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Among the bioactive molecules, proteins play critical roles in neonatal development and immune protection, notably due to the presence of glycosylations which support protein folding, stability, recognition, and host\u2013microbe interactions. Thus, glyco-associated proteome characterization in colostrum by mass spectrometry is plebiscite to elucidate its biological functions and therapeutic potential. In this review, we provide a comprehensive overview of MS-based workflows employed for colostrum glycoproteins analysis, highlighting commonly used strategies, methodological specificities, and the gathered biological information. We also discuss alternative analytical approaches that have been successfully applied to colostrum glycoprotein studies. Finally, we present a critical perspective on emerging technologies and unexplored workflows, evaluating their potential applicability, limitations, and future relevance for the analysis of glycosylation-associated proteins in colostrum.<\/jats:p>","DOI":"10.1021\/acs.jafc.6c06007","type":"journal-article","created":{"date-parts":[[2026,10,4]],"date-time":"2026-10-04T15:01:58Z","timestamp":1791126118000},"page":"31271-31289","source":"Crossref","is-referenced-by-count":0,"title":["Identification\nof Glycosylation-Associated Proteins\nin Colostrum: A Review of Mass Spectrometry-Based Strategies"],"prefix":"10.1021","volume":"74","author":[{"ORCID":"https:\/\/orcid.org\/0009-0000-3669-9958","authenticated-orcid":true,"given":"Isalyne","family":"Drewek","sequence":"first","affiliation":[{"name":"University of Mons - UMONS , , ,","place":["Mons, Belgium, B-7000"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0004-4488-4298","authenticated-orcid":true,"given":"Blandine","family":"Chazarin","sequence":"additional","affiliation":[{"name":"University of Mons - UMONS , , ,","place":["Mons, Belgium, B-7000"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2071-4310","authenticated-orcid":true,"given":"Ruddy","family":"Wattiez","sequence":"additional","affiliation":[{"name":"University of Mons - UMONS , , ,","place":["Mons, Belgium, B-7000"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"316","published-online":{"date-parts":[[2026,10,4]]},"reference":[{"key":"2026101408090563000_cit1","volume-title":"Nutritional and Immunological\nImportance of Colostrum for the New Born","author":"Kuralkar","year":"2010"},{"issue":"3","key":"2026101408090563000_cit2","doi-asserted-by":"publisher","first-page":"175","DOI":"10.1515\/jcim-2014-0039","article-title":"Bovine Colostrum:\nAn Emerging Nutraceutical","volume":"12","author":"Bagwe","year":"2015","journal-title":"J. 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Nutr."},{"key":"2026101408090563000_cit4","doi-asserted-by":"publisher","first-page":"198","DOI":"10.1038\/nature01511","article-title":"Mass Spectrometry-Based Proteomics","volume":"422","author":"Aebersold","year":"2003","journal-title":"Nature"},{"key":"2026101408090563000_cit5","doi-asserted-by":"crossref","first-page":"199","DOI":"10.1007\/978-3-030-15950-4_11","article-title":"Identification\nof Posttranslational Modifications (PTMs) of Proteins by Mass Spectrometry","volume-title":"Advancements of Mass Spectrometry in Biomedical Research","author":"Aslebagh","year":"2019"},{"issue":"1","key":"2026101408090563000_cit6","doi-asserted-by":"publisher","first-page":"429","DOI":"10.1146\/annurev-food-030216-030207","article-title":"Milk Glycans\nand Their Interaction with the Infant-Gut Microbiota","volume":"9","author":"Kirmiz","year":"2018","journal-title":"Annu. Rev. Food Sci. Technol."},{"key":"2026101408090563000_cit7","volume-title":"Immunology of Human\nMilk and Host Immunity","author":"Goldman","year":"2020"},{"issue":"7","key":"2026101408090563000_cit8","doi-asserted-by":"publisher","first-page":"1178","DOI":"10.1017\/S0007114516000234","article-title":"Levels\nof Innate Immune Factors in Preterm and Term\nMothers\u2019 Breast Milk during the 1st Month Postpartum","volume":"115","author":"Trend","year":"2016","journal-title":"Br. J. Nutr."},{"issue":"6","key":"2026101408090563000_cit9","doi-asserted-by":"publisher","first-page":"1139","DOI":"10.21037\/tp-22-469","article-title":"Exploring Dynamic\nChange and Influencing\nFactors of TGF-\u03921 and MUC1 in Human Breast Milk, and Their Association\nwith Infantile Diseases","volume":"12","author":"Wu","year":"2023","journal-title":"Transl. Pediatr."},{"issue":"2","key":"2026101408090563000_cit10","doi-asserted-by":"publisher","first-page":"vvaf202","DOI":"10.1093\/ijfood\/vvaf202","article-title":"Proteomic Approach-Based Absolute\nQuantification of\nLactadherin in Breast Milk Using Full-Length Expressed Stable Isotope-Labelled\nProtein Internal Standard with in-Gel Digestion","volume":"60","author":"Sato","year":"2025","journal-title":"Int. J. Food Sci. Technol."},{"issue":"6","key":"2026101408090563000_cit11","doi-asserted-by":"publisher","first-page":"2852","DOI":"10.1002\/vms3.1274","article-title":"Evaluation of the Differences in Proteomics of High-quality\nBovine Colostrum and Low-quality Bovine Colostrum","volume":"9","author":"Kac\u0327ar","year":"2023","journal-title":"Vet. Med. Sci."},{"issue":"1","key":"2026101408090563000_cit12","doi-asserted-by":"publisher","first-page":"265","DOI":"10.3390\/nu13010265","article-title":"Bovine Colostrum: Its Constituents and Uses","volume":"13","author":"Playford","year":"2021","journal-title":"Nutrients"},{"issue":"1","key":"2026101408090563000_cit13","doi-asserted-by":"publisher","first-page":"26","DOI":"10.1186\/s43014-022-00104-1","article-title":"Bioactive Compounds,\nNutritional Profile and Health Benefits of Colostrum: A Review","volume":"4","author":"Poonia","year":"2022","journal-title":"Food Prod., Process. 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Food Chem."},{"issue":"3","key":"2026101408090563000_cit19","doi-asserted-by":"publisher","first-page":"593","DOI":"10.1016\/j.cvfa.2004.06.005","article-title":"Managing the Production,\nStorage, and Delivery of Colostrum","volume":"20","author":"McGuirk","year":"2004","journal-title":"Vet. Clin.\nNorth Am. Food Anim. Pract."},{"issue":"3","key":"2026101408090563000_cit20","doi-asserted-by":"publisher","first-page":"535","DOI":"10.1016\/j.cvfa.2019.07.005","article-title":"Colostrum Management\nfor Dairy Calves","volume":"35","author":"Godden","year":"2019","journal-title":"Vet. Clin. North Am. Food\nAnim. 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Sports Sci."},{"issue":"3","key":"2026101408090563000_cit33","doi-asserted-by":"publisher","first-page":"1181","DOI":"10.1007\/s00394-017-1401-7","article-title":"A Low-Dose, 6-Week\nBovine Colostrum Supplementation Maintains Performance and Attenuates\nInflammatory Indices Following a Loughborough Intermittent Shuttle\nTest in Soccer Players","volume":"57","author":"Kotsis","year":"2018","journal-title":"Eur. J. Nutr."},{"issue":"6","key":"2026101408090563000_cit34","doi-asserted-by":"publisher","first-page":"197","DOI":"10.3390\/cosmetics11060197","article-title":"The Potential\nApplications of Natural Colostrum in Skin Health","volume":"11","author":"Bolat","year":"2024","journal-title":"Cosmetics"},{"key":"2026101408090563000_cit35","doi-asserted-by":"publisher","first-page":"1033779","DOI":"10.3389\/fnut.2022.1033779","article-title":"Role of Milk\nGlycome in Prevention, Treatment, and Recovery of COVID-19","volume":"9","author":"Kaplan","year":"2022","journal-title":"Front. Nutr."},{"issue":"6","key":"2026101408090563000_cit36","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pcbi.1006648","article-title":"Protein Ensembles\nLink Genotype to\nPhenotype","volume":"15","author":"Nussinov","year":"2019","journal-title":"PLoS Comput. Biol."},{"key":"2026101408090563000_cit37","article-title":"Mass Spectrometry\nfor Post-Translational Modifications","volume-title":"Neuroproteomics","author":"Parker","year":"2010"},{"issue":"11","key":"2026101408090563000_cit38","doi-asserted-by":"publisher","first-page":"6040","DOI":"10.1021\/acs.jafc.3c08957","article-title":"Phosphoproteomics Revealed\nDifferentially Expressed\nSites and Function of the Bovine Milk Fat Globule Membrane in Colostrum\nand Mature Milk","volume":"72","author":"Bai","year":"2024","journal-title":"J. Agric. Food Chem."},{"issue":"23","key":"2026101408090563000_cit39","doi-asserted-by":"publisher","first-page":"14677","DOI":"10.1021\/acs.jafc.5c02473","article-title":"Comparative\nPhosphorylation Dynamics of Whey Proteins in Human Colostrum and Mature\nMilk: Implications for Infant Nutrition","volume":"73","author":"Chen","year":"2025","journal-title":"J.\nAgric. Food Chem."},{"issue":"5","key":"2026101408090563000_cit40","doi-asserted-by":"publisher","first-page":"E127","DOI":"10.1038\/ncb0502-e127","article-title":"The Origins\nof Protein Phosphorylation","volume":"4","author":"Cohen","year":"2002","journal-title":"Nat. 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Sci."},{"key":"2026101408090563000_cit43","article-title":"O-Glycans","volume-title":"Essentials of Glycobiology","author":"Varki","year":"1999"},{"issue":"12","key":"2026101408090563000_cit44","doi-asserted-by":"publisher","first-page":"3096","DOI":"10.1016\/j.bbagen.2017.08.025","article-title":"Unusual Glycosylation\nof Proteins:\nBeyond the Universal Sequon and Other Amino Acids","volume":"1861","author":"Dutta","year":"2017","journal-title":"Biochim. Biophys. Acta, Gen. 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Proteomics"},{"issue":"3","key":"2026101408090563000_cit53","doi-asserted-by":"publisher","first-page":"1347","DOI":"10.3168\/jds.2012-5701","article-title":"Skimmed, Sterilized,\nand Concentrated Bovine Late Colostrum Promotes Both Prevention and\nRecovery from Intestinal Tissue Damage in Mice","volume":"96","author":"Cairangzhuoma","year":"2013","journal-title":"J. Dairy Sci."},{"issue":"9","key":"2026101408090563000_cit54","doi-asserted-by":"publisher","first-page":"665","DOI":"10.1007\/s10719-015-9615-5","article-title":"Lectin-Based Analysis of Fucosylated\nGlycoproteins of Human Skim Milk during 47 Days of Lactation","volume":"32","author":"Lis-Kuberka","year":"2015","journal-title":"Glycoconj. J."},{"issue":"38","key":"2026101408090563000_cit55","doi-asserted-by":"publisher","first-page":"10702","DOI":"10.1021\/acs.jafc.9b02034","article-title":"Mass Spectrometry Analysis\nof Changes in Human Milk N\/O -Glycopatterns\nat Different Lactation Stages","volume":"67","author":"Lu","year":"2019","journal-title":"J. Agric. Food\nChem."},{"key":"2026101408090563000_cit56","doi-asserted-by":"publisher","first-page":"1161310","DOI":"10.3389\/fnut.2023.1161310","article-title":"Changes in\nGlycosylated Proteins in Colostrum and Mature Milk and Their Implication","volume":"10","author":"Lu","year":"2023","journal-title":"Front. Nutr."},{"issue":"1","key":"2026101408090563000_cit57","doi-asserted-by":"publisher","first-page":"64","DOI":"10.2174\/1389203715666140221124622","article-title":"The Colostrum Proteome,\nRuminant Nutrition and Immunity: A Review","volume":"15","author":"Hernandez-Castellano","year":"2014","journal-title":"Curr.\nProtein Pept. 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