Three-step evolutionary enhanced capillary electrophoresis-SELEX for aptamer selection of exosome vesicles

Liping Zhao; Ge Yang; Chao Zhu; Linsen Li; Yi Zhao; Yunxia Luan; Feng Qu

doi:10.1016/j.talanta.2023.125203

Three-step evolutionary enhanced capillary electrophoresis-SELEX for aptamer selection of exosome vesicles

Talanta. 2024 Jan 15:267:125203. doi: 10.1016/j.talanta.2023.125203. Epub 2023 Sep 14.

Authors

Liping Zhao¹, Ge Yang², Chao Zhu³, Linsen Li⁴, Yi Zhao⁴, Yunxia Luan⁵, Feng Qu⁶

Affiliations

¹ Institute of Quality Standard and Testing Technology of BAAFS, Agricultural Product Quality and Safety Risk Assessment Laboratory of the Department of Agriculture, Beijing 100097, China; Key Laboratory of Molecular Medicine and Biotherapy, School of Life Science, Beijing Institute of Technology, 5 South Zhongguancun Street, Beijing 100081, China.
² CAMS Key Laboratory of Antiviral Drug Research, Beijing Key Laboratory of Antimicrobial Agents, NHC Key Laboratory of Biotechnology of Antibiotics, Institute of Medicinal Biotechnology, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing 100050, China.
³ Institute of Quality Standard and Testing Technology for Agro-products, Shandong Academy of Agricultural Sciences, Jinan 250100, China.
⁴ Key Laboratory of Molecular Medicine and Biotherapy, School of Life Science, Beijing Institute of Technology, 5 South Zhongguancun Street, Beijing 100081, China.
⁵ Institute of Quality Standard and Testing Technology of BAAFS, Agricultural Product Quality and Safety Risk Assessment Laboratory of the Department of Agriculture, Beijing 100097, China.
⁶ Key Laboratory of Molecular Medicine and Biotherapy, School of Life Science, Beijing Institute of Technology, 5 South Zhongguancun Street, Beijing 100081, China. Electronic address: Qufengqu@bit.edu.cn.

PMID: 37748272
DOI: 10.1016/j.talanta.2023.125203

Abstract

As various disease biomarkers, drugs/biomolecules carriers and physiological function regulator, exosomes play significant roles in various physiological and pathological process. Thus, the purification and detection of some exosome are of great significance, which make it necessary for the development of the recognition elements. Aptamers with high affinity and specificity exhibited well in the above areas. In this work, a new three-step evolutionary enhanced strategy based on capillary electrophoresis (CE)-SELEX was proposed for effective aptamer selection of exosome vesicles for the first time. Natural killer cells (NK) exosome was used as the model target. And the aptamer could be obtained in five round CE selection within three steps evolution including precise nucleic acid sequence convergence (with NK exosome specific proteins as targets in round 1st and 3rd), affinity evolution (with NK exosome as target in round 2nd and 4th) and specific evolution (negative selection in round 5th). Arising from the compatibility of CE-SELEX for exosome vesicles aptamer selection, and the proposed strategy, aptamer against NK exosome had been selected with good affinity (K_D value of 27.6 nM) and excellent specificity. The as proposed CE-SELEX strategy paves a novel way for recognition element selection of exosome vesicles.

Keywords: Aptamer; CE-SELEX; NK exosome; Selection strategy.

MeSH terms

Aptamers, Nucleotide* / genetics
Aptamers, Nucleotide* / metabolism
Drug Carriers
Electrophoresis, Capillary / methods
Exosomes* / metabolism
SELEX Aptamer Technique / methods

Substances

Aptamers, Nucleotide
Drug Carriers