Citation: LI Lin-Sen, ZHU Chao, ZHAO Yi, YANG Ge, QU Feng. Investigation of Library Input on Aptamers Selection Efficiency Using Capillary Electrophoresis[J]. Chinese Journal of Analytical Chemistry, ;2020, 48(5): 615-622. doi: 10.19756/j.issn.0253-3820.191693 shu

Investigation of Library Input on Aptamers Selection Efficiency Using Capillary Electrophoresis

School of Life Science, Beijing Institute of Technology, Key Laboratory of Molecular Medicine and Biological Diagnosis, Ministry of Industry and Information Technology, Beijing 100081, China

Received Date: 25 November 2019
Revised Date: 20 January 2020

Fund Project: This work was supported by the National Natural Science Foundation of China (Nos. 21675012, 21874010, 21827810).

Capillary electrophoresis (CE) is a high-efficientcy method for aptamer selection, which exhibits many merits of high resolution, rapid separation, little sample consumption, and no immobilization. However, the influence of CE injection input on aptamer systematic evolution of ligands by exponential enrichment (SELEX) is controversial because the low injection volume may result in an insufficient capacity of the oligonucleotides library. In this research, Neuron-specific enolase (NSE) was used as a model protein to investigate the effects of library input capacity on aptamer selection for clarifying the validity of CE-SELEX. Four different concentrations of initial libraries (0.1, 1, 10 and 100 μmol/L) were utilized into aptamer selection. The affinity of the initial library and sub-libraries after three rounds of selection were evaluated. The results showed that there was no significant difference in the affinity of four different sub-libraries, and the aptamers with micromolar K_D were all obtained after two rounds of selection. In addition, the sequences obtained via high concentration ssDNA library screening were more diverse, but the affinity of them was not enriched. The NSE aptamer Seq qN-01 obtained by two rounds of selection had a dissociation constant (K_D) of (4.72±0.15) μmol/L, and also presented a good specificity. In conclusion, library input capacity could increase the sequence diversity but had no considerable effects on the efficiency of aptamer selection.
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