Pre-analytical operating procedures for serum Low Molecular Weight protein profiling

Francesco Di Girolamo, Jhessica Alessandroni, Paolo Somma, Fiorella Guadagni

Research output: Contribution to journalArticle

Abstract

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.

Original languageEnglish
Pages (from-to)667-677
Number of pages11
JournalJournal of Proteomics
Volume73
Issue number3
DOIs
Publication statusPublished - Jan 3 2010

Keywords

  • Low Molecular Weight (LMW) protein
  • Pre-analytical procedures
  • Proteomics
  • Serum
  • Storage

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics

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