Enriched sera protein profiling for detection of non-small cell lung cancer biomarkers

Abstract

BackgroundNon Small Cell Lung Cancer (NSCLC) is the major cause of cancer related-death. Many patients receive diagnosis at advanced stage leading to a poor prognosis. At present, no satisfactory screening tests are available in clinical practice and the discovery and validation of new biomarkers is mandatory. Surface Enhanced Laser Desorption/Ionization Time-of-Flight Mass Spectrometry (SELDI-ToF-MS) is a recent high-throughput technique used to detect new tumour markers. In this study we performed SELDI-ToF-MS analysis on serum samples treated with the ProteoMiner™ kit, a combinatorial library of hexapeptide ligands coupled to beads, to reduce the wide dynamic range of protein concentration in the sample. Serum from 44 NSCLC patients and 19 healthy controls were analyzed with IMAC30-Cu and H50 ProteinChip Arrays.ResultsComparing SELDI-ToF-MS protein profiles of NSCLC patients and healthy controls, 28 protein peaks were found significantly different (p < 0.05), and were used as predictors to build decision classification trees. This statistical analysis selected 10 protein peaks in the low-mass range (2-24 kDa) and 6 in the high-mass range (40-80 kDa). The classification models for the low-mass range had a sensitivity and specificity of 70.45% (31/44) and 68.42% (13/19) for IMAC30-Cu, and 72.73% (32/44) and 73.68% (14/19) for H50 ProteinChip Arrays.ConclusionsThese preliminary results suggest that SELDI-ToF-MS protein profiling of serum samples pretreated with ProteoMiner™ can improve the discovery of protein peaks differentially expressed between NSCLC patients and healthy subjects, useful to build classification algorithms with high sensitivity and specificity. However, identification of the significantly different protein peaks needs further study in order to provide a better understanding of the biological nature of these potential biomarkers and their role in the underlying disease process.

SELDI-TOF-MS in chronic obstructive pulmonary disease

“…Our and Bowler's studies have demonstrated the feasibility of using SELDI as a diagnostic test for COPD, and that the serum protein fingerprint of COPD patients is different from that of healthy people. This permits to reveal low abundance proteins in serum and plasma samples, as needed for biomarker discovery [26][27][28]. However, our research results differ from those of Bowler's study.…”

Serum-based protein biomarkers for detection of lung cancer

“…The high-throughput nature and simplicity of its experimental procedures make SELDI-TOF MS a promising technology for future secretome analysis and biomarker discovery [47,48].…”

Mark Twain: How to fathom the depth of your pet proteome

“…However, if plasma is pre-treated with ProteoMiner, species can be detected down to as low as 10 pg protein per mL, with an increment of sensitivity of up to three orders of magnitude. Clear advantages in accessing the low-abundance plasma proteome via CPLL treatment have also been reported by Beseme et al [31] and other groups [16,[32][33][34][35][36][37][38] who claimed the importance of early-stage biomarkers. Although after a few years of use CPLLs did not yet generate validated protein markers, a number of scientific reports place a realistic discovery/validation hope very high.…”