Tsouka, AlexandraAlexandraTsoukaFu, YanyanYanyanFuRicardo, Manuel G.Manuel G.RicardoSeeberger, PeterPeterSeebergerWang, YueYueWangPier, Gerald B.Gerald B.PierSchuppan, DetlefDetlefSchuppanBoon, LouisLouisBoonVan Dijl Jan MaartenBolling, Maria C.Maria C.BollingBuist, GirbeGirbeBuistLoeffler, Felix F.Felix F.LoefflerLaman, Jon D.Jon D.Laman2025-11-102025-11-102025-04-25Angewandte Chemie International Edition 64 (18): e202420874 (2025)https://hdl.handle.net/11420/58562Peptidoglycan (PGN) is a complex biopolymer crucial for cell wall integrity and function of all bacterial species. While the strong inflammatory properties of PGN and its derived muropeptides are well-documented in human innate immune responses, adaptive immunity, including antibody responses to PGN, remain inadequately characterized. Microarray technology represents a cost- and time-efficient method for studying such interactions. Our laser-based technology enables the high-throughput synthesis of biomolecules on functionalized glass slides. Here, this on-chip 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-negative and Gram-positive species were generated. The arrays were validated with four different monoclonal antibodies against PGN or poly-N-acetyl glucosamine and identified their epitopes. Finally, proof of concept for antibody profiling in patient samples was performed by comparing a panel of well-characterized 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-throughput PGN glycopeptide microarray technology promises to identify distinct antibody profiles against human microbiomes in diseases, notably in those involving the intestine.en1521-3773202518antibodiesautoimmunityepidermolysis bullosa (EB)laser-induced forward transfer (LIFT)solid phase synthesisTechnology::600: TechnologyJournal Article10.1002/anie.202420874Journal Article