affinity selection of aptamer, with several merits involving screening in free solution without nonspecific binding, capable of high-efficient separation, low-sample consumption, and saving money. There are few studies regarding the aptamer selection against small molecule using CE-SELEX, resulting from the aspects of less binding sites and the negligible variety of its complex with nucleic acid in the electrophoretic mobility. In this study, the aptamer selection was performed towards a small molecule target of clenbuterol hydrochloride (Cle) by CE-SELEX. In brief, incubated Cle and an 80 nt ssDNA library were firstly incubated, and CZE-UV approach was used to separate complex and random ssDNA. The complex was then collected into a vial followed by PCR amplification. Through three round selections, the third pool was selected to clone and ten sequences were finally obtained. The dissociation constants (K_d) of three potential candidates (Apt 4, Apt 7 and Apt 12) were determined by CE-LIF, and showed high affinities of 9.315×10^-7 mol/L, 1.040×10^-6 mol/L and 1.143×10^-5 mol/L, respectively. The m-Fold software analysis showed that the above three sequences could form stem-loop structure, and Apt 4 gave the lowest free energy and the most stable structure. Using salbutamol as a control, three selected aptamers were verified with high specificity.
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