Pre-analytical operating procedures for serum Low Molecular Weight protein profiling
| Autor: | Francesco Di Girolamo, Paolo Somma, Fiorella Guadagni, Jhessica Alessandroni |
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| Rok vydání: | 2009 |
| Předmět: |
Adult
Male Proteomics Proteome Biophysics Peptide Biochemistry Hemolysis Biological specimen Young Adult Humans Low molecular weight protein Blood Coagulation Whole blood chemistry.chemical_classification Reproducibility Blood Specimen Collection Chromatography Molecular mass Chemistry Clinical Laboratory Techniques Reproducibility of Results Blood Proteins Blood Coagulation Factors Microspheres Molecular Weight Clotting time Spectrometry Mass Matrix-Assisted Laser Desorption-Ionization Metabolome Female |
| Zdroj: | Journal of proteomics. 73(3) |
| ISSN: | 1876-7737 |
| Popis: | Biological specimen collection and storage are an integral component of serum proteomics research. Although many efforts have been posed to address the effects of pre-analytical procedures, standardized protocols for collection and storage of samples for Low Molecular Weight (LMW) proteome profiling are still needed. Here we report a systematic analysis on the influence of pre-analytical factors [clotting times, temperature and time storage, addition of protease inhibitor (PI)] on serum LMW proteome profiling. Moreover, a comparison between manual versus automated peptide purification by functionalized magnetic bead-based MALDI-MS approach was performed. The results demonstrated best serum LMW proteins recovery and stability using a clotting time between 1 and 2 h, with serum stored up to 2 h either at room temperature or at 4 °C, independently of PI addition. PI addition to whole blood resulted in a lower number of LMW peaks detected. Finally, minimal effects on serum proteome profiles were observed after 1-month storage at − 80 °C, independently of PI addition on whole blood and/or serum. In conclusion, the use of standardized pre-analytical and storage procedures together with an automated peptide purification might minimize potential bias on serum LMW profiling results, thus allowing a better homogeneity and reproducibility in future proteomics studies. |
| Databáze: | OpenAIRE |
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