{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,8]],"date-time":"2026-10-08T03:54:33Z","timestamp":1791431673705,"version":"4.3.3"},"reference-count":38,"publisher":"Wiley","issue":"19","license":[{"start":{"date-parts":[[2026,7,29]],"date-time":"2026-07-29T00:00:00Z","timestamp":1785283200000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"},{"start":{"date-parts":[[2026,7,29]],"date-time":"2026-07-29T00:00:00Z","timestamp":1785283200000},"content-version":"tdm","delay-in-days":0,"URL":"http:\/\/doi.wiley.com\/10.1002\/tdm_license_1.1"}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Small Methods"],"published-print":{"date-parts":[[2026,10]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n                  <jats:p>Aberrant glycosylation, particularly enhanced sialylation of membrane proteins, acts as a critical regulator of receptor signaling, immune evasion, and therapeutic resistance in cancer. However, systematic and multiplexed interrogation of protein\u2010specific sialylation in living cells remains technically challenging. Here, we report SATP\u2010seq, a next\u2010generation sequencing\u2010based strategy that integrates a sialic acid\u2010reactive probe (SA) with a targeted protein probe (TP) to enable ensemble profiling of protein\u2010specific sialylation. By combining metabolic glycan labeling with nanobody\u2010 or aptamer\u2010mediated protein recognition, SATP\u2010seq achieves dual recognition of sialic acids and protein epitopes through DNA\u2010programmed proximity ligation. The resulting ligation products encode sialylation states into unique DNA barcodes, converting glycosylation information into sequencable signals for multiplexed and quantitative analysis within a single sequencing run. Application of SATP\u2010seq to gefitinib\u2010sensitive PC9 and gefitinib\u2010resistant PC9GR cells enables parallel profiling of seven membrane glycoproteins and reveals resistance\u2010associated remodeling of protein\u2010specific sialylation. Notably, differential sialylation of EGFR and CD47 emerges between sensitive and resistant cells, suggesting coordinated reprogramming of proliferative and immune\u2010regulatory pathways. Enzymatic desialylation, EGFR knockdown, and proteomic validation collectively confirm the specificity and biological relevance of these alterations. Together, these findings establish multiplexed protein\u2010specific sialylation profiling as a scalable approach for dissecting glycosylation\u2010driven mechanisms of cancer drug resistance.<\/jats:p>","DOI":"10.1002\/smtd.70912","type":"journal-article","created":{"date-parts":[[2026,7,30]],"date-time":"2026-07-30T06:51:13Z","timestamp":1785394273000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Profiling Multiplexed Protein\u2010Specific Sialylation in Cancer Drug Resistance Using Proximity Ligation Sequencing"],"prefix":"10.1002","volume":"10","author":[{"ORCID":"https:\/\/orcid.org\/0009-0006-0212-8552","authenticated-orcid":false,"given":"Ning","family":"Zhang","sequence":"first","affiliation":[{"name":"School of Chemistry and Materials University of Science and Technology of China  Hefei Anhui China"},{"name":"Zhejiang Key Laboratory of Functional Nucleic Acids for Basic and Clinical Application Zhejiang Cancer Hospital, Hangzhou Institute of Medicine (HIM), Chinese Academy of Sciences  Hangzhou Zhejiang China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Gaoyu","family":"Song","sequence":"additional","affiliation":[{"name":"State Key Laboratory of Oral &amp; Maxillofacial Reconstruction and Regeneration, National Clinical Research Center for Oral Diseases, Shaanxi International Joint Research Center for Oral Diseases, Department of Oral Anatomy and Physiology, School of Stomatology The Fourth Military Medical University  Xi'an Shaanxi China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zhiyuan","family":"Peng","sequence":"additional","affiliation":[{"name":"Zhejiang Key Laboratory of Functional Nucleic Acids for Basic and Clinical Application Zhejiang Cancer Hospital, Hangzhou Institute of Medicine (HIM), Chinese Academy of Sciences  Hangzhou Zhejiang 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