An Optimized Procedure for Proteomic Analysis of Extracellular Vesicles Using In-Stage Tip Digestion and DIA LC-MS/MS: Application to Liquid Biopsy in Cancer.
| Autor: | Soni RK; Herbert Irving Comprehensive Cancer Center, New York, NY, USA., Dimapanat L; Department of Pathology & Cell Biology, New York, NY, USA., Katari MS; Department of Biology, New York University, New York, NY, USA., Rai AJ; Herbert Irving Comprehensive Cancer Center, New York, NY, USA. ajr2170@cumc.columbia.edu.; Department of Pathology & Cell Biology, New York, NY, USA. ajr2170@cumc.columbia.edu.; Special Chemistry Laboratories, New York, NY, USA. ajr2170@cumc.columbia.edu. |
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| Jazyk: | angličtina |
| Zdroj: | Methods in molecular biology (Clifton, N.J.) [Methods Mol Biol] 2022; Vol. 2546, pp. 401-409. |
| DOI: | 10.1007/978-1-0716-2565-1_35 |
| Abstrakt: | Utilizing biofluids to identify cancer biomarkers has received considerable attention in the past decade. In this regard, serum and urine are convenient biofluids to noninvasively recapitulate information usually indicated by traditional tissue biopsies. In particular, we are interested in exploring the extracellular vesicle (ECV)-containing compartment of these fluids as a targeted source for cancer biomarker discovery. ECVs are membrane-enclosed particles, comprising of various fractions including exosomes, microvesicles, and apoptotic bodies. In both physiological and pathological states such as cancer, ECVs carry a rich load of molecular and protein cargoes, which aid in mediating intercellular communication between cells from various tissue types. Here we successfully enriched ECVs using a simple, low-cost, optimized method that we have developed; it is generalizable for the analysis of ECVs from multiple sample types. Such procedures are necessary as ECVs are nanoparticles that contain a treasure trove of large numbers of biomarkers each present at very low levels. Sample processing procedures can enrich for these vesicles and allow for the enhanced detection of proteins in downstream applications such as comprehensive proteomics methods using data-independent acquisition (DIA) and LC-MS/MS. (© 2022. The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.) |
| Databáze: | MEDLINE |
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