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Protocol
Collection of urine samples for peptidome (e.g. LC-MS or CE-MS) analysis
Keywords: urine, mosaiques, CE-MS analysis
In order to facilitate the generation of meaningful data and to clarify the requirements for the analysis, we have prepared this sheet, indicating the most important steps.
Instructions
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Sampling
Urine 10 ml (minimum 2 ml). If more urine is collected, do not discard, please store it or ship it to us, as these samples may be valuable for MS/MS sequencing. Best if second morning urine, but everything between 7 and 11 am collection usually works fine. In generally mid stream urine should be collected, with exception of samples for prostate carcinoma studies (see below). 24h urine can be used as well, as long as it is collected in the presence of NaN3 (10 ml of a 5% solution) as preservative and hence does not contain microbial contamination. In general, samples should not be turbid and should be frozen without any further manipulation. Additives (e.g. protease inhibitors, chelating agents, etc.) must not be added.
If turbid samples cannot be avoided: spin if hemorrhagic or cellular contents are obvious (10 min, >3000 rpm, 4°C). However, such samples may result in data of lower quality.
Never spin sample after it has been stored for > 30 min in the cold, and certainly not after freezing!
First morning urine is not well suited, as it shows very high variability (probably due to differences in urinary output during the night). Urine obtained in afternoon and evening also shows a higher degree of variability, and generally lower concentration of peptides/proteins, which may result in the sample being of too low quality to be properly evaluated.
If first void urine is collected for PCa, aim to obtain the first 10 ml only, otherwise the data may not be useful (missing prostate secrete)!
Freeze sample in the urine monovette at -20°C minimum within 3 hours, avoid freeze thaw cycles!
Samples can be stored frozen at -20°C for years, if frost-free freezers (that undergo freeze thaw cycles) are not used. Automatic freeze/thaw cycles in freezers will compromise sample quality, must definitely be avoided. -
For all samples
- Patient ID and DATE of sample collection (or any other unique original sample label) needs to be on the vials (use appropriate marker!).
- Please send the medical history sheet together with the samples.
All samples should be frozen as soon as possible and stored on -20°C (or lower) until frozen shipment (usually on dry ice) to Mosaiques.
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Shipment
Please inform us of the carrier chosen and of the air bill number as soon as the samples are on the way.
Please feel free to contact us any time for more specific information at:
Mosaiques diagnostics and therapeutics Harald Mischak Mellendorfer Str. 7 30625 Hannover phone: +49-511-554744-13 or -22 or -0 fax: +49-511-554744-31
References
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Pilot study of capillary electrophoresis coupled to mass spectrometry as a tool to define potential prostate cancer biomarkers in urine. ,
Author: Theodorescu D, Fliser D, Wittke S, Mischak H, Krebs R, Walden M, Ross M, Eltze E, Bettendorf O, Wulfing C, Semjonow A | Date: Jul 2005 -
Discovery and validation of new protein biomarkers for urothelial cancer: a prospective analysis. ,
Author: Theodorescu D, Wittke S, Ross MM, Walden M, Conaway M, Just I, Mischak H, Frierson HF | Date: Mar 2006 -
CE-MS analysis of the human urinary proteome for biomarker discovery and disease diagnostics ,
Author: Coon JJ, Zürbig P, Dakna M, Dominiczak AF, Decramer S, Fliser D, Frommberger M, Golovko I, Good DM, Herget-Rosenthal S, Jankowski J, Julian BA, Kellmann M, Kolch W, Massy Z, Novak J, Rossing K, Schanstra JP, Schiffer E, Theodorescu D, Vanholder R, Weissinger EM, Mischak H, Schmitt-Kopplin P | Date: Juli 2008 -
Urinary proteomics in diabetes and CKD ,
Author: Rossing K, Mischak H, Dakna M, Zürbig P, Novak J, Julian BA, Good DM, Coon JJ, Tarnow L, Rossing P | Date: Juli 2008