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Multiplexed Assay to Quantify the PP-Fold Family of Peptides in Human Plasma Using Microflow Liquid Chromatography-Tandem Mass Spectrometry
dc.creator | Reverter-Branchat G., Eugster P.J., Kuenzli C., Rindlisbacher B., Stauffer T., Nakas C.T., Herzig D., Grouzmann E., Bally L. | en |
dc.date.accessioned | 2023-01-31T09:51:25Z | |
dc.date.available | 2023-01-31T09:51:25Z | |
dc.date.issued | 2022 | |
dc.identifier | 10.1093/clinchem/hvab229 | |
dc.identifier.issn | 00099147 | |
dc.identifier.uri | http://hdl.handle.net/11615/78496 | |
dc.description.abstract | Background: Peptide Tyr-Tyr (PYY1-36), pancreatic polypeptide (PP1-36) and neuropeptide Y (NPY1-36) constitute the PP-fold family of peptides that is involved in metabolic regulation. Very low plasma concentrations and cleavage into active 3-36 fragments challenge bioanalytical assays used for the quantification of these peptides. Methods: We developed a multiplexed isotopic dilution assay to quantify PYY1-36, PP1-36, and NPY1-36 and their dipeptidyl peptidase-4 (DPP4)-derived metabolites PYY3-36, PP3-36 and NPY3-36. All peptides were immunocaptured from plasma using a monoclonal antibody and quantified by micro-ultra-HPLC-MS/MS. Blood samples from healthy volunteers were collected fasting and 30 min after nutrient stimulation. Method comparison was performed with commercial immunoassays. Results: Linearity was shown in the measured intervals (r2 > 0.99). The lower limit of quantification (LLOQ) with a CV at 20% was 1.5 pM for PYY1-36 and PYY3-36, 3.0 pM for PP1-36 and PP3-36, 0.8 pM for NPY1-36 and 0.5 pM for NPY3-36. In all cases, intra- and inter-assay bias and imprecision were <21%. Pre-analytical stability required addition of a protease inhibitor cocktail. Physiological concentrations of PYY3-36, NPY3-36, PP1-36 and PP3-36 were above the LLOQ in 43% to 100% of the samples. PYY1-36 and NPY1-36 were above the LLOQ in 9% and 0% of the samples, respectively. Immunoassays showed higher concentrations of measurands and poor agreement when compared with micro-UHPLC-MS/MS. Conclusions: The assay allowed for specific multiplexed analysis of the PP-fold family of peptides and their DPP4-cleaved fragments in a single sample, thereby offering new perspectives to study the role and therapeutic potential of these essential peptide hormones in health and metabolic disease. © 2022 American Association for Clinical Chemistry 2022. | en |
dc.language.iso | en | en |
dc.source | Clinical Chemistry | en |
dc.source.uri | https://www.scopus.com/inward/record.uri?eid=2-s2.0-85128160715&doi=10.1093%2fclinchem%2fhvab229&partnerID=40&md5=ddec155f0f188d540fa76822973b5279 | |
dc.subject | 1,10 phenanthroline | en |
dc.subject | 4 (2 aminoethyl)benzenesulfonyl fluoride | en |
dc.subject | actinonin | en |
dc.subject | aprotinin | en |
dc.subject | dipeptidyl peptidase IV | en |
dc.subject | edetic acid | en |
dc.subject | fasidotrilat | en |
dc.subject | leupeptin | en |
dc.subject | n [n (3 carboxyoxirane 2 carbonyl)leucyl]agmatine | en |
dc.subject | neuropeptide Y | en |
dc.subject | pancreas polypeptide | en |
dc.subject | pepstatin | en |
dc.subject | peptide | en |
dc.subject | peptide npy3 36 | en |
dc.subject | peptide pp3 36 | en |
dc.subject | peptide YY | en |
dc.subject | peptide YY [3-36] | en |
dc.subject | proteinase inhibitor | en |
dc.subject | unclassified drug | en |
dc.subject | vildagliptin | en |
dc.subject | adult | en |
dc.subject | Article | en |
dc.subject | blood sampling | en |
dc.subject | dilution | en |
dc.subject | female | en |
dc.subject | human | en |
dc.subject | immunoassay | en |
dc.subject | intermethod comparison | en |
dc.subject | limit of quantitation | en |
dc.subject | liquid chromatography-mass spectrometry | en |
dc.subject | male | en |
dc.subject | micro ultra high pressure liquid chromatography tandem mass spectrometry | en |
dc.subject | normal human | en |
dc.subject | ultra performance liquid chromatography | en |
dc.subject | Oxford University Press | en |
dc.title | Multiplexed Assay to Quantify the PP-Fold Family of Peptides in Human Plasma Using Microflow Liquid Chromatography-Tandem Mass Spectrometry | en |
dc.type | journalArticle | en |
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