Capillary Electrophoresis-SELEX Selection of Catalytic DNA Aptamers for a Small-Molecule Porphyrin Target

被引：117
|
作者：
Yang, Jing [1 ]
Bowser, Michael T. [1 ]
机构：
[1] Univ Minnesota, Dept Chem, Minneapolis, MN 55455 USA
来源：
ANALYTICAL CHEMISTRY | 2013年 / 85卷 / 03期
基金：
美国国家卫生研究院;
关键词：
IN-VITRO SELECTION; RNA LIGANDS; EQUILIBRIUM MIXTURES; AFFINITY; METALATION; BINDING; OLIGONUCLEOTIDE; INHIBIT; SEPARATION; EVOLUTION;
D O I：
10.1021/ac302721j
中图分类号：
O65 [分析化学];
学科分类号：
070302 ; 081704 ;
摘要：
Capillary electrophoresis-systematic evolution of ligands by exponential enrichment (CE-SELEX) has previously been used to select aptamers for large-molecule targets such as proteins, lipopolysaccharides, and peptides. For the first time, we have performed CE-SELEX selection for a small-molecule target, N-methyl mesoporphyrin (NMM), with a molecular weight of only 580 g/mol. DNA aptamers with high-nanomolar to low-micromolar dissociation constants were achieved after only three rounds of selection. This corresponds to an >50-fold improvement in affinity over the random library. Two out of eight randomly chosen aptamers were found to catalyze the metal insertion reaction of mesoporphyrin with 1.7- and 2.0-fold rate enhancements, respectively.
引用
收藏
页码：1525 / 1530
页数：6
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