Multiplex Biomarker Screening Assay for Urinary Extracellular Vesicles Study: A Targeted LabelFree Proteomic Approach

被引：37
作者：
Chutipongtanate, Somchai [1 ,2 ]
Greis, Kenneth D. [1 ]
机构：
[1] Univ Cincinnati, Coll Med, Dept Canc Biol, UC Prote Lab, Cincinnati, OH 45220 USA
[2] Mahidol Univ, Ramathibodi Hosp, Fac Med, Pediat Translat Res Unit,Dept Pediat, Bangkok, Thailand
来源：
SCIENTIFIC REPORTS | 2018年 / 8卷
基金：
美国国家卫生研究院;
关键词：
DATA-INDEPENDENT ACQUISITION; EXOSOMES;
D O I：
10.1038/s41598-018-33280-7
中图分类号：
O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];
学科分类号：
07 ; 0710 ; 09 ;
摘要：
The recent advance in targeted label-free proteomics, SWATH-MS, can provide consistent protein detection and reproducible protein quantitation, which is a considerable advantage for biomarker study of urinary extracellular vesicles. We developed a SWATH-MS workflow with a curated spectral library of 1,145 targets. Application of the workflow across nine replicates of three sample types (exosomelike vesicles (ELVs), microvesicles (MVs) and urine proteins (UPs)) resulted in the quantitation of 888 proteins at FDR < 1%. The median-coefficient of variation of the 888 proteins in the ELV sample was 7.7%, indicating excellent reproducibility. Data analysis showed common exosome markers, (i.e. CD9, CD63, ALIX, TSG101 and HSP70) were enriched in urinary ELVs as compared to MVs and UPs. The use of a multiplex biomarker screening assay focused on ELVs was investigated, and perspectives in future applications are discussed.
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页数：8
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