Standardized sample preparation phases for a quantitative measurement of plasma peptidome profiling by MALDI-TOF

Marco D'Imperio, Anna Della Corte, Angelo Facchiano, Michela Di Michele, Gabriella Ferrandina, Maria B. Donati, Domenico Rotilio

Research output: Contribution to journalArticlepeer-review

Abstract

A growing body of literature defines MALDI-TOF MS as a technique for studying plasma and serum, thus enabling the detection of proteins, and the generation of reproducible protein profile mass spectra, potentially able to discriminate correctly different biological systems. In this work, the different steps of the pre-analytical phase that may affect the reproducibility of plasma proteome analysis have been carefully considered. The results showed that the method is highly accurate (9.1%) and precise (8.9%) and the calibration curve for the ACTH (18-39), in human plasma, gave a good correlation coefficient (r > 0.99 and r2 > 0.98). The limit of detection (LOD) and the limit of quantification (LOQ), relative intensity, were of 0.5 × 10-9 M and 1.0 × 10-9 M respectively. Thus, an assay has been developed for the detection of low-abundant and low molecular weight proteins, from human plasma, aiming at the identification of new potential biomarkers. The method was tested on plasma from patients with a first diagnosis of pelvic mass. Statistical analysis of plasma profile generated a sub-profile of 17 peptides with their relative abundance able to discriminate patients bearing malignant or benign tumors. The sensitivity and specificity were 85.7% and 80.0% respectively.

Original languageEnglish
Pages (from-to)1355-1367
Number of pages13
JournalJournal of Proteomics
Volume73
Issue number7
DOIs
Publication statusPublished - May 7 2010

Keywords

  • Accuracy
  • Biomarkers
  • LOD-LOQ
  • MALDI-TOF MS
  • Plasma profile
  • Precision

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics

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