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Citation: YANG Ge, HAN Shi-Miao, ZHAO Li-Ping, ZHU Chao, HUANG Yuan-Yu, QU Feng. Screening of Aptamer for Human IgG Fc Fragment by Capillary Electrophoresis-Systematic Evolution of Ligands by Exponential Enrichment[J]. Chinese Journal of Analytical Chemistry, ;2020, 48(5): 601-607. doi: 10.19756/j.issn.0253-3820.191761 shu

Screening of Aptamer for Human IgG Fc Fragment by Capillary Electrophoresis-Systematic Evolution of Ligands by Exponential Enrichment

YANG Ge ^1,2, ,
HAN Shi-Miao ¹ ,
ZHAO Li-Ping ¹ ,
ZHU Chao ¹ ,
HUANG Yuan-Yu ^1,2 ,
QU Feng ¹

1.
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;
2.
Advanced Research Institute of Multidisciplinary Science, Beijing Institute of Technology, Beijing 100081, China

Received Date: 24 December 2019
Revised Date: 15 January 2020

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

A DNA aptamer screening method for human IgG Fc fragments was established based on capillary electrophoresis-systematic evolution of ligands by exponential enrichment (CE-SELEX). The purity, charged properties and non-adsorptivity of IgG, Fc and Fab were analyzed by capillary zone electrophoresis (CZE), and their affinity with ssDNA libraries was compared. The results showed that IgG formed significant complex with the ssDNA libraries, while Fc and Fab fragments exhibited weak affinity with ssDNA libraries. Therefore, the use of the Fab fragment as the reverse screening target had no significant effect on the sequence except for the specific binding of Fc. The aptamer screening strategy for Fc fragments was designed, the aptamers (Seq Fc1-3) were obtained under optimized incubation condition (10 mmol/L Na₂HPO₄-KH₂PO₄ (pH 7.17), 0.05 mmol/L Mg²⁺, incubated at 37℃ for 25 min) by three round of CE-SELEX, K_D was 0.071-0.321 μmol/L, and their affinity and specificity were verified by AuNPs colorimetry.
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