Peptidome profiling of induced sputum by mesoporous silica beads and MALDI-TOF MS for non-invasive biomarker discovery of chronic inflammatory lung diseases

• Published in: Proteomics (Weinheim) Vol. 11; no. 16; pp. 3402 - 3414
• Main Authors: Terracciano, Rosa, Preianò, Mariaimmacolata, Palladino, Grazia P., Carpagnano, Giovanna E., Barbaro, Maria P. Foschino, Pelaia, Girolamo, Savino, Rocco, Maselli, Rosario
• Format: Journal Article
• Language: English
• Published: Weinheim WILEY-VCH Verlag 01.08.2011; WILEY‐VCH Verlag; Wiley-VCH; Wiley Subscription Services, Inc
• Subjects: Adult; Aged; alpha-Defensins - metabolism; Analytical, structural and metabolic biochemistry; Asthma; Asthma - metabolism; Biological and medical sciences; biomarkers; Biomarkers - analysis; Biomarkers - chemistry; Case-Control Studies; Chronic obstructive pulmonary disease; Female; Fundamental and applied biological sciences. Psychology; high-throughput screening; Humans; Induced sputum; Inflammation; Inflammatory diseases; Leukocytes (neutrophilic); Lung diseases; MALDI-TOF MS; Male; Microspheres; Middle Aged; Miscellaneous; Molecular weight; Peptide; Peptides; Peptides - analysis; Peptides - chemistry; Peptidomics; Proteins; Proteome - analysis; Proteome - chemistry; proteomics; Proteomics - methods; Pulmonary Disease, Chronic Obstructive - metabolism; Reproducibility of Results; Respiratory diseases; Respiratory tract diseases; Saliva - chemistry; Saliva - metabolism; Serine; Silica; Silicon Dioxide - chemistry; Solid Phase Extraction; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization - methods; Sputum; Sputum - chemistry; Sputum - metabolism; Technology; Lung disease; Peptides; Respiratory disease; Chronic disease; Lung diseases; Biological marker; Peptidomics; Inflammatory disease; MALDI-TOF MS; Technology; Sputum; Proteomics; Peptide; Induced sputum
• ISSN: 1615-9853; 1615-9861; 1615-9861
• DOI: 10.1002/pmic.201000828

Induced sputum is recognized as being of increasing importance for the diagnosis and monitoring of chronic inflammatory lung diseases. The main purpose of this study is to provide a valid approach to better fractionate and characterize the still under‐estimated low‐molecular weight proteome of induced sputum by using mesoporous silica beads (MSBs) SPE coupled to MALDI‐TOF MS. Sputum peptides were captured from both derivatized and non‐derivatized MSBs and then profiled by MALDI‐TOF MS. Depending on the chemical groups present on the mesoporous surface, complex peptide mixtures were extracted from induced sputum and converted into reproducible MALDI profiles. The number of peaks detected as a function of S/N was evaluated for each mesoporous surface. More than 400 peaks with an S/N>5 were obtained in comparison to 200 peaks detected without MSBs. Additionally, as a proof‐of‐principle, we investigated the ability of this platform to discriminate between the “sputome” of patients with asthma and chronic obstructive pulmonary disease, and between these groups and those of healthy control subjects. Six m/z peaks emerged as potential diagnostic peptidic patterns able to differentiate these inflammatory airway diseases in the sputome range. Human α‐defensins (human neutrophil peptide (HNP)1, HNP2, HNP3) and three C‐terminal amidated peptides, one of which is phosphorylated on serine, were identified by MALDI‐TOF/TOF MS. These findings may contribute to defining a high‐throughput screening MS‐based platform for monitoring key peptidic‐biomarkers for inflammatory and chronic respiratory diseases in induced sputum samples.