Repository of Research and Investigative Information

Repository of Research and Investigative Information

Ilam University of Medical Sciences

Selection and characterization of single-stranded DNA aptamers against interleukin-5

Mon Oct 3 11:36:52 2022

(2019) Selection and characterization of single-stranded DNA aptamers against interleukin-5. Research in Pharmaceutical Sciences. pp. 515-523. ISSN 1735-5362

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Abstract

Asthma as a chronic inflammatory disorder is associated with many cytokines like interleukin-5 (IL-5) which plays essential role in eosinophil differentiation and maturation. Accordingly, blockage of IL-5 using mepalizumab has been considered as a promising therapeutic approach for asthma. Despite the monocolonal antibody advantages, some restrictions provided an acceptable background for alternative agents like aptamers which could replace with antibodies. In the current study, aptamer isolation against IL-5 molecule was intended, according to the valuable benefits of aptamers over antibodies. HEK-293T/IL-5 cell was constructed to select aptamer using cell-systematic evolution of ligands by exponential enrichment (SELEX) method. Integration of the IL-5 fragment to genome of the HEK-293T was verified by polymerase chain reaction on the genomic DNA of the transfected cells. Moreover, IL-5 protein expression on the cell surface was confirmed using flow cytometry analysis. Then, cell SELEX was carried out in 12 rounds and isolated aptamers were evaluated by flow cytometry analysis. The selected clones were then sequenced and assessed for any possible secondary structure. The results of this study led to the selection of 19 different single-stranded DNA clones after 12 rounds of selection which were clustered to five groups based on common structural motifs. In conclusion, the findings revealed the isolation of IL-5-specific single-stranded DNA aptamers, which can further be substituted with mepolizumab.

Item Type: Article
Creators:
CreatorsEmail
Jamalvandi, M.UNSPECIFIED
Khanahmad, H.UNSPECIFIED
Irani, S.UNSPECIFIED
Bastaminezhad, S.UNSPECIFIED
Keywords: Aptamer Asthma Cell-SELEX HEK-293T Interleukin-5 cell-surface display optimization mepolizumab expression blocking asthma Pharmacology & Pharmacy
Divisions:
Page Range: pp. 515-523
Journal or Publication Title: Research in Pharmaceutical Sciences
Journal Index: ISI
Volume: 14
Number: 6
Identification Number: https://doi.org/10.4103/1735-5362.272560
ISSN: 1735-5362
Depositing User: مهندس مهدی شریفی
URI: http://eprints.medilam.ac.ir/id/eprint/2707

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