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02897nam a2200313Ia 4500 |
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10-1093-clinchem-hvab229 |
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220425s2022 CNT 000 0 und d |
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|a 00099147 (ISSN)
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|a Multiplexed Assay to Quantify the PP-Fold Family of Peptides in Human Plasma Using Microflow Liquid Chromatography-Tandem Mass Spectrometry
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|b Oxford University Press
|c 2022
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|a 11
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856 |
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|z View Fulltext in Publisher
|u https://doi.org/10.1093/clinchem/hvab229
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|a 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.
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|a DPP4
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|a immunoassay
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|a micro-UHPLC-MS/MS
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|a neuropeptide Y (NPY)
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|a pancreatic polypeptide (PP)
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|a peptide YY (PYY)
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|a Bally, L.
|e author
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|a Eugster, P.J.
|e author
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|a Grouzmann, E.
|e author
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|a Herzig, D.
|e author
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|a Kuenzli, C.
|e author
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|a Nakas, C.T.
|e author
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|a Reverter-Branchat, G.
|e author
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|a Rindlisbacher, B.
|e author
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|a Stauffer, T.
|e author
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773 |
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|t Clinical Chemistry
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