{"doi":"10.1002/elps.201200657","title":"Enhanced glycan nanoprofiling by weak anion exchange preparative chromatography, mild acid desialylation, and nanoliquid chromatography‐mass spectrometry with nanofluorescence detection","abstract":"<jats:p>The structural characterization and quantification of the glycome of cells and glycoproteins is necessary for the understanding of glycan functions in Biology, the development of diagnostics tests, and the monitoring of glycoprotein pharmaceuticals. Classical <jats:italic><jats:styled-content style=\"fixed-case\">N</jats:styled-content></jats:italic>‐glycan characterization methods involve enzymatic release followed by derivatization with a fluorochrome and separation by normal‐phase <jats:styled-content style=\"fixed-case\">HPLC</jats:styled-content>. We have recently developed glycan nanoprofiling, a method for the simultaneous quantification and characterization of the <jats:italic><jats:styled-content style=\"fixed-case\">N</jats:styled-content></jats:italic>‐glycans without the need of external standardization. Although glycan nanoprofiling allows the characterization of both neutral and sialylated glycans within the same chromatographic run, a significant drawback is the coelution of similar glycans when complex glycan mixtures are analyzed. To overcome this problem, we have developed enhanced glycan nanoprofiling. This new method introduces a weak anion‐exchange <jats:styled-content style=\"fixed-case\">HPLC</jats:styled-content> separation step to fractionate glycans according to their sialic acid content followed by a mild acid desialylation. Glycans are then resolved by nano‐<jats:styled-content style=\"fixed-case\">LC</jats:styled-content>‐coupled <jats:styled-content style=\"fixed-case\">ESI</jats:styled-content>‐<jats:styled-content style=\"fixed-case\">MS</jats:styled-content> with an intercalated nanofluorescence detector. Neutral glycans have a better analytical separation, better ionization profiles, and provide significantly higher <jats:styled-content style=\"fixed-case\">MS</jats:styled-content> signals allowing a detailed characterization of rare glycan species. Enhanced glycan nanoprofiling is a powerful approach that provides a fast and sensitive alternative to available <jats:italic><jats:styled-content style=\"fixed-case\">N</jats:styled-content></jats:italic>‐glycan profiling methods.</jats:p>","journal":"ELECTROPHORESIS","year":2013,"id":595723,"datarank":0.40620753016533157,"base_score":2.70805020110221,"endowment":2.70805020110221,"self_citation_contribution":0.40620753016533157,"citation_network_contribution":0.0,"self_endowment_contribution":0.40620753016533157,"citer_contribution":0.0,"corpus_percentile":null,"corpus_rank":null,"citation_count":14,"citer_count":0,"citers_with_citation_signal":0,"citers_with_endowment":0,"datacite_reuse_total":0,"is_dataset":false,"is_dataset_confidence":null,"is_data_producer":false,"deposit_databanks":null,"is_oa":false,"file_count":0,"downloads":0,"has_version_chain":false,"published_date":null,"fair_score":null,"fair_percentile":null,"algorithm_id":"datarank_citation_only_1hop_v6","ranking_scope":"data_only","authors":[{"id":1238305,"name":"Martino Ambrosini","orcid":"0000-0003-1615-2984","position":1,"is_corresponding":false},{"id":990306,"name":"Fabrizio Chiodo","orcid":"0000-0003-3619-9982","position":2,"is_corresponding":false},{"id":931049,"name":"Yvette van Kooyk","orcid":"0000-0001-5997-3665","position":3,"is_corresponding":false},{"id":514769,"name":"Juan J. García‐Vallejo","orcid":"0000-0001-6238-7069","position":4,"is_corresponding":false},{"id":931576,"name":"Hakan Kalay","orcid":null,"position":0,"is_corresponding":false}],"reference_count":0,"raw_metadata":{"has_enrichment":true,"resolved":true,"title":"Enhanced glycan nanoprofiling by weak anion exchange preparative chromatography, mild acid desialylation, and nanoliquid chromatography‐mass spectrometry with nanofluorescence detection","abstract":"<jats:p>The structural characterization and quantification of the glycome of cells and glycoproteins is necessary for the understanding of glycan functions in Biology, the development of diagnostics tests, and the monitoring of glycoprotein pharmaceuticals. Classical <jats:italic><jats:styled-content style=\"fixed-case\">N</jats:styled-content></jats:italic>‐glycan characterization methods involve enzymatic release followed by derivatization with a fluorochrome and separation by normal‐phase <jats:styled-content style=\"fixed-case\">HPLC</jats:styled-content>. We have recently developed glycan nanoprofiling, a method for the simultaneous quantification and characterization of the <jats:italic><jats:styled-content style=\"fixed-case\">N</jats:styled-content></jats:italic>‐glycans without the need of external standardization. Although glycan nanoprofiling allows the characterization of both neutral and sialylated glycans within the same chromatographic run, a significant drawback is the coelution of similar glycans when complex glycan mixtures are analyzed. To overcome this problem, we have developed enhanced glycan nanoprofiling. This new method introduces a weak anion‐exchange <jats:styled-content style=\"fixed-case\">HPLC</jats:styled-content> separation step to fractionate glycans according to their sialic acid content followed by a mild acid desialylation. Glycans are then resolved by nano‐<jats:styled-content style=\"fixed-case\">LC</jats:styled-content>‐coupled <jats:styled-content style=\"fixed-case\">ESI</jats:styled-content>‐<jats:styled-content style=\"fixed-case\">MS</jats:styled-content> with an intercalated nanofluorescence detector. Neutral glycans have a better analytical separation, better ionization profiles, and provide significantly higher <jats:styled-content style=\"fixed-case\">MS</jats:styled-content> signals allowing a detailed characterization of rare glycan species. Enhanced glycan nanoprofiling is a powerful approach that provides a fast and sensitive alternative to available <jats:italic><jats:styled-content style=\"fixed-case\">N</jats:styled-content></jats:italic>‐glycan profiling methods.</jats:p>","is_dataset_classified":null,"base_score":2.70805020110221,"endowment":2.70805020110221,"datacite_reuse_total":0,"file_count":0,"downloads":0,"views":0,"has_version_chain":false,"is_dataset":false,"is_oa":false,"pmid":"23893432","pmcid":null,"openalex_id":"https://openalex.org/W2008437729","authors":[],"funders":[],"total_grants":0,"fwci":0.1475,"citation_percentile":0.55521268,"influential_citations":0,"citation_trend":[{"year":2013,"count":1},{"year":2017,"count":3},{"year":2018,"count":3},{"year":2019,"count":1},{"year":2020,"count":1},{"year":2022,"count":2},{"year":2023,"count":2},{"year":2024,"count":1}],"oa_status":"closed","license":"http://onlinelibrary.wiley.com/termsAndConditions#vor","oa_locations":[{"url":"https://api.wiley.com/onlinelibrary/tdm/v1/articles/10.1002%2Felps.201200657","host_type":"publisher"},{"url":"https://analyticalsciencejournals.onlinelibrary.wiley.com/doi/pdf/10.1002/elps.201200657","host_type":"publisher"},{"url":"https://doi.org/10.1002/elps.201200657","host_type":"journal"},{"url":"https://pubmed.ncbi.nlm.nih.gov/23893432","host_type":"repository"},{"url":"https://pure.amsterdamumc.nl/en/publications/381d0da4-80da-474f-8135-474226e1ce21","host_type":"repository"},{"url":"https://research.vumc.nl/en/publications/8ae1ade8-d287-447a-b45a-4c6d2c19059f","host_type":"repository"}],"fields_of_study":["Glycosylation and Glycoproteins Research","Carbohydrate Chemistry and Synthesis","Monoclonal and Polyclonal Antibodies Research"],"mesh_terms":["Animals","Anions","Carbohydrate Conformation","Cattle","Chromatography, High Pressure Liquid","Chromatography, Ion Exchange","Polysaccharides","Spectrometry, Fluorescence","N-Acetylneuraminic Acid","Nanotechnology","Tandem Mass Spectrometry","Glycomics","Fetuins"],"keywords":["Glycan","Chemistry","Chromatography","Glycomics","Glycome","Mass spectrometry","Sialic acid","Glycoprotein","Glycosylation","High-performance liquid chromatography","Derivatization","Biochemistry","Desialylation","Glycan Derivatization","Glycan Profiling","N-glycan Analysis"],"sdg_mappings":[],"linked_datasets":[],"clinical_trials":[],"software_tools":[],"database_accessions":[],"source":"live","citation_network_status":"fetched"},"created_at":"2026-07-27T17:49:11.375736Z","pmid":null,"pmcid":null,"fwci":null,"citation_percentile":null,"influential_citations":0,"oa_status":null,"license":null,"views":0,"total_file_size_bytes":0,"version_count":0,"fair_f":null,"fair_a":null,"fair_i":null,"fair_r":null,"fair_zscore":null,"fair_rationale":null,"fair_model":null,"fair_agent_version":null,"fair_fulltext_source":null,"fair_has_llm":null,"fair_computed_at":null,"clinical_trials":[],"software_tools":[],"db_accessions":[],"linked_datasets":[],"topics":[]}