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While the strong inflammatory properties of PGN and its derived muropeptides are well\u2010documented in human innate immune responses, adaptive immunity, including antibody responses to PGN, remain inadequately characterized. Microarray technology represents a cost\u2010 and time\u2010efficient method for studying such interactions. Our laser\u2010based technology enables the high\u2010throughput synthesis of biomolecules on functionalized glass slides. Here, this on\u2010chip synthesis was developed for PGN fragments, to generate a variety of 216 stem peptides and attach six different glycan moieties that are major structural components of bacterial cell walls. Thereby, 864 PGN fragments from different Gram\u2010negative and Gram\u2010positive species were generated. The arrays were validated with four different monoclonal antibodies against PGN or poly\u2010<jats:italic>N<\/jats:italic>\u2010acetyl glucosamine and identified their epitopes. Finally, proof of concept for antibody profiling in patient samples was performed by comparing a panel of well\u2010characterized plasma samples of epidermolysis bullosa (EB) patients suffering from (chronic) wounds with Staphylococcus aureus infection. EB patients show an increased response to the muramyl dipeptide. Therefore, this novel high\u2010throughput PGN glycopeptide microarray technology promises to identify distinct antibody profiles against human microbiomes in diseases, notably in those involving the intestine.<\/jats:p>","DOI":"10.1002\/anie.202420874","type":"journal-article","created":{"date-parts":[[2025,2,13]],"date-time":"2025-02-13T13:24:27Z","timestamp":1739453067000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":4,"title":["Synthetic High\u2010Throughput Microarrays of Peptidoglycan Fragments as a Novel Sero\u2010Diagnostic Tool for Patient Antibody Profiling"],"prefix":"10.1002","volume":"64","author":[{"given":"Alexandra","family":"Tsouka","sequence":"first","affiliation":[{"name":"Department of Biomolecular Systems Max Planck Institute of Colloids and Interfaces  14476 Potsdam Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yanyan","family":"Fu","sequence":"additional","affiliation":[{"name":"Department of Medical Microbiology and Infection Prevention University of Groningen University Medical Center Groningen  Groningen The Netherlands"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Manuel G.","family":"Ricardo","sequence":"additional","affiliation":[{"name":"Department of Biomolecular Systems Max Planck Institute of Colloids and Interfaces  14476 Potsdam Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Peter H.","family":"Seeberger","sequence":"additional","affiliation":[{"name":"Department of Biomolecular Systems Max Planck Institute of Colloids and Interfaces  14476 Potsdam Germany"},{"name":"Institute of Chemistry and Biochemistry Freie Universit\u00e4t Berlin  14195 Berlin Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yue","family":"Wang","sequence":"additional","affiliation":[{"name":"A*STAR Infectious Diseases Labs Agency for Science and Technology Research (A*STAR)  Singapore Singapore 138648"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Gerald B.","family":"Pier","sequence":"additional","affiliation":[{"name":"Mass General Brigham Harvard Medical School  Boston MA 02115 USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Detlef","family":"Schuppan","sequence":"additional","affiliation":[{"name":"Institute of Translational Immunology and Celiac Center Medical Center Johannes-Gutenberg University  55099 Mainz Germany"},{"name":"Division of Gastroenterology Beth Israel Deaconess Medical Center Harvard Medical School  Boston MA 02115 USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Louis","family":"Boon","sequence":"additional","affiliation":[{"name":"JJP Biologics  00-728 Warsaw Poland"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jan Maarten","family":"van Dijl","sequence":"additional","affiliation":[{"name":"Department of Medical Microbiology and Infection Prevention University of Groningen University Medical Center Groningen  Groningen The Netherlands"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Maria C.","family":"Bolling","sequence":"additional","affiliation":[{"name":"Department of Dermatology UMCG Center of Expertise for Blistering Diseases University Medical Center Groningen  The Netherlands"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Girbe","family":"Buist","sequence":"additional","affiliation":[{"name":"Department of Medical Microbiology and Infection Prevention University of Groningen University Medical Center Groningen  Groningen The Netherlands"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8227-2522","authenticated-orcid":false,"given":"Felix F.","family":"Loeffler","sequence":"additional","affiliation":[{"name":"Department of Biomolecular Systems Max Planck Institute of Colloids and Interfaces  14476 Potsdam Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jon D.","family":"Laman","sequence":"additional","affiliation":[{"name":"Department of Pathology and Medical Biology University of Groningen University Medical Center Groningen  Groningen The Netherlands"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"311","published-online":{"date-parts":[[2025,2,28]]},"reference":[{"key":"e_1_2_6_1_1","doi-asserted-by":"publisher","DOI":"10.1128\/br.36.4.407-477.1972"},{"key":"e_1_2_6_2_1","unstructured":"M.\u2005F. 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