biostudies-literatureKostas JCNCI NIH HHSDivision of Cancer Prevention, National Cancer InstituteNational Institute of General Medical SciencesNIGMS NIH HHSDana-Farber Cancer Institute/Northeastern University Joint Program in Cancer Drug Development Award1676-1688https://www.ebi.ac.uk/biostudies/studies/S-EPMC7954648biostudies-literatureUnknown20(3)In-depth LC-MS-based proteomic profiling of limited biological and clinical samples, such as rare cells or tissue sections from laser capture microdissection or microneedle biopsies, has been problematic due, in large, to the inefficiency of sample preparation and attendant sample losses. To address this issue, we developed on-microsolid-phase extraction tip (OmSET)-based sample preparation for limited biological samples. OmSET is simple, efficient, reproducible, and scalable and is a widely accessible method for processing ∼200 to 10,000 cells. The developed method benefits from minimal sample processing volumes (1-3 μL) and conducting all sample processing steps on-membrane within a single microreactor. We first assessed the feasibility of using micro-SPE tips for nanogram-level amounts Journal of proteome researchSimple and Efficient Microsolid-Phase Extraction Tip-Based Sample Preparation Workflow to Enable Sensitive Proteomic Profiling of Limited Samples (200 to 10,000 Cells).PMC7954648R01 CA218500R01GM120272R35 GM136421R01 GM120272R35GM136421R01CA218500Schejbal JIvanov ARKostas JCRay SGregus MfalseSimple and Efficient Microsolid-Phase Extraction Tip-Based Sample Preparation Workflow to Enable Sensitive Proteomic Profiling of Limited Samples (200 to 10,000 Cells).In-depth LC-MS-based proteomic profiling of limited biological and clinical samples, such as rare cells or tissue sections from laser capture microdissection or microneedle biopsies, has been problematic due, in large, to the inefficiency of sample preparation and attendant sample losses. To address this issue, we developed on-microsolid-phase extraction tip (OmSET)-based sample preparation for limited biological samples. OmSET is simple, efficient, reproducible, and scalable and is a widely accessible method for processing ∼200 to 10,000 cells. The developed method benefits from minimal sample processing volumes (1-3 μL) and conducting all sample processing steps on-membrane within a single microreactor. We first assessed the feasibility of using micro-SPE tips for nanogram-level amounts 2021-01-01T00:00:00Z2021 Mar2026-05-01T06:00:35.744Z2021-03-16T08:16:33ZS-EPMC79546483362586410.1021/acs.jproteome.0c00890