{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,7,30]],"date-time":"2025-07-30T11:59:24Z","timestamp":1753876764057,"version":"3.41.2"},"reference-count":14,"publisher":"Oxford University Press (OUP)","license":[{"start":{"date-parts":[[2021,6,14]],"date-time":"2021-06-14T00:00:00Z","timestamp":1623628800000},"content-version":"am","delay-in-days":0,"URL":"https:\/\/academic.oup.com\/journals\/pages\/open_access\/funder_policies\/chorus\/standard_publication_model"}],"funder":[{"DOI":"10.13039\/501100010785","name":"Canada First Research Excellence Fund","doi-asserted-by":"publisher","id":[{"id":"10.13039\/501100010785","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"abstract":"<jats:title>Abstract<\/jats:title>\n               <jats:p>Protein glycosylation is the attachment of a carbohydrate moiety to a protein backbone affecting both structure and function of the protein. Abnormal glycosylation is associated with various diseases, and some of the changes in glycosylation are detectable even before symptom development. As such, glycans have emerged as compelling new biomarker candidates. A wide range of analytical methods exist for small-scale glycan analyses. However, there is a growing need for highly robust and reproducible high-throughput techniques that allow for large-scale glycoprofiling. Here, we describe the evaluation of robustness and repeatability of immunoglobulin G (IgG) N-glycan analysis using the GlycoWorks RapiFluor-MS N-Glycan Kit followed by hydrophilic interaction ultra-high-performance liquid chromatography (HILIC-UHPLC) from 335 technical replicates of human plasma randomly distributed across 67 96-well plates. The data was collected over a 5-month period using multiple UHPLC systems and chromatographic columns. Following relative IgG N-glycan quantification in acquired chromatograms, data analysis showed that the most abundant peaks that together made up for three-fourths of the detected IgG N-glycome all had coefficients of variation (CVs) lower than 2%. The highest CVs ranging from 16 to 29% accompanied low abundance glycan peaks with the individual relative peak area below 1% that together made up for &amp;lt;2% of the detected IgG N-glycome. These results show that the tested method is very robust and repeatable, making it suitable for the IgG N-glycan analysis of a large number of samples in a high-throughput manner over a longer period of time.<\/jats:p>","DOI":"10.1093\/glycob\/cwab050","type":"journal-article","created":{"date-parts":[[2021,5,31]],"date-time":"2021-05-31T11:20:11Z","timestamp":1622460011000},"source":"Crossref","is-referenced-by-count":6,"title":["Robustness and repeatability of GlycoWorks RapiFluor-MS IgG <i>N<\/i>-glycan profiling in a long-term high-throughput glycomic study"],"prefix":"10.1093","author":[{"given":"Helena","family":"Deri\u0161","sequence":"first","affiliation":[{"name":"Genos Glycoscience Research Laboratory, Zagreb 10000, Croatia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Ana","family":"Cindri\u0107","sequence":"additional","affiliation":[{"name":"Genos Glycoscience Research Laboratory, Zagreb 10000, Croatia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Matthew","family":"Lauber","sequence":"additional","affiliation":[{"name":"Waters Corporation, 34 Maple Street, Milford, MA 01757-3696, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Tea","family":"Petrovi\u0107","sequence":"additional","affiliation":[{"name":"Genos Glycoscience Research Laboratory, Zagreb 10000, Croatia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Alicia","family":"Bielik","sequence":"additional","affiliation":[{"name":"New England Biolabs, 240 County Road, Ipswich, MA 01938-2723, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Christopher H","family":"Taron","sequence":"additional","affiliation":[{"name":"New England Biolabs, 240 County Road, Ipswich, MA 01938-2723, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Marleen","family":"van Wingerden","sequence":"additional","affiliation":[{"name":"Waters Corporation, 34 Maple Street, Milford, MA 01757-3696, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Gordan","family":"Lauc","sequence":"additional","affiliation":[{"name":"Genos Glycoscience Research Laboratory, Zagreb 10000, Croatia"},{"name":"Faculty of Pharmacy and Biochemistry, University of Zagreb, Zagreb 10000, Croatia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Irena","family":"Trbojevi\u0107-Akma\u010di\u0107","sequence":"additional","affiliation":[{"name":"Genos Glycoscience Research Laboratory, Zagreb 10000, Croatia"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"286","published-online":{"date-parts":[[2021,6,14]]},"reference":[{"key":"2021071416210360700_ref1","doi-asserted-by":"crossref","first-page":"2966","DOI":"10.1002\/1873-3468.13598","article-title":"N-glycome signatures in human plasma: Associations with physiology and major diseases","volume":"593","author":"Dotz","year":"2019","journal-title":"FEBS Lett"},{"key":"2021071416210360700_ref2","doi-asserted-by":"crossref","first-page":"65","DOI":"10.1016\/j.cellimm.2018.07.009","article-title":"Immunoglobulin G glycosylation in aging and diseases","volume":"333","author":"Gudelj","year":"2018","journal-title":"Cell Immunol"},{"key":"2021071416210360700_ref3","doi-asserted-by":"crossref","first-page":"1","DOI":"10.3389\/fchem.2018.00324","article-title":"Comparison of 2-aminobenzamide, procainamide and RapiFluor-MS as derivatizing agents for high-throughput HILIC-UPLC-FLR-MS N-glycan analysis","volume":"6","author":"Keser","year":"2018","journal-title":"Front Chem"},{"key":"2021071416210360700_ref4","doi-asserted-by":"crossref","first-page":"5401","DOI":"10.1021\/acs.analchem.5b00758","article-title":"Rapid preparation of released N-glycans for HILIC analysis using a labeling reagent that facilitates sensitive fluorescence and ESI-MS detection","volume":"87","author":"Lauber","year":"2015","journal-title":"Anal 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changes","volume":"18","author":"Reiding","year":"2019","journal-title":"Mol Cell Proteomics"},{"key":"2021071416210360700_ref8","doi-asserted-by":"crossref","DOI":"10.1080\/19420862.2020.1750794","article-title":"A rapid method for relative quantification of N-glycans from a therapeutic monoclonal antibody during trastuzumab biosimilar development","volume":"12","author":"Segu","year":"2020","journal-title":"MAbs"},{"key":"2021071416210360700_ref9","doi-asserted-by":"crossref","first-page":"37","DOI":"10.1016\/bs.mie.2016.09.027","article-title":"Comparative analysis and validation of different steps in glycomics studies","volume":"586","author":"Trbojevi\u0107-Akma\u010di\u0107","year":"2017","journal-title":"Methods Enzymol"},{"key":"2021071416210360700_ref11","doi-asserted-by":"crossref","first-page":"97","DOI":"10.1093\/glycob\/3.2.97","article-title":"Biological roles of oligosaccharides: All of the theories are correct","volume":"3","author":"Varki","year":"1993","journal-title":"Glycobiology"},{"key":"2021071416210360700_ref12","volume-title":"Essentials of Glycobiology","author":"Varki","year":"2009","edition":"2nd"},{"year":"2016","author":"Waters Corporation","key":"2021071416210360700_ref13"},{"year":"2017","author":"Waters Corporation","key":"2021071416210360700_ref14"},{"key":"2021071416210360700_ref15","doi-asserted-by":"crossref","first-page":"85","DOI":"10.1021\/acs.jproteome.9b00348","article-title":"Heritability of human plasma N-glycome","volume":"19","author":"Zaytseva","year":"2020","journal-title":"J Proteome 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