biostudies-literatureWang JHShanghai Municipal Science and Technology Major ProjectNational Key R&D Program of ChinaNational Natural Science Foundation of ChinaExcellent Research Program of Nanjing University0850https://www.ebi.ac.uk/biostudies/studies/S-EPMC12410930biostudies-literatureUnknown8O-glycopeptides are highly expressed in various human cancers and play a key role in cancer progression and metastasis, making them promising biomarkers for early diagnostics. However, the inherent complexity and heterogeneity of glycans pose a major challenge for the simultaneous and precise analysis of multiple glycopeptides. Here, we developed a low-temperature nanopore technique capable of simultaneously discriminating 4 truncated O-glycopeptides with varied glycoforms. This method enables the direct identification and relative quantification of O-glycopeptides from a mixture, achieving a discrimination accuracy of 92.9%. This general strategy holds promise for the label-free analysis of glycopeptide biomarkers, with potential applications in cancer diagnostics.Research (Washington, D.C.)Direct Identification of O-Glycopeptides by Low-Temperature Assisted Nanopore Technique.PMC1241093022334006ZYJH00422077130223771342022YFA1304604Li THu ZLGao ZWang JHYing YLMa WLong YTfalseDirect Identification of O-Glycopeptides by Low-Temperature Assisted Nanopore Technique.O-glycopeptides are highly expressed in various human cancers and play a key role in cancer progression and metastasis, making them promising biomarkers for early diagnostics. However, the inherent complexity and heterogeneity of glycans pose a major challenge for the simultaneous and precise analysis of multiple glycopeptides. Here, we developed a low-temperature nanopore technique capable of simultaneously discriminating 4 truncated O-glycopeptides with varied glycoforms. This method enables the direct identification and relative quantification of O-glycopeptides from a mixture, achieving a discrimination accuracy of 92.9%. This general strategy holds promise for the label-free analysis of glycopeptide biomarkers, with potential applications in cancer diagnostics.2025-01-01T00:00:00Z20252026-06-01T07:57:09.368Z2026-04-08T10:46:01.106ZS-EPMC124109304091749810.34133/research.0850