{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,9]],"date-time":"2026-10-09T06:49:53Z","timestamp":1791528593632,"version":"4.3.4"},"reference-count":30,"publisher":"Wiley","license":[{"start":{"date-parts":[[2026,10,8]],"date-time":"2026-10-08T00:00:00Z","timestamp":1791417600000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"},{"start":{"date-parts":[[2026,10,8]],"date-time":"2026-10-08T00:00:00Z","timestamp":1791417600000},"content-version":"tdm","delay-in-days":0,"URL":"http:\/\/doi.wiley.com\/10.1002\/tdm_license_1.1"}],"content-domain":{"domain":["analyticalsciencejournals.onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Proteomics"],"abstract":"<jats:title>ABSTRACT<\/jats:title>\n                  <jats:p>Bead\u2010based enrichment strategies for low\u2010abundance proteins in biofluids, followed by LC\u2010MS\/MS analysis, have dramatically increased the coverage of the serum and plasma proteome. These technologies effectively mitigate the dynamic range limitations imposed by high\u2010abundance proteins like albumin, thereby facilitating the detection of low\u2010abundance proteins. Here, we demonstrate that bead\u2010based enrichment strategies are also disrupting the field of secretomics by enabling easy access to thousands of proteins released into the cell\u2010conditioned media, even in the presence of physiologically relevant growth supplements, such as fetal bovine serum. Using lipopolysaccharide\u2010stimulated THP\u20101 human acute monocytic leukemia cells as a model of inflammatory activation, we demonstrate that two independent bead\u2010based enrichment platforms, ENRICH\u2010iST and Proteograph, enable comprehensive characterization of secreted proteins in the conditioned media. Application of these approaches provided novel insights into the regulation of cytokine and chemokine signaling pathways that remained inaccessible without enrichment, even when employing labor\u2010intensive and time\u2010consuming deep fractionation workflows. Taken together, these findings underscore the potential of bead\u2010based enrichment methods to overcome long\u2010standing limitations in secretome analysis, offering a scalable and efficient route to uncover complex signaling networks with direct relevance to both fundamental biology and translational research.<\/jats:p>","DOI":"10.1002\/pmic.70190","type":"journal-article","created":{"date-parts":[[2026,10,9]],"date-time":"2026-10-09T06:01:01Z","timestamp":1791525661000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Bead\u2010Based Enrichment Methods for Unbiased Secretome Profiling Enable Unprecedented Protein Coverage in an Immune Cell Model"],"prefix":"10.1002","author":[{"given":"Barbara","family":"Kracher","sequence":"first","affiliation":[{"name":"Evotec International GmbH  Neuried Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Giada","family":"Marino","sequence":"additional","affiliation":[{"name":"Evotec International GmbH  Neuried Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Thomas","family":"Wild","sequence":"additional","affiliation":[{"name":"Evotec International GmbH  Neuried Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yannick","family":"Cogne","sequence":"additional","affiliation":[{"name":"Evotec (France) SAS Campus Curie  Toulouse CEDEX France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Till","family":"Kindel","sequence":"additional","affiliation":[{"name":"Evotec International GmbH  Neuried Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Marion","family":"Tondeur","sequence":"additional","affiliation":[{"name":"Evotec (France) SAS Campus Curie  Toulouse CEDEX France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Marie","family":"Guillemot","sequence":"additional","affiliation":[{"name":"Evotec (France) SAS Campus Curie  Toulouse CEDEX France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Andreas","family":"Tebbe","sequence":"additional","affiliation":[{"name":"Evotec International GmbH  Neuried Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Fran\u00e7ois","family":"Autelitano","sequence":"additional","affiliation":[{"name":"Evotec (France) SAS Campus Curie  Toulouse CEDEX France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Carleen","family":"Kluger","sequence":"additional","affiliation":[{"name":"Evotec International GmbH  Neuried Germany"},{"name":"Illumina GmbH  Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Samira","family":"Vautrin","sequence":"additional","affiliation":[{"name":"Evotec International GmbH  Neuried Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2026,10,8]]},"reference":[{"key":"e_1_2_12_2_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.mcpro.2023.100636"},{"key":"e_1_2_12_3_1","doi-asserted-by":"publisher","DOI":"10.1038\/nbt.2356"},{"key":"e_1_2_12_4_1","doi-asserted-by":"publisher","DOI":"10.1007\/978\u20101\u20100716\u20102863\u20108_19"},{"key":"e_1_2_12_5_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.cmet.2023.04.011"},{"key":"e_1_2_12_6_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41467\u2010020\u201017033\u20107"},{"key":"e_1_2_12_7_1","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.2106053119"},{"key":"e_1_2_12_8_1","doi-asserted-by":"crossref","unstructured":"T.Huang J.Wang A.Stukalov et\u00a0al. 2023. \u201cProtein Coronas on Functionalized Nanoparticles Enable Quantitative and Precise Large\u2010Scale Deep Plasma Proteomics. \u201d Preprint bioRxiv August 29 https:\/\/doi.org\/10.1101\/2023.08.28.555225.","DOI":"10.1101\/2023.08.28.555225"},{"key":"e_1_2_12_9_1","doi-asserted-by":"crossref","unstructured":"B.Lacar S.Ferdosi A.Alavi et\u00a0al. 2024. \u201cIdentification of Novel Biomarkers for Alzheimer's Disease and Related Dementias Using Unbiased Plasma Proteomics. \u201d Preprint bioRxiv January 8 https:\/\/doi.org\/10.1101\/2024.01.05.574446.","DOI":"10.1101\/2024.01.05.574446"},{"key":"e_1_2_12_10_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41598\u2010024\u201064316\u2010w"},{"key":"e_1_2_12_11_1","doi-asserted-by":"publisher","DOI":"10.1002\/prca.70000"},{"key":"e_1_2_12_12_1","doi-asserted-by":"publisher","DOI":"10.1021\/acs.jproteome.5c00221"},{"key":"e_1_2_12_13_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.jprot.2025.105519"},{"key":"e_1_2_12_14_1","doi-asserted-by":"publisher","DOI":"10.1038\/s43587\u2010025\u201000877\u20103"},{"key":"e_1_2_12_15_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.intimp.2014.08.002"},{"key":"e_1_2_12_16_1","doi-asserted-by":"publisher","DOI":"10.1016\/s0898\u20106568(00)00149\u20102"},{"key":"e_1_2_12_17_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41598\u2010023\u201041514\u20106"},{"key":"e_1_2_12_18_1","doi-asserted-by":"publisher","DOI":"10.1039\/c0fo00113a"},{"key":"e_1_2_12_19_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.mcpro.2024.100830"},{"key":"e_1_2_12_20_1","doi-asserted-by":"publisher","DOI":"10.1177\/17534259221100170"},{"key":"e_1_2_12_21_1","doi-asserted-by":"publisher","DOI":"10.3892\/mmr.2018.8542"},{"key":"e_1_2_12_22_1","doi-asserted-by":"publisher","DOI":"10.1177\/1753425907088244"},{"key":"e_1_2_12_23_1","doi-asserted-by":"crossref","unstructured":"I.Strielkov G.Buinovskaja A.Uvarovskii andV.Galatenko 2025. \u201cCutting Through the Clutter: Minimizing Redundancy in GO Enrichment Analysis With evoGO \u201d Preprint bioRxiv February 27 https:\/\/doi.org\/10.1101\/2025.02.24.639258.","DOI":"10.1101\/2025.02.24.639258"},{"key":"e_1_2_12_24_1","doi-asserted-by":"publisher","DOI":"10.1038\/nprot.2016.136"},{"key":"e_1_2_12_25_1","doi-asserted-by":"publisher","DOI":"10.1021\/acs.jproteome.2c00145"},{"key":"e_1_2_12_26_1","doi-asserted-by":"publisher","DOI":"10.1093\/nar\/gkl775"},{"key":"e_1_2_12_27_1","doi-asserted-by":"publisher","DOI":"10.3934\/biophy.2016.4.563"},{"key":"e_1_2_12_28_1","doi-asserted-by":"publisher","DOI":"10.1038\/nprot.2017.054"},{"key":"e_1_2_12_29_1","doi-asserted-by":"publisher","DOI":"10.1002\/eji.201040428"},{"key":"e_1_2_12_30_1","doi-asserted-by":"publisher","DOI":"10.1007\/s10753\u2010017\u20100601\u2010x"},{"key":"e_1_2_12_31_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.cellimm.2007.09.003"}],"container-title":["PROTEOMICS"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/analyticalsciencejournals.onlinelibrary.wiley.com\/doi\/pdf\/10.1002\/pmic.70190","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/analyticalsciencejournals.onlinelibrary.wiley.com\/doi\/full-xml\/10.1002\/pmic.70190","content-type":"application\/xml","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/analyticalsciencejournals.onlinelibrary.wiley.com\/doi\/pdf\/10.1002\/pmic.70190","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,10,9]],"date-time":"2026-10-09T06:01:02Z","timestamp":1791525662000},"score":1,"resource":{"primary":{"URL":"https:\/\/analyticalsciencejournals.onlinelibrary.wiley.com\/doi\/10.1002\/pmic.70190"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,10,8]]},"references-count":30,"alternative-id":["10.1002\/pmic.70190"],"URL":"https:\/\/doi.org\/10.1002\/pmic.70190","archive":["Portico"],"relation":{},"ISSN":["1615-9853","1615-9861"],"issn-type":[{"value":"1615-9853","type":"print"},{"value":"1615-9861","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,10,8]]},"assertion":[{"value":"2026-01-13","order":0,"name":"received","label":"Received","group":{"name":"publication_history","label":"Publication History"}},{"value":"2026-09-25","order":2,"name":"accepted","label":"Accepted","group":{"name":"publication_history","label":"Publication History"}},{"value":"2026-10-08","order":3,"name":"published","label":"Published","group":{"name":"publication_history","label":"Publication History"}}],"article-number":"e70190"}}