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Selection and identification of an ssDNA aptamer to NB4 cell

This study was to find the aptamers with high affinity and specificity binding to acute promyelocytic leukemia (APL) NB4 cell line. These aptamers targeted NB4 cells were selected from a random single‐stranded DNA (ssDNA) library of systematic evolution of ligands by exponential enrichment (CELL‐SEL...

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Autores principales: Zhang, Xian‐hui, Wang, Wei, Chen, Xin
Formato: Online Artículo Texto
Lenguaje:English
Publicado: John Wiley and Sons Inc. 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8059720/
https://www.ncbi.nlm.nih.gov/pubmed/33522630
http://dx.doi.org/10.1002/jcla.23718
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author Zhang, Xian‐hui
Wang, Wei
Chen, Xin
author_facet Zhang, Xian‐hui
Wang, Wei
Chen, Xin
author_sort Zhang, Xian‐hui
collection PubMed
description This study was to find the aptamers with high affinity and specificity binding to acute promyelocytic leukemia (APL) NB4 cell line. These aptamers targeted NB4 cells were selected from a random single‐stranded DNA (ssDNA) library of systematic evolution of ligands by exponential enrichment (CELL‐SELEX). The binding rate of FITC‐ssDNA library and NB4 cells was monitored using flow cytometry and fluorescence microscope. After cloned and sequenced, the structure, specificity, and affinity of these candidate aptamers were further analyzed. After a total of 19 rounds of selection, the ssDNA library was enriched and the BR (19.9%) of the 16th round was 12 times of the first round (1.6%). Three enriched aptamers were obtained from 21 positive clones of the 16th round, and the predicted secondary structures of these aptamers were mainly stem‐loop. The aptamer CX9 had the highest affinity, and the equilibrium dissociation constant (Kd) was 16.2 nM. The fluorescence intensity of CX9 binding to NB4 cells was stronger than HL60 and K562 cells under fluorescence microscopy. The study indicates that aptamer CX9 exhibits high affinity and specificity with NB4 cells and lay a foundation for the rapid diagnostic method to detect APL with fluorescence‐labeled aptamer.
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spelling pubmed-80597202021-04-23 Selection and identification of an ssDNA aptamer to NB4 cell Zhang, Xian‐hui Wang, Wei Chen, Xin J Clin Lab Anal Research Articles This study was to find the aptamers with high affinity and specificity binding to acute promyelocytic leukemia (APL) NB4 cell line. These aptamers targeted NB4 cells were selected from a random single‐stranded DNA (ssDNA) library of systematic evolution of ligands by exponential enrichment (CELL‐SELEX). The binding rate of FITC‐ssDNA library and NB4 cells was monitored using flow cytometry and fluorescence microscope. After cloned and sequenced, the structure, specificity, and affinity of these candidate aptamers were further analyzed. After a total of 19 rounds of selection, the ssDNA library was enriched and the BR (19.9%) of the 16th round was 12 times of the first round (1.6%). Three enriched aptamers were obtained from 21 positive clones of the 16th round, and the predicted secondary structures of these aptamers were mainly stem‐loop. The aptamer CX9 had the highest affinity, and the equilibrium dissociation constant (Kd) was 16.2 nM. The fluorescence intensity of CX9 binding to NB4 cells was stronger than HL60 and K562 cells under fluorescence microscopy. The study indicates that aptamer CX9 exhibits high affinity and specificity with NB4 cells and lay a foundation for the rapid diagnostic method to detect APL with fluorescence‐labeled aptamer. John Wiley and Sons Inc. 2021-02-01 /pmc/articles/PMC8059720/ /pubmed/33522630 http://dx.doi.org/10.1002/jcla.23718 Text en © 2021 The Authors. Journal of Clinical Laboratory Analysis published by Wiley Periodicals LLC https://creativecommons.org/licenses/by-nc-nd/4.0/This is an open access article under the terms of the http://creativecommons.org/licenses/by-nc-nd/4.0/ (https://creativecommons.org/licenses/by-nc-nd/4.0/) License, which permits use and distribution in any medium, provided the original work is properly cited, the use is non‐commercial and no modifications or adaptations are made.
spellingShingle Research Articles
Zhang, Xian‐hui
Wang, Wei
Chen, Xin
Selection and identification of an ssDNA aptamer to NB4 cell
title Selection and identification of an ssDNA aptamer to NB4 cell
title_full Selection and identification of an ssDNA aptamer to NB4 cell
title_fullStr Selection and identification of an ssDNA aptamer to NB4 cell
title_full_unstemmed Selection and identification of an ssDNA aptamer to NB4 cell
title_short Selection and identification of an ssDNA aptamer to NB4 cell
title_sort selection and identification of an ssdna aptamer to nb4 cell
topic Research Articles
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8059720/
https://www.ncbi.nlm.nih.gov/pubmed/33522630
http://dx.doi.org/10.1002/jcla.23718
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