2026-04-17T14:07:47Zhttps://recercat.cat/oai/request

oai:recercat.cat:2445/1231902025-12-04T19:37:27Zcom_2072_1057col_2072_478813col_2072_478917

Analysis of circulating microRNAs and their post-transcriptional modifications in cancer serum by on-line solid-phase extraction-capillary electrophoresis-mass spectrometry Peró Gascón, Roger Sanz Nebot, María Victoria Berezovski, Maxim V. Benavente Moreno, Fernando J. (Julián) Electroforesi capil·lar Espectrometria de masses Micro RNAs Capillary electrophoresis Mass spectrometry MicroRNAs In this paper, an on-line solid-phase extraction capillary electrophoresis-mass spectrometry (SPE-CE-MS) method is described for the purification, preconcentration, separation, and characterization of endogenous microRNA (miRNA) and their post-transcriptional modifications in serum. First, analysis by CE-MS was optimized using a standard mixture of hsa-miR-21-5p (miR-21-5p) and hsa-let-7g-5p (let-7g-5p). For SPE-CE-MS, a commercial silicon carbide (SiC) resin was used to prepare the microcartridges. Under the optimized conditions with standards, the microcartridge lifetime (>25 analyses) and repeatability (2.8% RSD for the migration times; 4.4 and 6.4% RSD for the miR-21-5p and let-7g-5p peak areas, respectively) were good, the method was linear between 25 and 100 nmol·L-1, and the limit of detection (LOD) was around 10 nmol·L-1 (50 times lower than by CE-MS). In order to analyze human serum samples, an off-line sample pretreatment based on phenol/chloroform/isoamyl alcohol (PCA) extraction was necessary prior to SPE-CE-MS. The potential of the SPE-CE-MS method to screen for B-cell chronic lymphocytic leukemia (CLL) was demonstrated by an analysis of serum samples from healthy controls and patients. MicroRNAs, specifically miR-21-5p and a 23 nucleotide long 5'-phosphorylated miRNA with 3'-uridylation (iso-miR-16-5p), were only detected in the CLL patients. 2018-06-21T15:03:28Z 2019-05-08T05:10:13Z 2018-05-07 2018-06-21T15:03:28Z info:eu-repo/semantics/article info:eu-repo/semantics/acceptedVersion 0003-2700 https://hdl.handle.net/2445/123190 680822 29730931 eng Versió postprint del document publicat a: https://doi.org/10.1021/acs.analchem.8b00405 Analytical Chemistry, 2018, vol. 90, p. 6618-6625 https://doi.org/10.1021/acs.analchem.8b00405 (c) American Chemical Society , 2018 info:eu-repo/semantics/openAccess 8 p. application/pdf American Chemical Society Articles publicats en revistes (Enginyeria Química i Química Analítica)