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Rapid selection of aptamers based on protein microarray

Author:
Yu, Fang, Li, Hui, Sun, Wei, Xu, Danke, He, Fuchu
Source:
RSC advances 2019 v.9 no.17 pp. 9762-9768
ISSN:
2046-2069
Subject:
beta-lactoglobulin, bovine serum albumin, casein, dissociation, fluorescence, lactalbumin, lactoferrin, oligonucleotides, protein microarrays, scanners, screening, single-stranded DNA, systematic evolution of ligands by exponential enrichment
Abstract:
We report a novel method for the efficient screening of aptamers from a complex ssDNA library based on a microarray chip, which was named Microarray-SELEX. In this method, the target protein (lactoferrin) and negative proteins (α-lactalbumin, β-lactoglobulin, bovine serum albumin, and casein) were each dotted and immobilized on a slide to form a protein microarray. Moreover, the library was added to this chip to interact with negative proteins, then with the target protein. The process of SELEX could be monitored on-line using a fluorescent microarray scanner and the whole process was performed in only six rounds. Finally, five aptamers (YFL-1, YFL-4, YFL-5, YFL-6 and YFL-7) were obtained, which showed good specificity towards lactoferrin in the presence of negative proteins. The equilibrium dissociation constants (Kd) of the aptamers were in the nanomolar range. Briefly, Microarray-SELEX is a rapid, easy, sensitive and efficient method for screening aptamers.
Agid:
6354354