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

  • Ge YANG
  • , Shi Miao HAN
  • , Li Ping ZHAO
  • , Chao ZHU
  • , Yuan Yu HUANG
  • , Feng QU*
  • *此作品的通讯作者

科研成果: 期刊稿件文章同行评审

摘要

A DNA aptamer screening method for human immunoglobulin G (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 the samples 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 the 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. Aptamers (Seq Fc1–3) were obtained under optimized incubation conditions (10 mmol/L Na2HPO4-KH2PO4 (pH 7.17), 0.05 mmol/L Mg2+, incubated at 37 °C for 25 min) by three rounds of CE-SELEX, with KD values of 0.071–0.321 μmol/L, and their affinity and specificity were verified by AuNPs colorimetry assay.

源语言英语
页(从-至)601-607
页数7
期刊Chinese Journal of Analytical Chemistry
48
5
DOI
出版状态已出版 - 5月 2020

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