Abstract
In this review, we report the evolution on experimental conditions for the analysis of normal urine based on combinatorial peptide ligand library (CPLL) treatment and successive 2-DE and 2-DE/MS analysis. The main topics are (i) definition of the urine sample requirements, (ii) optimization of the urine/ligand ratio, (iii) essay conditions, (iv) en bloc elution. Overall, normal urine protein composition as studied by 2-DE includes over 2600 spots. Relevant data on inter and intraessay reproducibility obtained by the analysis of different normal urines repeated several times are also here presented. We found a 73% reproducibility upon analysis of the same sample and 68% correspondence of protein composition among different normal urine samples. Based on the above results, we are completing the characterization with LC-MS of 249 spots. The composition of normal urine proteins after CPLLs is finally shown with the indication of those spots which are currently under identification. This map will be completed in a near future; in the meantime this would represent the basic reference sample for newly developed studies on human diseases.
Original language | English |
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Pages (from-to) | 509-515 |
Number of pages | 7 |
Journal | Proteomics |
Volume | 12 |
Issue number | 4-5 |
DOIs | |
Publication status | Published - Feb 2012 |
Keywords
- 2-DE
- Proteome maps
- Reproducibility
- Technology
ASJC Scopus subject areas
- Molecular Biology
- Biochemistry