Open AccessErythro-VLPs: Anchoring SARS-CoV-2 spike proteins in erythrocyte liposomes
Author(s) -
Sebastian Himbert,
Isabella Passos Gastaldo,
Rashik Ahmed,
Karla Martinez Pomier,
Braeden Cowbrough,
Dushyant Jahagirdar,
Samantha Ros,
János Juhász,
Harald D. H. Stöver,
Joaquı́n Ortega,
Giuseppe Melacini,
Dawn M. E. Bowdish,
Maikel C. Rheinstädter
Publication year - 2022
Publication title -
plos one
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.99
H-Index - 332
ISSN - 1932-6203
DOI - 10.1371/journal.pone.0263671
Subject(s) - liposome , dynamic light scattering , chemistry , coronavirus , membrane , biophysics , membrane protein , virology , biology , microbiology and biotechnology , biochemistry , covid-19 , medicine , nanoparticle , materials science , nanotechnology , disease , pathology , infectious disease (medical specialty)
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.