Serum Peptidome Profiling Revealed Platelet Factor 4 as a Potential Discriminating Peptide Associated with Pancreatic Cancer

• Published in: Clinical cancer research Vol. 15; no. 11; pp. 3812 - 3819
• Main Authors: FIEDLER, Georg Martin, BENEDIKT LEICHTLE, Alexander, HAUSS, Johann, BUCHLER, Markus, THIERY, Joachim, KASE, Julia, BAUMANN, Sven, CEGLAREK, Uta, FELIX, Klaus, CONRAD, Tim, WITZIGMANN, Helmut, WEIMANN, Arved, SCHÜTTE, Christof
• Format: Journal Article
• Language: English
• Published: Philadelphia, PA American Association for Cancer Research 01.06.2009
• Subjects: Antineoplastic agents; Biological and medical sciences; biomarker; Biomarkers, Tumor - blood; Blood Proteins - analysis; CA-19-9 Antigen - blood; Carcinoembryonic Antigen - blood; Diagnosis, Differential; Enzyme-Linked Immunosorbent Assay; Gastroenterology. Liver. Pancreas. Abdomen; Humans; Liver. Biliary tract. Portal circulation. Exocrine pancreas; MALDI-TOF MS; Medical sciences; pancreatic cancer; Pancreatic Neoplasms - blood; Pancreatic Neoplasms - diagnosis; Peptidome profiling; Pharmacology. Drug treatments; platelet factor 4; Platelet Factor 4 - blood; Proteomics - methods; Reproducibility of Results; Sensitivity and Specificity; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization - methods; Tumors; Platelet factor 4; Biological fluid; Peptides; Pancreas cancer; Digestive diseases; Serum; Malignant tumor; Cancer; Pancreatic disease
• ISSN: 1078-0432; 1557-3265
• DOI: 10.1158/1078-0432.CCR-08-2701

Purpose: Mass spectrometry–based serum peptidome profiling is a promising tool to identify novel disease-associated biomarkers, but is limited by preanalytic factors and the intricacies of complex data processing. Therefore, we investigated whether standardized sample protocols and new bioinformatic tools combined with external data validation improve the validity of peptidome profiling for the discovery of pancreatic cancer–associated serum markers. Experimental Design: For the discovery study, two sets of sera from patients with pancreatic cancer ( n = 40) and healthy controls ( n = 40) were obtained from two different clinical centers. For external data validation, we collected an independent set of samples from patients ( n = 20) and healthy controls ( n = 20). Magnetic beads with different surface functionalities were used for peptidome fractionation followed by matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry (MS). Data evaluation was carried out by comparing two different bioinformatic strategies. Following proteome database search, the matching candidate peptide was verified by MALDI-TOF MS after specific antibody-based immunoaffinity chromatography and independently confirmed by an ELISA assay. Results: Two significant peaks ( m / z 3884; 5959) achieved a sensitivity of 86.3% and a specificity of 97.6% for the discrimination of patients and healthy controls in the external validation set. Adding peak m / z 3884 to conventional clinical tumor markers (CA 19-9 and CEA) improved sensitivity and specificity, as shown by receiver operator characteristics curve analysis (AUROC combined = 1.00). Mass spectrometry–based m / z 3884 peak identification and following immunologic quantitation revealed platelet factor 4 as the corresponding peptide. Conclusions: MALDI-TOF MS-based serum peptidome profiling allowed the discovery and validation of platelet factor 4 as a new discriminating marker in pancreatic cancer.