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Nanoproteomic Approach for Isolation and Identification of Potential Biomarkers in Human Urine from Adults with Normal Weight,Overweight and Obesity
Authors:Sergio G Hernandez-Leon  Jose Andre-i Sarabia Sainz  Gabriela Ramos-Clamont Montfort  Jos ngel Huerta-Ocampo  Martha Nydia Ballesteros  Ana M Guzman-Partida  María del Refugio Robles-Burgueo  Luz Vazquez-Moreno
Abstract:In this work, previously synthesized and characterized core-shell silica nanoparticles (FCSNP) functionalized with immobilized molecular bait, Cibacron blue, and a porous polymeric bis-acrylamide shell were incubated with pooled urine samples from adult women or men with normal weight, overweight or obesity for the isolation of potential biomarkers. A total of 30 individuals (15 woman and 15 men) were included. FCSNP allowed the capture of a variety of low molecular weight (LMW) proteins as evidenced by mass spectrometry (MS) and the exclusion of high molecular weight (HMW) proteins (>34 kDa) as demonstrated by SDS-PAGE and 2D SDS-PAGE. A total of 36 proteins were successfully identified by MS and homology database searching against the Homo sapiens subset of the Swiss-Prot database. Identified proteins were grouped into different clusters according to their abundance patterns. Four proteins were found only in women and five only in men, whereas 27 proteins were in urine from both genders with different abundance patterns. Based on these results, this new approach represents an alternative tool for isolation and identification of urinary biomarkers.
Keywords:FCSNP  Cibacron blue  LMW proteins  urinary biomarkers  HMW proteins  nanoproteomics
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