112513 20240319080954.0 doi 10.3390/ijms23031150 sideral 128230 ART-2022-128230 eng Sancho-Albero, María (orcid)0000-0001-8762-5457 High-Performance Thin-Layer Chromatography-Densitometry-Tandem ESI-MS to Evaluate Phospholipid Content in Exosomes of Cancer Cells 2022 Access copy available to the general public Unrestricted The question of whether exosome lipids can be considered as potential cancer biomarkers faces our current limited knowledge of their composition. This is due to the difficulty in isolating pure exosomes, the variability of the biological sources from which they are extracted, and the uncertainty of the methods for lipid characterization. Here, we present a procedure to isolate exosomes and obtain a deep, repeatable, and rapid phospholipid (PL) composition of their lipid extracts, from embryonic murine fibroblasts (NIH-3T3 cell line) and none (B16-F1) and high (B16-F10) metastatic murine skin melanoma cells. The analytical method is based on High Performance Thin-Layer Chromatography with Ultraviolet and fluorescence densitometry and coupled to Electrospray (ESI)-tandem Mass Spectrometry (MS). Under the conditions described in this work, separation and determination of PL classes, (sphingomyelins, SM; phosphatidylcholines, PC; phosphatidylserines, PS; and phosphatidylethanolamines, PE) were achieved, expressed as mu g PL/100 mu g exosome protein, obtained by bicinchoninic acid assay (BCA). A detailed structural characterization of molecular species of each PL class was performed by simultaneous positive and negative ESI-MS and MS/MS directly from the chromatographic plate, thanks to an elution-based interface. info:eu-repo/grantAgreement/ES/CSIC/2021-80E076 info:eu-repo/grantAgreement/ES/DGA/E25-20R info:eu-repo/grantAgreement/EUR/ERC/ERC-AdG-742684 info:eu-repo/grantAgreement/ES/ISCIII-AES/PI21-00036 info:eu-repo/grantAgreement/ES/ISCIII/PI19-01007 info:eu-repo/semantics/openAccess by http://creativecommons.org/licenses/by/3.0/es/ 5.6 2022 BIOCHEMISTRY & MOLECULAR BIOLOGY 66 / 285 = 0.232 2022 Q1 T1 CHEMISTRY, MULTIDISCIPLINARY 52 / 178 = 0.292 2022 Q2 T1 1.154 2022 Medicine (miscellaneous) 2022 Q1 Physical and Theoretical Chemistry 2022 Q1 Computer Science Applications 2022 Q1 Inorganic Chemistry 2022 Q1 Spectroscopy 2022 Q1 Organic Chemistry 2022 Q1 Molecular Biology 2022 Q2 Catalysis 2022 Q2 7.8 2022 info:eu-repo/semantics/article info:eu-repo/semantics/publishedVersion Jarne, Carmen Saviróon, María (orcid)0000-0001-7299-0187 Martín-Duque, Pilar Universidad de Zaragoza (orcid)0000-0003-4160-9720 Membrado, Luis Cebolla, Vicente L. Santamaría, Jesús Universidad de Zaragoza (orcid)0000-0002-8701-9745 1013 275 Universidad de Zaragoza Dpto. Cirugía Área Estomatología 5005 555 Universidad de Zaragoza Dpto. Ing.Quím.Tecnol.Med.Amb. Área Ingeniería Química 23, 3 (2022), 1150 [14 pp] Int. j. mol. sci. International Journal of Molecular Sciences 1661-6596 1545487 http://zaguan.unizar.es/record/112513/files/texto_completo.pdf Versión publicada 2742479 http://zaguan.unizar.es/record/112513/files/texto_completo.jpg?subformat=icon icon Versión publicada oai:zaguan.unizar.es:112513 articulos driver 2024-03-18-13:22:12 ARTICLE