title: Efficient selection of glycoprotein-binding DNA aptamers via boronate affinity monolithic capillary.
abstract: Systematic evolution of ligands by exponential enrichment (SELEX) is the workhorse method for selecting aptamers that are capable of binding target molecules from a random oligonucleic acid library. However, conventional SELEX methods are associated with apparent drawbacks including labor-intensive, time-consuming, large reagent consumption and strong nonspecific binding with separation media. Herein, we report a boronate affinity monolithic capillary-based SELEX approach for rapid selection of high-specificity glycoprotein-binding DNA aptamers. Boronate affinity monolithic capillary is an advanced functional material appeared in recent years, which allows for facile capture/release of glycoproteins in a pH-switchable fashion. By using boronate affinity monolithic capillary as a platform for target immobilization and aptamer isolation, the proposed SELEX method allowed for efficient selection of glycoprotein-binding aptamers by 6 rounds and the dissociation constants were at 10(-8) M level. Because of the employment of boronate affinity monolithic capillary, the new SELEX approach overcame the above-mentioned drawbacks and provided several significant advantages, including rapid selection speed (only 2 days were needed), high specificity toward the target molecules, and minute reagent consumption.
authors.affiliation: State Key Laboratory of Analytical Chemistry for Life Science, School of Chemistry and Chemical Engineering, Nanjing University, Nanjing 210093, China.
authors.forename: Hongyuan
authors.initials: H
authors.lastname: Nie
authors.forename: Yang
authors.initials: Y
authors.lastname: Chen
authors.forename: Chenchen
authors.initials: C
authors.lastname: Lü
authors.forename: Zhen
authors.initials: Z
authors.lastname: Liu
mesh_terms: D052157:Aptamers, Nucleotide / Q000032:analysis / Q000235:genetics / Q000378:metabolism*; D001483:Base Sequence; D001897:Boronic Acids / Q000737:chemistry / Q000378:metabolism*; D019075:Electrophoresis, Capillary / Q000379:methods; D015723:Gene Library; D006023:Glycoproteins / Q000032:analysis / Q000235:genetics / Q000378:metabolism*; D008969:Molecular Sequence Data; D052156:SELEX Aptamer Technique / Q000379:methods*
chemical_list: D052157:Aptamers, Nucleotide; D001897:Boronic Acids; D006023:Glycoproteins