Selection and characterization of DNA aptamers for highly selective recognition of the major allergen of olive pollen Ole e 1

被引:3
|
作者
Vergara-Barberan, Maria [1 ]
Lerma-Garcia, Maria Jesus [1 ]
Simo-Alfonso, Ernesto F. [1 ]
Garcia-Hernandez, Marta [2 ]
Martin, M. Elena [2 ]
Garcia-Sacristan, Ana [3 ]
Gonzalez, Victor M. [2 ,3 ]
Herrero-Martinez, Jose Manuel [1 ]
机构
[1] Univ Valencia, Dept Analyt Chem, C Doctor Moliner 50, E-46100 Valencia, Spain
[2] Hosp Univ Ramon y Cajal IRYCIS, Grp Aptameros, Dept Bioquim Invest, Carretera Colmenar Viejo Km 9-100, Madrid 28034, Spain
[3] Aptus Biotech SL, Av Cardenal Herrera Oria 298, Madrid 28035, Spain
关键词
Allergen; Olive pollen; Aptamer; Enzyme-linked oligonucleotide assay; Affinity-based monolith; Ole e 1; AMINO-ACID-SEQUENCE; IN-VITRO SELECTION; CHROMATOGRAPHY; BIOMOLECULES; SEPARATION; MONOLITH; SERVER; ASSAY;
D O I
10.1016/j.aca.2021.339334
中图分类号
O65 [分析化学];
学科分类号
070302 ; 081704 ;
摘要
In this study, single-stranded DNA aptamers with binding affinity to Ole e 1, the major allergen of olive pollen, were selected using systematic evolution of ligands by exponential enrichment (SELEX) method. Binding of the aptamers was firstly established by enzyme-linked oligonucleotide assay (ELONA) and aptaprecipitation assays. Additionally, aptamer-modified monolithic capillary chromatography was used in order to evaluate the recognition of this allergenic protein against other non-target proteins. The results indicated that AptOle1#6 was the aptamer that provided the highest affinity for Ole e 1. The selected aptamer showed good selective recognition of this protein, being not able to retain other nontarget proteins (HSA, cyt c, and other pollen protein such as Ole e 9). The feasibility of the affinity monolithic column was demonstrated by selective recognition of Ole e 1 in an allergy skin test. The stability and reproducibility of this monolithic column was suitable, with relative standard deviations (RSDs) in retention times and peak area values of 7.8 and 9.3%, respectively (column-to-column reproducibility). This is the first study that describes the design of an efficient DNA aptamer for this relevant allergen. (c) 2021 The Authors. Published by Elsevier B.V. This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
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