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Here, we present the Enzyme\u2010Linked Cycloaddition Assay (ELCA), a click chemistry\u2010based platform for ultra\u2010sensitive detection and semi\u2010quantitative analysis of secreted sialoglycoproteins. By metabolically incorporating an azide\u2010modified sialic acid into newly synthesized glycoproteins and capturing labeled material via strain\u2010promoted cycloaddition, ELCA quantifies aggregate sialylation using a microplate reader\u2010compatible, ELISA\u2010like workflow. We demonstrate that the secreted glycoproteome responds rapidly to pharmacological perturbation, with changes detectable within hours. Benchmarking against common glycosylation inhibitors and profiling cytokine\u2010driven macrophage polarization further establishes ELCA's sensitivity and temporal resolution. Compatible with serum\u2010containing conditions and requiring no specialized instrumentation, ELCA provides a broadly accessible tool for rapid, cost\u2010effective monitoring of secreted glycoprotein dynamics.<\/jats:p>","DOI":"10.1002\/advs.78315","type":"journal-article","created":{"date-parts":[[2026,10,15]],"date-time":"2026-10-15T04:16:57Z","timestamp":1792037817000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Enzyme\u2010Linked Cycloaddition Assay (ELCA) for Rapid, Ultra\u2010Sensitive Monitoring of Secreted Sialoglycoproteins"],"prefix":"10.1002","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-2733-7124","authenticated-orcid":false,"given":"Jon","family":"Lundstr\u00f8m","sequence":"first","affiliation":[{"name":"Department of Chemistry and Molecular Biology University of Gothenburg  Gothenburg Sweden"},{"name":"Wallenberg Centre for Molecular and Translational Medicine University of Gothenburg  Gothenburg Sweden"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jingmei","family":"Yang","sequence":"additional","affiliation":[{"name":"Department of Chemistry and Molecular Biology University of Gothenburg  Gothenburg Sweden"},{"name":"Wallenberg Centre for Molecular and Translational Medicine University of Gothenburg  Gothenburg Sweden"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3008-7851","authenticated-orcid":false,"given":"Daniel","family":"Bojar","sequence":"additional","affiliation":[{"name":"Department of Chemistry and Molecular Biology University of Gothenburg  Gothenburg Sweden"},{"name":"Wallenberg Centre for Molecular and Translational Medicine University of Gothenburg  Gothenburg Sweden"},{"name":"Science For Life Laboratory (SciLifeLab) University of Gothenburg  Gothenburg Sweden"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2026,10,14]]},"reference":[{"key":"e_1_2_9_2_1","doi-asserted-by":"publisher","DOI":"10.1093\/glycob\/cww086"},{"key":"e_1_2_9_3_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.cell.2006.08.019"},{"key":"e_1_2_9_4_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41579\u2010018\u20100007\u20102"},{"key":"e_1_2_9_5_1","doi-asserted-by":"publisher","DOI":"10.3390\/biology10111178"},{"key":"e_1_2_9_6_1","doi-asserted-by":"publisher","DOI":"10.1038\/nrmicro2746"},{"key":"e_1_2_9_7_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.isci.2025.112605"},{"key":"e_1_2_9_8_1","doi-asserted-by":"publisher","DOI":"10.1155\/2015\/490531"},{"key":"e_1_2_9_9_1","doi-asserted-by":"publisher","DOI":"10.7150\/ijbs.93806"},{"key":"e_1_2_9_10_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.chembiol.2013.12.010"},{"key":"e_1_2_9_11_1","doi-asserted-by":"publisher","DOI":"10.1002\/mas.21730"},{"key":"e_1_2_9_12_1","doi-asserted-by":"publisher","DOI":"10.1042\/BST20200879"},{"key":"e_1_2_9_13_1","doi-asserted-by":"publisher","DOI":"10.1021\/acschembio.1c00689"},{"key":"e_1_2_9_14_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41592\u2010023\u201001841\u2010y"},{"key":"e_1_2_9_15_1","doi-asserted-by":"publisher","DOI":"10.1039\/C6SC01814A"},{"key":"e_1_2_9_16_1","doi-asserted-by":"publisher","DOI":"10.1038\/nbt.2356"},{"key":"e_1_2_9_17_1","doi-asserted-by":"publisher","DOI":"10.1021\/acs.analchem.0c05126"},{"key":"e_1_2_9_18_1","doi-asserted-by":"publisher","DOI":"10.1016\/0014\u20104827(84)90359\u20108"},{"key":"e_1_2_9_19_1","doi-asserted-by":"publisher","DOI":"10.1016\/0003-2697(84)90131-3"},{"key":"e_1_2_9_20_1","doi-asserted-by":"publisher","DOI":"10.1182\/blood.2024027038"},{"key":"e_1_2_9_21_1","doi-asserted-by":"publisher","DOI":"10.1038\/nri3737"},{"key":"e_1_2_9_22_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.cbpa.2024.102454"},{"key":"e_1_2_9_23_1","volume-title":"Essentials of Glycobiology","author":"Colley K. 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