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Several Anisakis excretory-secretory (E\/S) products and somatic proteins are known to be involved in IgE-mediated allergic reactions. In comparison with vertebrates, nematodes have a distinct machinery to glycosylate their proteins, and unusual glycan structures have been reported previously, many of which play immunogenic and immunomodulatory roles in host-parasite interactions. While an early study indicated that O-glycans participate in the cross-reactivity of antibodies in allergy patients to A. simplex somatic antigens, the N-glycosylation pattern of Anisakis and the potential role of N-glycans in allergic reactions remained unknown. The aim of the present study was to characterise N-glycans and the associated glycoproteins from Anisakis E\/S products using mass spectrometry. We collected E\/S products from larvae culture and released N-glycans from trypsinised proteins using PNGase Ar. Native glycans were pyridylaminated prior to HPLC separation and MALDI-TOF-MS\/MS analysis. In addition, hydrofluoric acid and glycosidase digestions were performed to aid structural characterisation. MS data of 5\u00a0h and 24\u00a0h E\/S products indicated the presence of pauci-mannose and core fucosylated N-glycans as major species; tri-fucosylated and methylated glycans as well as complex-type and phosphorylcholine-substituted glycans were also detected. In addition, E\/S products were subject to proteomics analysis, which revealed a set of proteins with conserved domains associated with allergens. Our study provides the first insight into the N-glycosylation machinery of Anisakis and highlights the need for investigating whether and which N-glycans are indubitably involved in the modulation of allergic responses.<\/jats:p>","DOI":"10.1042\/bcj20253281","type":"journal-article","created":{"date-parts":[[2026,2,26]],"date-time":"2026-02-26T11:26:54Z","timestamp":1772105214000},"page":"391-408","update-policy":"https:\/\/doi.org\/10.1042\/crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["Excretory\u2013secretory products of the fish-borne parasite\n                    <i>Anisakis simplex<\/i>\n                    L3 larvae possess allergens and unusual glycan modifications"],"prefix":"10.1042","volume":"483","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-9322-0130","authenticated-orcid":false,"given":"Isabella","family":"Adduci","sequence":"first","affiliation":[{"name":"1Institute of Parasitology, Department of Biological Sciences and Pathobiology, University of Veterinary Medicine Vienna, 1210 Vienna, Austria"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xiaoxu","family":"Wang","sequence":"additional","affiliation":[{"name":"2School of Biotechnology Jiangsu University of Science and Technology, Zhen Jiang, 212100, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3594-7136","authenticated-orcid":false,"given":"Katharina","family":"Paschinger","sequence":"additional","affiliation":[{"name":"3Institute of Biochemistry, University of Natural Resources and Life Sciences, Vienna, 1190 Vienna, Austria"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8996-1518","authenticated-orcid":false,"given":"Iain\u00a0B.H.","family":"Wilson","sequence":"additional","affiliation":[{"name":"3Institute of Biochemistry, University of Natural Resources and Life Sciences, Vienna, 1190 Vienna, Austria"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5249-1065","authenticated-orcid":false,"given":"Guofeng","family":"Cheng","sequence":"additional","affiliation":[{"name":"4Shanghai Tenth People\u2019s Hospital, Institute for Infectious Diseases and Vaccine Development, Tongji University School of Medicine, Shanghai, China"},{"name":"5Clinical Center for Brain and Spinal Cord Research, Tongji University, Shanghai 200092, China"},{"name":"6Affiliated Shanghai Blue Cross Brain hospital, School of Medicine, Tongji University, Shanghai 200020, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4949-9140","authenticated-orcid":false,"given":"Shi","family":"Yan","sequence":"additional","affiliation":[{"name":"1Institute of Parasitology, Department of Biological Sciences and Pathobiology, University of Veterinary Medicine Vienna, 1210 Vienna, Austria"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"288","published-online":{"date-parts":[[2026,3,18]]},"reference":[{"key":"2026031805562527000_B1","doi-asserted-by":"publisher","first-page":"43699","DOI":"10.1038\/srep43699","article-title":"Assessing the risk of an emerging zoonosis of worldwide concern: anisakiasis","volume":"7","author":"Bao","year":"2017","journal-title":"Sci. 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