title: Comparison of three different enrichment strategies for serum low molecular weight protein identification using shotgun proteomics approach.
abstract: Serum low-molecular weight (LMW) proteins potentially contain useful biological information and their identification can be used to discover novel potential biomarkers. Given the high complexity of serum samples, in the last years several different prefractionation and enrichment strategies have been developed. In this study three different methods, i.e. hydrogel nanoparticles, Proteominer(®) peptide ligand affinity beads and Sartorius Vivaspin(®) centrifugal ultrafiltration device, were compared and evaluated in order to select the best strategy for the enrichment and prefractionation of LMW proteins. A shotgun proteomics approach was adopted, with in-solution proteolytic digestion of the whole protein mixture and determination of the resulting peptides by nanoHPLC coupled with a high-resolution Orbitrap LTQ-XL mass spectrometer. Data analysis, focusing on the LMW proteome (MW ≤ 40 kDa), has shown that the hydrogel nanoparticles performed better in enriching the LMW protein profiles, with 115 proteins identified against 93 and 95 for Proteominer(®) beads and Sartorius Vivaspin(®) device, respectively.
authors: Dipartimento di Chimica, Sapienza Università di Roma, Rome, Italy. annalaura.capriotti@uniroma1.it
authors: Anna Laura
authors: AL
authors: Capriotti
authors: Giuseppe
authors: G
authors: Caruso
authors: Chiara
authors: C
authors: Cavaliere
authors: Susy
authors: S
authors: Piovesana
authors: Roberto
authors: R
authors: Samperi
authors: Aldo
authors: A
authors: Laganà
mesh_terms: D000328:Adult; D015415:Biomarkers / Q000097:blood; D001798:Blood Proteins / Q000032:analysis / Q000737:chemistry* / Q000302:isolation & purification*; D002498:Centrifugation; D006801:Humans; D020136:Hydrogel, Polyethylene Glycol Dimethacrylate / Q000737:chemistry; D008024:Ligands; D008297:Male; D008970:Molecular Weight; D053758:Nanoparticles / Q000737:chemistry; D010455:Peptides / Q000737:chemistry; D040901:Proteomics / Q000379:methods*; D012016:Reference Values; D013499:Surface Properties; D014462:Ultrafiltration; D055815:Young Adult
chemical_list: D015415:Biomarkers; D001798:Blood Proteins; D008024:Ligands; D010455:Peptides; D020136:Hydrogel, Polyethylene Glycol Dimethacrylate