Document Detail

Simple urinary sample preparation for proteomic analysis.
MedLine Citation:
PMID:  17022654     Owner:  NLM     Status:  MEDLINE    
Since the completion of the human genome sequence, attention has now focused on establishing reference maps of body fluids such as plasma and urine for detecting diagnostic markers of disease. Although some progress has been made, challenges still remain in the development of an optimal sample preparation method for proteomic analysis of urine. We have developed a simple and efficient urine preparation method for two-dimensional (2-D) gel electrophoresis which involves precipitation of proteins with simultaneous desalting. Acetonitrile precipitation produced 2-D gel separations with the highest resolution and the greatest number of protein spots compared to precipitation by other organic solvents. The method was applied to observe changes in the urinary proteome over a 6 week period and to establish a reference map of a healthy subject. A total of 339 proteins from 159 genes was identified from healthy male urine by peptide mass fingerprinting. The profiles of the urinary proteome at three times in 1 day and on four different days were compared and were found to vary in number and spatial location of the proteins on the map. The method was also shown to be applicable to the higher concentrations of protein found in the urine of an ovarian cancer subject. We have developed a facile and robust method for preparing urine for 2-D gels that will encourage further use of urine.
Alamgir Khan; Nicolle H Packer
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Publication Detail:
Type:  Journal Article    
Journal Detail:
Title:  Journal of proteome research     Volume:  5     ISSN:  1535-3893     ISO Abbreviation:  J. Proteome Res.     Publication Date:  2006 Oct 
Date Detail:
Created Date:  2006-10-06     Completed Date:  2006-12-18     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  101128775     Medline TA:  J Proteome Res     Country:  United States    
Other Details:
Languages:  eng     Pagination:  2824-38     Citation Subset:  IM    
Australian Proteome Analysis Facility (APAF), Level 4, Building F7B, Research Park Drive, Macquarie University, Sydney NSW 2109, Australia.
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MeSH Terms
Hydrogen-Ion Concentration
Neoplasm Proteins / urine
Ovarian Neoplasms / metabolism
Proteins / analysis*,  classification
Proteome / analysis*
Proteomics / methods*,  standards*
Urine / chemistry*
Reg. No./Substance:
0/Neoplasm Proteins; 0/Proteins; 0/Proteome

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