Novel therapeutic strategies are needed to control the SARS-CoV-2 (severe acute respiratory syndrome coronavirus 2) pandemic. Here, we present a protocol to anchor the SARS-CoV-2 spike (S-)protein in the cytoplasmic membranes of erythrocyte liposomes. A surfactant was used to stabilize the S-protein’s structure in the aqueous environment before insertion and to facilitate reconstitution of the S-proteins in the erythrocyte membranes. The insertion process was studied using coarse grained Molecular Dynamics (MD) simulations. Liposome formation and S-protein anchoring was studied by dynamic light scattering (DLS), ELV-protein co-sedimentation assays, fluorescent microcopy and cryo-TEM. The Erythro-VLPs (erythrocyte based virus like particles) have a well defined size of ∼200 nm and an average protein density on the outer membrane of up to ∼300 proteins/ μ m 2 . The correct insertion and functional conformation of the S-proteins was verified by dose-dependent binding to ACE-2 (angiotensin converting enzyme 2) in biolayer interferometry (BLI) assays. Seroconversion was observed in a pilot mouse trial after 14 days when administered intravenously, based on enzyme-linked immunosorbent assays (ELISA). This red blood cell based platform can open novel possibilities for therapeutics for the coronavirus disease (COVID-19) including variants, and other viruses in the future.
【초록키워드】 COVID-19, coronavirus disease, viruses, coronavirus, pandemic, protocol, Trial, Therapeutics, Dynamics, angiotensin converting enzyme 2, molecular dynamics, severe acute respiratory syndrome Coronavirus, virus, angiotensin converting enzyme, variants, ELISA, ACE-2, Protein, enzyme-linked immunosorbent assay, Seroconversion, enzyme-linked immunosorbent assays, surfactant, novel, respiratory, platform, therapeutic strategy, Blood, binding, SARS-CoV-2 spike, red blood cell, S-protein, angiotensin, open, blood cell, acute respiratory syndrome, Particles, acute respiratory syndrome coronavirus 2, enzyme, average, insertion, fluorescent, membranes, intravenously, outer membrane, Administered, BLI, Cell, defined, was used, virus, assays, facilitate, functional, dose-dependent, cytoplasmic membrane, scattering, the SARS-CoV-2, 【제목키워드】 SARS-CoV-2 spike protein, Liposome, anchoring,