Dipeptidyl peptidase 4 (DPP4) in fecal samples: validation of the extraction methodology and stability in short-term storage conditions.

Autor:	Gomes SF; Department of Biomedicine, Unit of Pharmacology and Therapeutics, Faculty of Medicine, University of Porto, Porto, Portugal.; Department of Public Health and Forensic Sciences and Medical Education, Faculty of Medicine, University of Porto, Porto, Portugal.; Center for Drug Discovery and Innovative Medicines (MedInUP), University of Porto, Porto, Portugal., Melo FJ; Department of Biomedicine, Unit of Pharmacology and Therapeutics, Faculty of Medicine, University of Porto, Porto, Portugal.; Center for Drug Discovery and Innovative Medicines (MedInUP), University of Porto, Porto, Portugal., Silva R; Portuguese Study Group of Inflammatory Bowel Disease (GEDII), Porto, Portugal., Santiago M; Portuguese Study Group of Inflammatory Bowel Disease (GEDII), Porto, Portugal., Estevinho MM; Department of Biomedicine, Unit of Pharmacology and Therapeutics, Faculty of Medicine, University of Porto, Porto, Portugal.; Center for Drug Discovery and Innovative Medicines (MedInUP), University of Porto, Porto, Portugal.; Department of Gastroenterology, Vila Nova de Gaia/Espinho Hospital Center, Vila Nova de Gaia, Portugal., Dias S; Portuguese Study Group of Inflammatory Bowel Disease (GEDII), Porto, Portugal., Dias CC; Department of Community Medicine, Information and Health Decision Sciences (MEDCIDS), Faculty of Medicine, University of Porto, Porto, Portugal.; Center for Health Technology and Services Research (CINTESIS), Porto, Portugal., Magro F; Department of Biomedicine, Unit of Pharmacology and Therapeutics, Faculty of Medicine, University of Porto, Porto, Portugal.; Portuguese Study Group of Inflammatory Bowel Disease (GEDII), Porto, Portugal.; RISE - Health Research Network, Porto, Portugal.; Department of Gastroenterology, São João University Hospital Center, Porto, Portugal.; Clinical Pharmacology Unit, São João University Hospital Center, Porto, Portugal.
Jazyk:	angličtina
Zdroj:	Clinical chemistry and laboratory medicine [Clin Chem Lab Med] 2023 Apr 25; Vol. 61 (9), pp. 1636-1642. Date of Electronic Publication: 2023 Apr 25 (Print Publication: 2023).
DOI:	10.1515/cclm-2023-0139
Abstrakt:	Objectives: This study assesses the clinical relevance of dipeptidyl peptidase 4 (DPP4) membrane exopeptidase as a biomarker of inflammatory bowel disease (IBD). A spike-and-recovery approach of DPP4 in fecal samples was used to compare two different methods for protein extraction, followed by a stability assessment. Methods: Fecal samples of healthy volunteers spiked with known concentrations of recombinant DPP4 were processed using a standard manual extraction protocol and the CALEX ® protocol. The two methods were compared by quantification of fecal DPP4 by ELISA, followed by Bland-Altman analysis. For the stability assays DPP4 was extracted from fecal samples and stored under different conditions of temperature and time after collection. Results: In general, the levels of spiked DPP4 in stool samples were lower with the manual protocol than in those obtained with the CALEX ® method; this trend was corroborated by Bland-Altman analysis. Nonetheless, variability was within the acceptable limits for both protocols. In the stability assessment, no statistically significant differences were found between the results obtained under the different storage conditions. Conclusions: Both manual and CALEX ® protocols provided equal extraction ability of DPP4 from stool samples. In addition, DPP4 provided flexibility in terms of sample storage enabling the accurate assessment of samples delivered up to a week before analysis. (© 2023 Walter de Gruyter GmbH, Berlin/Boston.)
Databáze:	MEDLINE
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