Tsouka Alexandra, Fu Yanyan, Ricardo Manuel G, Seeberger Peter H, Wang Yue, Pier Gerald B, Schuppan Detlef, Boon Louis, van Dijl Jan Maarten, Bolling Maria C, Buist Girbe, Loeffler Felix F, Laman Jon D
Department of Biomolecular Systems, Max Planck Institute of Colloids and Interfaces, 14476, Potsdam, Germany.
Department of Medical Microbiology and Infection Prevention, University of Groningen, University Medical Center Groningen, Groningen, The Netherlands.
Angew Chem Int Ed Engl. 2025 Apr 25;64(18):e202420874. doi: 10.1002/anie.202420874. Epub 2025 Feb 28.
Peptidoglycan (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.
肽聚糖（PGN）是一种复杂的生物聚合物，对所有细菌物种的细胞壁完整性和功能至关重要。虽然PGN及其衍生的胞壁肽的强烈炎症特性在人类固有免疫反应中已有充分记录，但包括对PGN的抗体反应在内的适应性免疫仍未得到充分表征。微阵列技术是研究此类相互作用的一种经济高效的方法。我们基于激光的技术能够在功能化载玻片上高通量合成生物分子。在此，这种芯片上的合成方法被用于PGN片段，以生成各种216种主干肽，并连接六种不同的聚糖部分，这些是细菌细胞壁的主要结构成分。由此，产生了来自不同革兰氏阴性和革兰氏阳性物种的864个PGN片段。用四种针对PGN或聚N - 乙酰葡糖胺的不同单克隆抗体对这些阵列进行了验证，并确定了它们的表位。最后，通过比较一组患有金黄色葡萄球菌感染（慢性）伤口的大疱性表皮松解症（EB）患者的特征明确的血浆样本，对患者样本中的抗体谱进行了概念验证。EB患者对胞壁酰二肽的反应增强。因此，这种新型的高通量PGN糖肽微阵列技术有望识别针对疾病中人类微生物群的独特抗体谱，特别是在那些涉及肠道的疾病中。
