Premium
Sample handling for mass spectrometric proteomic investigations of human urine
Author(s) -
Petri Anette Lykke,
Høgdall Claus,
Christensen Ib Jarle,
Simonsen Anja Hviid,
T'Jampens Davy,
Hellmann MarjaLeena,
Kjaer Susanne Krüger,
Fung Eric T.,
Høgdall Estrid
Publication year - 2008
Publication title -
proteomics – clinical applications
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.948
H-Index - 54
eISSN - 1862-8354
pISSN - 1862-8346
DOI - 10.1002/prca.200780010
Subject(s) - urine , reproducibility , chromatography , mass spectrometry , urine sample , sample preparation , sample (material) , chemistry , surface enhanced laser desorption/ionization , tandem mass spectrometry , protein mass spectrometry , biochemistry
Because of its non‐invasive sample collection method, human urine is an attractive biological material both for discovering biomarkers and for use in future screening trials for different diseases. Before urine can be used for these applications, standardized protocols for sample handling that optimize protein stability are required. In this explorative study, we examine the influence of different urine collection methods, storage temperatures, storage times, and repetitive freeze‐thaw procedures on the protein profiles obtained by surface‐enhanced laser desorption/ionization time‐of‐flight mass spectrometry (SELDI‐TOF‐MS). Prospectively collected urine samples from 11 women were collected as either morning or midday specimens. The effects of storage temperature, time to freezing, and freeze‐thaw cycles were assessed by calculating the number, intensity, and reproducibility of peaks visualized by SELDI‐TOF‐MS. On the CM10 array, 122 peaks were detected and 28 peaks were found to be significantly different between urine types, storage temperature and time to freezing. On the IMAC‐Cu array, 65 peaks were detected and 1 peak was found to be significantly different according to time to freezing. No significant differences were demonstrated for freeze‐thaw cycles. Optimal handling and storage conditions are necessary in clinical urine proteomic investigations. Collection of urine with a single and consistently performed protocol is needed to reduce analytical bias. Collecting only one urine type, which is stored for a limited period at 4°C until freezing at −80°C prior to analysis will provide the most stable profiles.