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Multiplex Biomarker Screening Assay for Urinary Extracellular Vesicles Study: A Targeted Label-Free Proteomic Approach
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Multiplex Biomarker Screening Assay for Urinary Extracellular Vesicles Study: A Targeted Label-Free Proteomic Approach
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Journal Article
Multiplex Biomarker Screening Assay for Urinary Extracellular Vesicles Study: A Targeted Label-Free Proteomic Approach
2018
Overview
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 (exosome-like 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.
Publisher
Nature Publishing Group UK,Nature Publishing Group
Subject
/ 692/4022
/ 82/1
/ 82/29
/ 82/58
/ Calcium-Binding Proteins - metabolism
/ Cell Cycle Proteins - metabolism
/ Cell-Derived Microparticles - genetics
/ Cell-Derived Microparticles - metabolism
/ DNA-Binding Proteins - metabolism
/ Endosomal Sorting Complexes Required for Transport - metabolism
/ Extracellular Vesicles - genetics
/ Extracellular Vesicles - metabolism
/ HSP72 Heat-Shock Proteins - metabolism
/ Humanities and Social Sciences
/ Humans
/ Proteins
/ Science
/ Transcription Factors - metabolism
/ Urine
