An integrated LC-ESI-MS platform for quantitation of serum peptide ladders. Application for colon carcinoma study.
Mounting evidence indicates that MS analysis of the human blood peptidome allows to distinguish between cancer and non-cancer samples, giving promise for a new MS-based diagnostic tool. However, several aspects of already published work have been criticized and demand for more methodical approach has been formulated. Motivated by this we undertook a systematic study of the plasma and serum peptidome using an integrated ESI-LC-MS-based platform, equipped with new data analysis tools for relative and absolute peptide quantitation. We used a high resolution LC-ESI-MS to analyze well-separated MS signals corresponding to peptides, and measured the variability of >1000 peptide signal amplitudes across a set of plasma and serum samples from healthy individuals. By spiking serum samples with known amounts of isotopically labeled versions of a selected set of peptides we measured the variability of their absolute concentration in this sample set and demonstrated a strong influence of clotting time on the concentration of these peptides in serum. Finally, we used this new LC-ESI-MS analytical platform for the differential analysis of healthy versus colon cancer serum samples and found that it was possible to distinguish the two groups with 89.8% sensitivity and 94.6% specificity.
Institute of Biochemistry and Biophysics, Polish Academy of Sciences, Warsaw, Poland.
Magda
M
Bakun
Jakub
J
Karczmarski
Jaroslaw
J
Poznanski
Tymon
T
Rubel
Malgorzata
M
Rozga
Agata
A
Malinowska
Dorota
D
Sands
Ewa
E
Hennig
Janusz
J
Oledzki
Jerzy
J
Ostrowski
Michal
M
Dadlez