Article (Scientific journals)
Method validation for preparing urine samples for downstream proteomic and metabolomic applications.
Ammerlaan, Wim; Trezzi, Jean-Pierre; Mathay, Conny et al.
2014In Biopreservation and Biobanking, 12 (5), p. 351-7
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Keywords :
Adult; Cell-Derived Microparticles/metabolism; Cell-Free System; Creatinine/urine; Cystatins/urine; Female; Humans; Male; Metabolome; Middle Aged; Proteome; Reproducibility of Results; Temperature; Urine Specimen Collection/methods
Abstract :
[en] BACKGROUND: Formal validation of methods for biospecimen processing in the context of accreditation in laboratories and biobanks is lacking. A protocol for processing of a biospecimen (urine) was validated for fitness-for-purpose in terms of key downstream endpoints. METHODS: Urine processing was optimized for centrifugation conditions on the basis of microparticle counts at room temperature (RT) and at 4 degrees C. The optimal protocol was validated for performance (microparticle counts), and for reproducibility and robustness for centrifugation temperature (4 degrees C vs. RT) and brake speed (soft, medium, hard). Acceptance criteria were based on microparticle counts, cystatin C and creatinine concentrations, and the metabolomic profile. RESULTS: The optimal protocol was a 20-min, 12,000 g centrifugation at 4 degrees C, and was validated for urine collection in terms of microparticle counts. All reproducibility acceptance criteria were met. The protocol was robust for centrifugation at 4 degrees C versus RT for all parameters. The protocol was considered robust overall in terms of brake speeds, although a hard brake gave significantly fewer microparticles than a soft brake. CONCLUSIONS: We validated a urine processing method suitable for downstream proteomic and metabolomic applications. Temperature and brake speed can influence analytic results, with 4 degrees C and high brake speed considered optimal. Laboratories and biobanks should ensure these conditions are systematically recorded in the scope of accreditation.
Disciplines :
Life sciences: Multidisciplinary, general & others
Author, co-author :
Ammerlaan, Wim
Trezzi, Jean-Pierre ;  University of Luxembourg > Luxembourg Centre for Systems Biomedicine (LCSB)
Mathay, Conny
Hiller, Karsten ;  University of Luxembourg > Luxembourg Centre for Systems Biomedicine (LCSB)
Betsou, Fay
External co-authors :
yes
Language :
English
Title :
Method validation for preparing urine samples for downstream proteomic and metabolomic applications.
Publication date :
2014
Journal title :
Biopreservation and Biobanking
ISSN :
1947-5543
Publisher :
Mary Ann Liebert, United States - New York
Volume :
12
Issue :
5
Pages :
351-7
Peer reviewed :
Peer Reviewed verified by ORBi
Available on ORBilu :
since 16 May 2016

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