Reference : Method validation for preparing urine samples for downstream proteomic and metabolomi...
Scientific journals : Article
Life sciences : Multidisciplinary, general & others
Method validation for preparing urine samples for downstream proteomic and metabolomic applications.
Ammerlaan, Wim [> >]
Trezzi, Jean-Pierre mailto [University of Luxembourg > Luxembourg Centre for Systems Biomedicine (LCSB) >]
Mathay, Conny [> >]
Hiller, Karsten mailto [University of Luxembourg > Luxembourg Centre for Systems Biomedicine (LCSB) >]
Betsou, Fay [> >]
Biopreservation and biobanking
Yes (verified by ORBilu)
United States
[en] 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
[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.

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