Aptamers as ligands in affinity probe capillary electrophoresis.
A DNA aptamer against IgE was labeled with fluorophore and used as a selective fluorescent tag for determining IgE by capillary electrophoresis with laser-induced fluorescence detection (CE-LIF). CE-LIF separations of samples containing fluorescently labeled aptamer and IgE were complete in less than 60 s and revealed two zones, one corresponding to free aptamer and the other to aptamer bound to IgE. The free aptamer peak decreased and bound aptamer peak increased in proportion to the amount of IgE in the sample so that IgE could be detected with a linear dynamic range of 10(5) and a detection limit of 46 pM. The assay was highly selective as aptamer was unaffected by the presence of IgG and IgE did not bind other DNA sequences. IgE was determined in serum samples with similar analytical figures of merit. Similar conditions using a thrombin aptamer allowed detection of thrombin.
German
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Department of Chemistry, University of Florida, Gainesville 32611-7200, USA.
Buchanan
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Kennedy
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D000345:Affinity Labels; D052157:Aptamers, Nucleotide; D004247:DNA / Q000097:blood; D019075:Electrophoresis, Capillary / Q000379:methods*; D019855:Ethylene Glycol; D016650:Fluorescein-5-isothiocyanate; D005456:Fluorescent Dyes; D006801:Humans; D007073:Immunoglobulin E / Q000097:blood; D009841:Oligonucleotides; D013917:Thrombin / Q000032:analysis
D000345:Affinity Labels; D052157:Aptamers, Nucleotide; D005456:Fluorescent Dyes; D009841:Oligonucleotides; C081676:thrombin aptamer; D007073:Immunoglobulin E; D004247:DNA; D013917:Thrombin; D019855:Ethylene Glycol; D016650:Fluorescein-5-isothiocyanate