Immunoconjugated Magnetic Graphene for Exosome Capture in SARS-CoV-2 Pseudovirus-Infected Cells

Graphene-based nanomaterials exhibit exceptional physicochemical properties that facilitate a range of diverse biomedical applications, including liquid biopsy. In this study, graphene-based magnetic units, termed MAGU (MAGnetic Units), were specifically engineered for the selective isolation of exosomes. Total extracellular vesicles were first enriched using ultracentrifugation, followed by immunomagnetic capture of CD9⁺ exosomes. MAGU functionalized with anti-CD9 antibody (MAGU-anti-CD9) efficiently recovered a CD9-positive exosome subpopulation expressing canonical markers ALIX, CD147, TSG101, and Flotillin-1, thereby confirming selective isolation performance. To investigate viral associated signaling, 293T cells were transduced with SARS-CoV-2 spike pseudovirus. This pseudovirus was engineered to express the SARS-CoV-2 spike protein, enabling simulation of viral entry and assessment of potential alterations in the exosomal profile induced by viral binding. Exosomes released by pseudovirus-transduced 293T cells were analyzed and compared to those from non-transduced controls. The MAGU-anti-CD9 complex selectively isolated a defined subset of CD9-positive vesicles enriched in the multifunctional transmembrane glycoprotein CD147, which has been proposed as a cofactor in SARS-CoV-2 entry. Comprehensive molecular profiling of selectively captured exosome subpopulations is expected to further support the application of MAGU technology in virus-host interaction research and liquid-biopsy-based diagnostics.