tetano
Editor, Senior Moderator
PLoS One
. 2022 Mar 11;17(3):e0263671.
doi: 10.1371/journal.pone.0263671. eCollection 2022.
Erythro-VLPs: Anchoring SARS-CoV-2 spike proteins in erythrocyte liposomes
Sebastian Himbert[SUP] 1 2 [/SUP], Isabella Passos Gastaldo[SUP] 1 2 [/SUP], Rashik Ahmed[SUP] 3 4 [/SUP], Karla Martinez Pomier[SUP] 3 4 [/SUP], Braeden Cowbrough[SUP] 5 6 [/SUP], Dushyant Jahagirdar[SUP] 7 [/SUP], Samantha Ros[SUP] 3 [/SUP], Janos Juhasz[SUP] 1 8 [/SUP], Harald D H Stöver[SUP] 3 [/SUP], Joaquin Ortega[SUP] 7 [/SUP], Giuseppe Melacini[SUP] 3 4 [/SUP], Dawn M E Bowdish[SUP] 5 6 9 [/SUP], Maikel C Rheinstädter[SUP] 1 2 [/SUP]
Affiliations
Abstract
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/μm2. 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.
. 2022 Mar 11;17(3):e0263671.
doi: 10.1371/journal.pone.0263671. eCollection 2022.
Erythro-VLPs: Anchoring SARS-CoV-2 spike proteins in erythrocyte liposomes
Sebastian Himbert[SUP] 1 2 [/SUP], Isabella Passos Gastaldo[SUP] 1 2 [/SUP], Rashik Ahmed[SUP] 3 4 [/SUP], Karla Martinez Pomier[SUP] 3 4 [/SUP], Braeden Cowbrough[SUP] 5 6 [/SUP], Dushyant Jahagirdar[SUP] 7 [/SUP], Samantha Ros[SUP] 3 [/SUP], Janos Juhasz[SUP] 1 8 [/SUP], Harald D H Stöver[SUP] 3 [/SUP], Joaquin Ortega[SUP] 7 [/SUP], Giuseppe Melacini[SUP] 3 4 [/SUP], Dawn M E Bowdish[SUP] 5 6 9 [/SUP], Maikel C Rheinstädter[SUP] 1 2 [/SUP]
Affiliations
- PMID: 35275926
- PMCID: PMC8916654
- DOI: 10.1371/journal.pone.0263671
Abstract
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/μm2. 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.