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.
Highlights
Isolation and identification of low molecular weight proteins and peptides from urine is a critical need in biomedical research since they might represent a new source of biomarkers predictive of early-stage diseases [1,2]
We demonstrated the ability of functionalized core-shell silica nanoparticles (FCSNP) both to capture low molecular weight proteins and peptides by the pseudo-affinity hydrophobic molecular bait Cibacron blue, which interacts with proteins with sufficiently long hydrophobic regions, and to exclude high molecular weight proteins by the porous polymeric bis-acrylamide shell, all in a single step using model proteins [2]
The obtained FCSNP were synthesized in four stages and characterized in our previous report [2]
Summary
Isolation and identification of low molecular weight proteins and peptides from urine is a critical need in biomedical research since they might represent a new source of biomarkers predictive of early-stage diseases [1,2]. Isolation and purification of biomarkers is complicated because of low concentrations that are under the detection limits of mass spectrometry and conventional immunoassays [10,11,12] These proteins or peptides could be degraded by proteases or masked by the presence of different interfering substances or high molecular weight proteins such as albumin [5,12,13]. Many classes of nanoparticles such as quantum dots [15], silver and gold nanoparticles [16], hydrogels [17] and silica nanoparticles [18] have been proposed to be applicable in diagnosis, monitoring and treatment of diseases Among these known materials, silica nanoparticles may represent a good means to capture and isolate urinary proteins or peptides of interest. Overweight and obesity are two conditions of critical concern to global public health and are associated with numerous chronic diseases, including cardiovascular disease, chronic kidney disease, type 2 diabetes mellitus, hypertension and hyperlipidemia [23,24]
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