Standardized sample preparation phases for a quantitative measurement of plasma peptidome profiling by MALDI-TOF.
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 r(2)>0.98). The limit of detection (LOD) and the limit of quantification (LOQ), relative intensity, were of 0.5 x 10(-)(9)M and 1.0 x 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.
Laboratory of Analytical Techniques and Proteomics, Research Laboratories, Catholic University, Campobasso, Italy. marco.dimperio@moli-sani.org
Marco
M
D'Imperio
Anna
A
Della Corte
Angelo
A
Facchiano
Michela
M
Di Michele
Gabriella
G
Ferrandina
Maria B
MB
Donati
Domenico
D
Rotilio
D001774:Blood Chemical Analysis / Q000379:methods* / Q000592:standards; D005591:Chemical Fractionation / Q000379:methods; D006801:Humans; D010449:Peptide Mapping / Q000379:methods* / Q000592:standards; D010455:Peptides / Q000097:blood*; D044367:Phase Transition; D020543:Proteome / Q000032:analysis*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D013048:Specimen Handling / Q000379:methods* / Q000592:standards; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods* / Q000592:standards; D014481:United States
D010455:Peptides; D020543:Proteome