Introduction: Proteomic analysis of formalin-fixed paraffin-embedded (FFPE) tumor tissue specimens has gained interest in the last 5 years due to technological advances and improved sample collection, as well as biobanking for clinical trials. The real-world implementation of clinical proteomics to these specimens, however, is hampered by tedious sample preparation steps and long instrument acquisition times.
Areas Covered: To advance the translation of quantitative proteomics into the clinic, we are comparing the performance of the leading commercial nanoflow liquid chromatography (nLC) system (based on literature reviews), the Easy-nLC 1200 (Thermo Fisher Scientific, Waltham, MA, U.S.A.), to the Evosep One HPLC (Evosep Biosystems, Odense, Denmark). We measured FFPE-tissue digests from 21 biological replicates with a similar gradient on both of the LC systems while keeping the on-column amount (1 µg total protein) and the single-shot data-dependent acquisition-based MS/MS method constant.
Expert Opinion: Overall, the Evosep One facilitates robust and sensitive high-throughput sample acquisition, making it suitable for clinical MS. We found the Evosep One to be a useful platform for positioning mass spectrometry-based proteomics in the clinical setting. The clinical application of nLC/MS will inform clinical decision-making in oncology and other diseases.
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http://dx.doi.org/10.1080/14789450.2023.2219844 | DOI Listing |
Mol Cell Proteomics
December 2024
Department of Biochemistry and Molecular Biophysics, Washington University School of Medicine, St. Louis, Missouri 63110, United States. Electronic address:
Histone post-translational modifications (PTMs) regulate gene expression patterns through epigenetic mechanisms. The 5 histone proteins (H1, H2A, H2B, H3, and H4) are extensively modified, with over 75 distinct modification types spanning more than 200 sites. Despite strong advances in mass spectrometry-based approaches, identification and quantification of modified histone peptides remains challenging due to factors such as isobaric peptides, pseudo-isobaric PTMs, and low stoichiometry of certain marks.
View Article and Find Full Text PDFElectrophoresis
November 2024
Faculty of Chemistry, Aalen University, Aalen, Germany.
In recent years, nanoflow sheath liquid (nanoSL) interfaces have been used more commonly for capillary electrophoresis-mass spectrometry (CE-MS) coupling due to their high sensitivity combined with flexibility in CE separation conditions. So far, the exact amount of sheath liquid (SL) and, thus, the dilution effect remain unknown due to the self-supplying type of the nanoSL interfaces. To quantify the SL flow rates, we present here an approach for the determination of the flow rate based on isotopically labeled standards using the nanoCEasy interface.
View Article and Find Full Text PDFPharmaceuticals (Basel)
November 2024
Department of Pharmacology and Neuroscience, University of North Texas Health Science Center, Fort Worth, TX 76107, USA.
Although estrogenic compounds promise therapeutic potential in treating various conditions, concerns regarding their endocrine-disrupting effects have been raised. Current methodologies for screening estrogenicity in rodent models are limited to the female-specific uterotrophic bioassay. Studies have reported enlargement of the seminal vesicles in orchiectomized males treated with estrogens.
View Article and Find Full Text PDFMol Cell Proteomics
December 2024
Department of Chemistry and Biochemistry, Brigham Young University, Provo, Utah, USA. Electronic address:
Mass spectrometry-based proteome profiling of trace analytes including single cells benefits from liquid chromatography separations operated at low flow rates (e.g., <50 nl/min).
View Article and Find Full Text PDFAnal Chim Acta
December 2024
University of Amsterdam, van 't Hoff Institute for Molecular Sciences, Analytical Chemistry Group, Science Park 904, 1098 XH, Amsterdam, the Netherlands; Centre of Analytical Sciences Amsterdam, Science Park 904, 1098 XH, Amsterdam, the Netherlands. Electronic address:
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