EDTA-functionalized magnetic nanoparticles: A suitable platform for the analysis of low abundance urinary proteins

• Tipo: Artigo em Revista Científica Internacional
• Data: 2017

Autores

Bastos, P (Autor) Outra; Trindade, F (Autor) Outra; Ferreira, R (Autor) Outra; Leite-Moreira AF (Autor) FMUP; Falcao Pires, I (Autor) FMUP; Manadas, B (Autor) Outra; Daniel da Silva, AL (Autor) Outra; Vitorino, R (Autor) FMUP

Revista

Título: Talanta

Vol. 170

Páginas: 81-88

ISSN: 0039-9140

Editora: Elsevier

Indexação

ISI Web of Knowledge - 3 Citações

Scopus - 4 Citações

Outras Informações

ID Authenticus: P-00M-NDY

DOI: 10.1016/j.talanta.2017.03.087

Abstract (EN): Urine is a highly attractive source of biological information and disease biomarkers, whose proteome characterization is ongoing. To that end, depletion/enrichment strategies for protein analysis can be of great convenience. We have thus developed a method based on the use of EDTA-functionalized magnetic nanoparticles (NPs@EDTA), to fractionate urine samples before liquid chromatography-mass spectrometry analysis and compared the identified proteins with those obtained from ultrafiltrated/unfractionated (UF) urine samples. NPs@EDTA allowed larger urine volumes to be processed, resulting in a greater number of protein identifications (similar to 2-fold) at a lower cost when compared to UF samples. Proteins of greater abundance (such as albumin and uromodulin) were, at least partially, depleted with NPs@EDTA while those of lower abundance were enriched. Bioinformatics analysis showed that approximately 27% of NPs@EDTA-enriched proteins were annotated as displaying enzymatic activity, most of these being hydrolytic enzymes (56%), particularly proteases/peptidases (48%). Also, post-translational modifications were prominently predicted across NPs@ EDTA-enriched proteins (90%), particularly glycosylation (52%), phosphorylation (47%) and acetylation (30%). NPs@EDTA allowed the identification of 109 proteins in urine for the first time, showing high potential as a platform for urine's fractionation prior to proteomic analysis.

Idioma: Inglês

Tipo (Avaliação Docente): Científica

Nº de páginas: 8