Centro de Investigación en Alimentación y Desarrollo A.C., Carretera Gustavo Enrique Astiazarán Rosas, No. 46, col. La Victoria, Hermosillo, Sonora 83304, Mexico.
Departamento de Investigación en Física, Universidad de Sonora, Blvd. Luis Encinas y Rosales S/N, Col. Centro, Hermosillo, Sonora 83190, Mexico.
Molecules. 2021 Mar 23;26(6):1803. doi: 10.3390/molecules26061803.
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 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.
在这项工作中，我们使用先前合成并表征的核壳结构二氧化硅纳米颗粒（FCSNP），其表面固定有分子诱饵 Cibacron blue 和多孔聚合物双丙烯酰胺壳，与来自体重正常、超重或肥胖的成年男女的混合尿液样本一起孵育，以分离潜在的生物标志物。共有 30 人（15 名女性和 15 名男性）参与了该研究。FCSNP 允许捕获各种低分子量（LMW）蛋白质，这一点通过质谱（MS）得到了证实，而高分子量（HMW）蛋白质（>34 kDa）则被 SDS-PAGE 和 2D SDS-PAGE 排除在外。通过 MS 鉴定出 36 种蛋白质，并与 Swiss-Prot 数据库的子集进行同源性数据库搜索。根据丰度模式将鉴定出的蛋白质分为不同的簇。有 4 种蛋白质仅在女性尿液中发现，5 种仅在男性尿液中发现，而 27 种蛋白质在两性尿液中均有发现，但丰度模式不同。基于这些结果，这种新方法代表了一种用于分离和鉴定尿液生物标志物的替代工具。
