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However, direct visualization of lipid nanodomains remains challenging due to their nanoscale dimensions and dynamic behavior. Here, we combine expansion microscopy (ExM) with a fluorescent perfringolysin O domain\u20104 probe to map cholesterol\u2010rich nanodomains in the membrane of whole intact cells with a spatial resolution approaching \u223c40\u00a0nm on a confocal setup. We demonstrate that cholesterol\u2010rich domains predominantly localize to actin\u2010supported membrane protrusions in COS\u20107 cells. We directly visualize how the binding of therapeutic mAbs rituximab (RTX) and daratumumab (DARA) redistributes CD20 and CD38 on lymphoma and multiple myeloma cells, respectively, into cholesterol\u2010rich nanodomains. Accumulation of receptor\/mAb complexes within lipid nanodomains creates Fc fragment\u2010dense regions that enhance signal transduction, apoptotic signaling, and complement\u2010dependent cytotoxicity. Furthermore, RTX binding induces pronounced B\u2010cell polarization and accumulation of CD20\/RTX complexes in cholesterol\u2010rich membrane nanodomains, whereas CD38\/DARA complexes exhibit a more homogeneous membrane distribution indicating that mAb binding\u2010induced receptor reorganization occurs in a cell\u2010type\u2010 and receptor\u2010dependent manner at the nanoscale. Our results show how ExM can be used advantageously to improve our understanding of the complex interplay of mAbs and lipid nanodomains.<\/jats:p>","DOI":"10.1002\/anie.1206161","type":"journal-article","created":{"date-parts":[[2026,8,25]],"date-time":"2026-08-25T04:12:08Z","timestamp":1787631128000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Expansion Microscopy Reveals the Spatial Association of Therapeutic Antibody\u2013Receptor Complexes With Cholesterol\u2010Rich Membrane Domains"],"prefix":"10.1002","volume":"65","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-0121-1665","authenticated-orcid":false,"given":"Gang","family":"Wen","sequence":"first","affiliation":[{"name":"State Key Laboratory of Radiation Medicine and Protection School of Radiation Medicine and Protection Collaborative Innovation Center of Radiological Medicine of Jiangsu Higher Education Institutions Soochow University  Suzhou P. R. 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