Quantitative analysis of microRNA in blood serum with protein-facilitated affinity capillary electrophoresis

  1. Get@NRC: Quantitative analysis of microRNA in blood serum with protein-facilitated affinity capillary electrophoresis (Opens in a new window)
DOIResolve DOI: http://doi.org/10.1021/ac2016213
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Journal titleAnalytical Chemistry
Pages61966201; # of pages: 6
SubjectAffinity capillary electrophoresis; Blood samples; Blood serum; Cancer classification; Detection limits; Double-stranded RNA; Magnetic beads; MicroRNAs; Potential sources; Pre-concentration; Selective binding; Single-stranded DNA; Single-stranded dna binding proteins; Blood; Capillary electrophoresis; Diseases; DNA; Polymerase chain reaction; Probes; Proteins; RNA; Tissue; Blood vessels; DNA binding protein; double stranded RNA; fluorescent dye; microRNA; MIRN122 microRNA, human; RNA binding protein; single stranded DNA; article; biosynthesis; blood; capillary electrophoresis; chemical analysis; chemistry; human; limit of detection; metabolism; methodology; nucleic acid hybridization; RNA probe; Chemistry Techniques, Analytical; DNA, Single-Stranded; DNA-Binding Proteins; Electrophoresis, Capillary; Fluorescent Dyes; Humans; Limit of Detection; MicroRNAs; Nucleic Acid Hybridization; RNA Probes; RNA, Double-Stranded; RNA-Binding Proteins
AbstractMicroRNAs (miRNAs) are small (∼22 nt) regulatory RNAs that are frequently deregulated in cancer and have shown promise as tissue- and blood-based biomarkers for cancer classification and prognostication. Here we present a protein-facilitated affinity capillary electrophoresis (ProFACE) assay for rapid quantification of miRNA levels in blood serum using single-stranded DNA binding protein (SSB) and double-stranded RNA binding protein (p19) as separation enhancers. The method utilizes either the selective binding of SSB to a single-stranded DNA/RNA probe or the binding of p19 to miRNA-RNA probe duplex. For the detection of ultralow amounts of miRNA without polymerase chain reaction (PCR) amplification in blood samples we apply off-line preconcentration of synthetic miRNA-122 from serum by p19-coated magnetic beads followed by online sample stacking in the ProFACE assay. The detection limit is 0.5 fM or 30 000 miRNA molecules in 1 mL of serum as a potential source of naïve miRNAs. © 2011 American Chemical Society.
Publication date
AffiliationNational Research Council Canada (NRC-CNRC); NRC Steacie Institute for Molecular Sciences (SIMS-ISSM)
Peer reviewedYes
NPARC number21271260
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Record identifier26b53f0f-0ba1-451b-b358-3ccbc118b3fd
Record created2014-03-24
Record modified2016-05-09
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