Capillary Electrophoresis–SELEX Selection of Catalytic DNA Aptamers for a Small-Molecule Porphyrin Target
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 tar...
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Veröffentlicht in: | Analytical chemistry (Washington) 2013-02, Vol.85 (3), p.1525-1530 |
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Format: | Artikel |
Sprache: | eng |
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Zusammenfassung: | 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. |
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ISSN: | 0003-2700 1520-6882 |
DOI: | 10.1021/ac302721j |