{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,28]],"date-time":"2026-09-28T13:00:23Z","timestamp":1790600423903,"version":"4.1.0"},"reference-count":27,"publisher":"American Chemical Society (ACS)","issue":"9","license":[{"start":{"date-parts":[[2026,8,21]],"date-time":"2026-08-21T00:00:00Z","timestamp":1787270400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by-nc-nd\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100012166","name":"National Key Research and Development Program of China","doi-asserted-by":"publisher","award":["2017YFA0505901"],"award-info":[{"award-number":["2017YFA0505901"]}],"id":[{"id":"10.13039\/501100012166","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100012166","name":"National Key Research and Development Program of China","doi-asserted-by":"publisher","award":["2021YEE0200500"],"award-info":[{"award-number":["2021YEE0200500"]}],"id":[{"id":"10.13039\/501100012166","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100002858","name":"China Postdoctoral Science Foundation","doi-asserted-by":"publisher","award":["2017M610281"],"award-info":[{"award-number":["2017M610281"]}],"id":[{"id":"10.13039\/501100002858","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100003399","name":"Science and Technology Commission of Shanghai Municipality","doi-asserted-by":"publisher","award":["15002360172"],"award-info":[{"award-number":["15002360172"]}],"id":[{"id":"10.13039\/501100003399","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100012226","name":"Fundamental Research Funds for the Central Universities","doi-asserted-by":"publisher","award":["22120250317"],"award-info":[{"award-number":["22120250317"]}],"id":[{"id":"10.13039\/501100012226","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China","doi-asserted-by":"publisher","award":["92359203"],"award-info":[{"award-number":["92359203"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"publisher"}]},{"name":"Outstanding Clinical Discipline Project of Shanghai Pudong","award":["PWYgy2018-10"],"award-info":[{"award-number":["PWYgy2018-10"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2026,9,28]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>Cancer-associated defects in mucin-type O-glycosylation expose truncated glycans on cell-surface glycoproteins and create tumor-associated glycopeptide epitopes. However, the structural principles by which antibodies recognize specific MUC1 glycopeptide sequences remain incompletely defined. Here, we report the molecular basis of MUC1 recognition by 16A, a high-affinity antibody isolated against hypoglycosylated MUC1. Co-crystal structures of 16A Fab bound to site-selectively glycosylated MUC1 peptides revealed that 16A recognizes an STAPPAHG-centered epitope within the MUC1 variable number tandem repeat region. Surface plasmon resonance analysis showed that GalNAc modification at the threonine site in this sequence increased Fab binding affinity by 30.6-fold relative to the nonglycosylated peptide. Structural and mutational analyses identified a hydrogen bond between the threonine-linked GalNAc and Trp34 in heavy-chain CDR1 as a key contributor to this affinity enhancement. Cell-based binding assays further showed that COSMC deficiency enhanced 16A recognition of MUC1-expressing cells, while molecular dynamics simulations suggested that Ser-linked Core 1 extension can create local steric incompatibility near CDRH1 Arg32. Together, these results define a peptide-register-driven recognition mode in which the MUC1 peptide backbone determines specificity, whereas a site-compatible GalNAc acts as an affinity-enhancing element at the edge of the paratope. This structural framework may guide the design of MUC1 glycopeptide-based vaccines and antibody therapeutics.<\/jats:p>","DOI":"10.1021\/jacsau.6c00598","type":"journal-article","created":{"date-parts":[[2026,8,22]],"date-time":"2026-08-22T13:02:18Z","timestamp":1787403738000},"page":"5025-5036","source":"Crossref","is-referenced-by-count":0,"title":["Structural Basis of a High-Affinity Antibody Binding\nto Glycoprotein with Glycocalyx Decay"],"prefix":"10.1021","volume":"6","author":[{"given":"Yun","family":"Bin Han","sequence":"first","affiliation":[{"name":"ShanghaiTech University , , ,","place":["Shanghai, China, 201210"]},{"name":"Nanjing University of Chinese Medicine , , ,","place":["Nanjing, China, 210023"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jun","family":"Niu","sequence":"additional","affiliation":[{"name":"ShanghaiTech University , , ,","place":["Shanghai, China, 201210"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Deng","family":"Pan","sequence":"additional","affiliation":[{"name":"Tongji University School of Medicine , ,","place":["Shanghai, China, 200092"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Chunchao","family":"Feng","sequence":"additional","affiliation":[{"name":"Tongji University School of Medicine , ,","place":["Shanghai, China, 200092"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ke","family":"Song","sequence":"additional","affiliation":[{"name":"ShanghaiTech University , , ,","place":["Shanghai, China, 201210"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Bing","family":"Meng","sequence":"additional","affiliation":[{"name":"ShanghaiTech University , , ,","place":["Shanghai, China, 201210"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Shengyan","family":"Zhu","sequence":"additional","affiliation":[{"name":"Tongji University School of Medicine , ,","place":["Shanghai, China, 200092"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sandra","family":"Behren","sequence":"additional","affiliation":[{"name":"Ume\u00e5 University , , ,","place":["Ume\u00e5, Sweden, 90187"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ulrika","family":"Westerlind","sequence":"additional","affiliation":[{"name":"Ume\u00e5 University , , ,","place":["Ume\u00e5, Sweden, 90187"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4215-3684","authenticated-orcid":true,"given":"Yan","family":"Zhang","sequence":"additional","affiliation":[{"name":"Shanghai Jiao Tong University , , 800 Dongchuan Road , ,","place":["Shanghai, China, 200240"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Haiguang","family":"Liu","sequence":"additional","affiliation":[{"name":"Beijing Zhongguancun Academy , , ,","place":["Haidian, Beijing, China, 100094"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lan","family":"Xu","sequence":"additional","affiliation":[{"name":"ShanghaiTech University , , ,","place":["Shanghai, China, 201210"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1347-3811","authenticated-orcid":true,"given":"Dapeng","family":"Zhou","sequence":"additional","affiliation":[{"name":"Tongji University School of Medicine , ,","place":["Shanghai, China, 200092"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"316","published-online":{"date-parts":[[2026,8,21]]},"reference":[{"key":"2026092808081932200_cit1","doi-asserted-by":"publisher","first-page":"618","DOI":"10.1002\/prca.201300024","article-title":"Tn and\nSialyl-Tn Antigens: Aberrant O-Glycomics as Human Disease Markers","volume":"7","author":"Ju","year":"2013","journal-title":"Proteomics:Clin. Appl."},{"key":"2026092808081932200_cit2","doi-asserted-by":"publisher","first-page":"304","DOI":"10.1126\/science.1129200","article-title":"A Mutant Chaperone Converts a Wild-Type\nProtein into a Tumor-Specific Antigen","volume":"314","author":"Schietinger","year":"2006","journal-title":"Science"},{"key":"2026092808081932200_cit3","doi-asserted-by":"publisher","DOI":"10.3390\/biom6040039","article-title":"Intra- and Extra-Cellular\nEvents Related to Altered\nGlycosylation of MUC1 Promote Chronic Inflammation, Tumor Progression,\nInvasion, and Metastasis","volume":"6","author":"Cascio","year":"2016","journal-title":"Biomolecules"},{"key":"2026092808081932200_cit4","doi-asserted-by":"publisher","first-page":"5323","DOI":"10.1158\/1078-0432.CCR-09-0737","article-title":"The Prioritization\nof Cancer Antigens: A National Cancer Institute Pilot Project for\nthe Acceleration of Translational Research","volume":"15","author":"Cheever","year":"2009","journal-title":"Clin. Cancer Res."},{"key":"2026092808081932200_cit5","doi-asserted-by":"publisher","first-page":"2472","DOI":"10.2174\/13816128112092472","article-title":"Mucin-Based\nTargeted\nPancreatic Cancer Therapy","volume":"18","author":"Torres","year":"2012","journal-title":"Curr. Pharm. Des."},{"key":"2026092808081932200_cit6","doi-asserted-by":"publisher","first-page":"736","DOI":"10.1093\/glycob\/cwr182","article-title":"Control of Mucin-Type\nO-Glycosylation: A Classification\nof the Polypeptide GalNAc-Transferase Gene Family","volume":"22","author":"Bennett","year":"2012","journal-title":"Glycobiology"},{"key":"2026092808081932200_cit7","doi-asserted-by":"publisher","first-page":"150","DOI":"10.1021\/jacsau.3c00587","article-title":"Structure-Guided Approach for the Development of MUC1-Glycopeptide-Based\nCancer Vaccines with Predictable Responses","volume":"4","author":"Bermejo","year":"2024","journal-title":"JACS Au"},{"key":"2026092808081932200_cit8","doi-asserted-by":"publisher","first-page":"261","DOI":"10.1073\/pnas.1115166109","article-title":"Immune Recognition of Tumor-Associated Mucin MUC1 Is\nAchieved by a Fully Synthetic Aberrantly Glycosylated MUC1 Tripartite\nVaccine","volume":"109","author":"Lakshminarayanan","year":"2012","journal-title":"Proc. Natl. Acad. Sci. U.S.A."},{"key":"2026092808081932200_cit9","doi-asserted-by":"publisher","first-page":"1531","DOI":"10.1016\/S0968-0896(98)00092-3","article-title":"Effects of Glycosylation\non Fragments of Tumour Associated\nHuman Epithelial Mucin MUC1","volume":"6","author":"Braun","year":"1998","journal-title":"Bioorg. Med. Chem."},{"key":"2026092808081932200_cit10","doi-asserted-by":"publisher","first-page":"4981","DOI":"10.1002\/chem.200600144","article-title":"Synthesis\nand Structural Model of an \u03b1(2,6)-Sialyl-T Glycosylated MUC1\nEicosapeptide under Physiological Conditions","volume":"12","author":"Dziadek","year":"2006","journal-title":"Chem. - Eur. J."},{"key":"2026092808081932200_cit11","doi-asserted-by":"publisher","first-page":"7154","DOI":"10.1039\/C6CS00858E","article-title":"Principles of Mucin Structure: Implications\nfor the Rational Design of Cancer Vaccines Derived from MUC1-Glycopeptides","volume":"46","author":"Marti\u0301nez-Sa\u0301ez","year":"2017","journal-title":"Chem. Soc. Rev."},{"key":"2026092808081932200_cit12","doi-asserted-by":"publisher","DOI":"10.3390\/molecules23061326","article-title":"Epitopes of MUC1 Tandem Repeats in\nCancer as Revealed by Antibody Crystallography: Toward Glycopeptide\nSignature-Guided Therapy","volume":"23","author":"Zhou","year":"2018","journal-title":"Molecules"},{"key":"2026092808081932200_cit13","doi-asserted-by":"publisher","first-page":"9946","DOI":"10.1021\/bi012176z","article-title":"Effect of Glycosylation on MUC1 Humoral\nImmune Recognition:\nNMR Studies of MUC1 Glycopeptide-Antibody Interactions","volume":"41","author":"Grinstead","year":"2002","journal-title":"Biochemistry"},{"key":"2026092808081932200_cit14","doi-asserted-by":"publisher","first-page":"4999","DOI":"10.1039\/D0SC00317D","article-title":"A Straightforward Approach to Antibodies Recognising\nCancer Specific Glycopeptidic Neoepitopes","volume":"11","author":"Wakui","year":"2020","journal-title":"Chem.\nSci."},{"key":"2026092808081932200_cit15","doi-asserted-by":"publisher","first-page":"12438","DOI":"10.1021\/jacs.5b06787","article-title":"The Quest for Anticancer Vaccines: Deciphering the\nFine-Epitope Specificity of Cancer-Related Monoclonal Antibodies by\nCombining Microarray Screening and Saturation Transfer Difference\nNMR","volume":"137","author":"Coelho","year":"2015","journal-title":"J. Am. Chem. Soc."},{"issue":"24","key":"2026092808081932200_cit16","doi-asserted-by":"publisher","first-page":"13027","DOI":"10.1021\/jacs.2c12843","article-title":"MUC1 Glycopeptide\nVaccine Modified with a GalNAc Glycocluster Targets the Macrophage\nGalactose C-type Lectin on Dendritic Cells to Elicit an Improved Humoral\nResponse","volume":"145","author":"Gabba","year":"2023","journal-title":"J. Am. Chem. Soc."},{"issue":"5","key":"2026092808081932200_cit17","doi-asserted-by":"publisher","first-page":"2270","DOI":"10.1021\/jacsau.5c00224","article-title":"Enhancing Cancer Vaccine Efficacy: Backbone Modification\nwith \u03b2-Amino Acids Alters the Stability and Immunogenicity of\nMUC1-Derived Glycopeptide Formulations","volume":"5","author":"Gibadullin","year":"2025","journal-title":"JACS\nAu"},{"key":"2026092808081932200_cit18","doi-asserted-by":"publisher","first-page":"9830","DOI":"10.1002\/anie.201502813","article-title":"Deciphering the Non-Equivalence\nof Serine and Threonine O-Glycosylation Points: Implications for Molecular\nRecognition of the Tn Antigen by an Anti-MUC1 Antibody","volume":"54","author":"Marti\u0301nez-Sa\u0301ez","year":"2015","journal-title":"Angew. Chem., Int. Ed."},{"key":"2026092808081932200_cit19","doi-asserted-by":"publisher","first-page":"677","DOI":"10.1093\/glycob\/cww131","article-title":"Glycosylation of MUC1 Influences\nthe Binding of a Therapeutic Antibody by Altering the Conformational\nEquilibrium of the Antigen","volume":"27","author":"Movahedin","year":"2017","journal-title":"Glycobiology"},{"key":"2026092808081932200_cit20","doi-asserted-by":"publisher","first-page":"15137","DOI":"10.1039\/D0CC06349E","article-title":"Structural Characterization of an Unprecedented Lectin-like\nAntitumoral Anti-MUC1 Antibody","volume":"56","author":"Maci\u0301as-Leo\u0301n","year":"2020","journal-title":"Chem. Commun."},{"key":"2026092808081932200_cit21","doi-asserted-by":"publisher","first-page":"1977","DOI":"10.3892\/ijo.2012.1645","article-title":"MUC1 Glycopeptide Epitopes\nPredicted by Computational Glycomics","volume":"41","author":"Song","year":"2012","journal-title":"Int. J.\nOncol."},{"key":"2026092808081932200_cit22","doi-asserted-by":"publisher","first-page":"9458","DOI":"10.1021\/ja072181b","article-title":"Serine versus Threonine Glycosylation: The Methyl Group\nCauses a Drastic Alteration on the Carbohydrate Orientation and on\nthe Surrounding Water Shell","volume":"129","author":"Corzana","year":"2007","journal-title":"J. Am. Chem. Soc."},{"issue":"1","key":"2026092808081932200_cit23","doi-asserted-by":"publisher","first-page":"225","DOI":"10.1021\/jacsau.4c00921","article-title":"Decoding the Molecular Basis of the\nSpecificity of an Anti-sTn Antibody","volume":"5","author":"Soares","year":"2025","journal-title":"JACS Au"},{"key":"2026092808081932200_cit24","doi-asserted-by":"publisher","first-page":"10056","DOI":"10.1073\/pnas.0915176107","article-title":"Antibody Recognition of a Unique Tumor-Specific Glycopeptide\nAntigen","volume":"107","author":"Brooks","year":"2010","journal-title":"Proc. Natl. Acad. Sci. U.S.A."},{"key":"2026092808081932200_cit25","doi-asserted-by":"publisher","first-page":"223","DOI":"10.1107\/S2059798325002554","article-title":"Structure of the\nFab Fragment of a Humanized 5E5 Antibody to a Cancer-Specific Tn-MUC1\nEpitope","volume":"81","author":"Li","year":"2025","journal-title":"Acta Crystallogr., Sect. D:Struct.\nBiol."},{"key":"2026092808081932200_cit26","doi-asserted-by":"publisher","DOI":"10.1038\/s41467-019-08415-7","article-title":"Selection\nof Immunoglobulin Elbow Region Mutations Impacts Interdomain Conformational\nFlexibility in HIV-1 Broadly Neutralizing Antibodies","volume":"10","author":"Henderson","year":"2019","journal-title":"Nat. Commun."},{"key":"2026092808081932200_cit27","doi-asserted-by":"publisher","DOI":"10.1126\/sciadv.abg9118","article-title":"Cosmc Deficiency\nCauses Spontaneous Autoimmunity by Breaking B Cell Tolerance","volume":"7","author":"Zeng","year":"2021","journal-title":"Sci. Adv."}],"container-title":["JACS Au"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/pubs.acs.org\/doi\/pdf\/10.1021\/jacsau.6c00598","content-type":"application\/pdf","content-version":"vor","intended-application":"unspecified"},{"URL":"https:\/\/pubs.acs.org\/jaaucr\/article-pdf\/6\/9\/5025\/66987900\/jacsau.6c00598.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"syndication"},{"URL":"https:\/\/pubs.acs.org\/jaaucr\/article-pdf\/6\/9\/5025\/66987900\/jacsau.6c00598.pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,9,28]],"date-time":"2026-09-28T12:30:26Z","timestamp":1790598626000},"score":1,"resource":{"primary":{"URL":"https:\/\/pubs.acs.org\/jaaucr\/article\/6\/9\/5025\/5285768\/Structural-Basis-of-a-High-Affinity-Antibody"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,8,21]]},"references-count":27,"journal-issue":{"issue":"9","published-online":{"date-parts":[[2026,8,21]]},"published-print":{"date-parts":[[2026,9,28]]}},"URL":"https:\/\/doi.org\/10.1021\/jacsau.6c00598","relation":{},"ISSN":["2691-3704"],"issn-type":[{"value":"2691-3704","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,8,21]]}}}