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. 2022 Sep 27;9:997305. doi: 10.3389/fmed.2022.997305

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Copyright © 2022 van der Pan, Kassem, Khatri, de Ru, Janssen, Tjokrodirijo, al Makindji, Stavrakaki, de Jager, Naber, de Laat, Louis, van den Bossche, Vogelezang, Balvers, Lamfers, van Veelen, Orfao, van Dongen, Teodosio and Díez.

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Experimental design. (A) Five different cell lysis buffers were combined with C18 and SP3 clean-up approaches to define 10 procedures, which were tested for their performance on quantity-limited samples (2,500 to 50,000 cells). (B) The combination of urea-based lysis buffer and SP3 was applied to study different monocytic subsets isolated from peripheral blood mononuclear cells (PBMCs) and macrophages from glioblastoma. Samples were labelled with TMT16-plex tags for quantitative analysis. cMo, classical monocytes; iMo, intermediate monocytes; LC-MS/MS, liquid chromatography-tandem mass spectrometry; ncMo, non-classical monocytes; SDS, sodium dodecyl sulfate; SP3, single-pot, solid-phase-enhanced sample preparation method; TEAB, triethylammonium bicarbonate; TFE, 2,2,2-trifluoroethanol; TMT, tandem mass tag [Created with BioRender.com].