Multiplexed detection of small analytes by structure-switching aptamer-based capillary electrophoresis.

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  • Additional Information
    • Source:
      Publisher: American Chemical Society Country of Publication: United States NLM ID: 0370536 Publication Model: Print Cited Medium: Internet ISSN: 1520-6882 (Electronic) Linking ISSN: 00032700 NLM ISO Abbreviation: Anal Chem Subsets: MEDLINE
    • Publication Information:
      Original Publication: Washington, American Chemical Society.
    • Subject Terms:
      Aptamers, Nucleotide/*chemistry ; Aptamers, Nucleotide/*metabolism ; Biosensing Techniques/*methods ; Electrophoresis, Capillary/*methods; Adenosine Monophosphate/analysis ; Adenosine Monophosphate/metabolism ; Base Sequence ; Chromatography, Micellar Electrokinetic Capillary ; Fluorescein/metabolism ; Fluorescent Dyes/metabolism ; Lasers ; Molecular Weight ; Motion ; Nucleic Acid Hybridization ; Spectrometry, Fluorescence ; Time Factors
    • Abstract:
      Affinity probe capillary electrophoresis (APCE) assays, combining the separation power of CE with the specificity of interactions occurring between a target and a molecular recognition element (MRE), have become important analytical tools in many application fields. In this report, a rationalized strategy, derived from the structure-switching aptamer concept, is described for the design of a novel APCE mode dedicated to small molecule detection. Two assay configurations were reported. The first one, developed for the single-analyte determination, was based on the use of a cholesteryl-tagged aptamer (Chol-Apt) as the MRE and its fluorescein-labeled complementary strand (CS*) as the tracer (laser-induced fluorescence detection). Under micellar electrokinetic chromatography (MEKC) conditions, free CS* and the hybrid formed with Chol-Apt (duplex*) were efficiently separated (and then quantified) through the specific shift of the electrophoretic mobility of the cholesteryl-tagged species in the presence of a neutral micellar phase. When the target was introduced into the preincubated sample, the hybridized form was destabilized, resulting in a decrease in the duplex* peak area and a concomitant increase in the free CS* peak area. The second format, especially designed for multianalyte sensing, employed dually cholesteryl- and fluorescein-labeled complementary strands (Chol-CS*) of different lengths and unmodified aptamers (Apt). The size-dependent electrophoretic separation of different Chol-CS* forms from each other and from their corresponding duplexes* was also accomplished under MEKC conditions. The simultaneous detection of multiple analytes in a single capillary was performed by monitoring accurately each target-induced duplex-to-complex change. This method could expand significantly the potential of small solute APCE analysis in terms of simplicity, adaptability, generalizability, and high-throughput analysis capability.
    • Accession Number:
      0 (Aptamers, Nucleotide)
      0 (Fluorescent Dyes)
      415SHH325A (Adenosine Monophosphate)
      TPY09G7XIR (Fluorescein)
    • Publication Date:
      Date Created: 20100508 Date Completed: 20100902 Latest Revision: 20151119
    • Publication Date:
      20221213
    • Accession Number:
      10.1021/ac100755q
    • Accession Number:
      20446673