Hybrid Erythrocyte Liposomes: Hybrid Erythrocyte Liposomes: Functionalized Red Blood Cell Membranes for Molecule Encapsulation (Adv. Biosys. 3/2020)

Himbert, Sebastian; Blacker, Matthew J.; Kihm, Alexander; Pauli, Quinn; Khondker, Adree; Yang, Kevin; Sinjari, Sheilan; Johnson, Mitchell; Juhász, János; Wagner, Christian; Stöver, Harald D. H.; Rheinstädter, Maikel C.

doi:10.1002/adbi.202070031

Cited by 4 publications

(8 citation statements)

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“…Remaining Triton-X 100 micelles were detected between an elution volume of 15 ml and 30 ml, showing that the Amberlite XAD-2 beads do not completely remove Triton-X 100 and subsequent separation with SEC is essential for the purification of the Erythro-VLPs. The size distribution of the purified erythrocyte liposomes with and without S-protein was determined by dynamic light scattering (DLS) and is shown in Fig 2B . While erythrocyte liposomes measured 102 nm (polydispersity: 0.19), in good agreement with previous results [35], an average diameter of 222 nm (polydispersity: 0.32) was determined for the Erythro-VLPs.…”

Section: Plos Onesupporting

confidence: 91%

“…10 mg/ml 1-Palmitoyl-2-Oleoyl-sn-Glycero-3-Phosphocholin (POPC) in chloroform was prepared containing 1 mol% TR-DHPE. POPC has been previously shown to homogenously fuse with RBC membranes [35] and facilitates the incorporation of stained lipids into the membrane. 50 μL of this…”

Section: Staining Of Erythro-vlpsmentioning

confidence: 99%

“…Erythrocyte liposomes where first prepared according to the protocol described above. 10 samples, 1 ml each, containing *0.5 mg/ml of membrane material were prepared with Triton-X 100 at concentrations of 0.1, 1, 3,5,10,15,20,25,30,35 mM. The size distribution of the liposomes and Triton-X 100 micelles were then determined using a Zetasizer Nano ZS from Malvern Panalytical.…”

Section: Determining the Triton-x 100-erythrocyte Liposome Phase Diagrammentioning

confidence: 99%

“…MD simulations were performed on a GPU accelerated computer workstation using GRO-MACS Version 5.1.4. The device is equipped with a 40 Core central processing unit (CPU, Intel(R) Xeon(R) CPU E5-2630 v4 @ 2.20GHz), 130 GB random-access memory (RAM) and three graphic processing units (GPU, 2 × NVIDIA 1080 TDI + 1 × GeForce GT 730) [35]. A total of 4 simulation systems were created.…”

Section: Molecular Dynamics Simulationsmentioning

confidence: 99%

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Erythro-VLPs: Anchoring SARS-CoV-2 spike proteins in erythrocyte liposomes

et al. 2022

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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.

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Section: Plos Onesupporting

confidence: 91%

Section: Staining Of Erythro-vlpsmentioning

confidence: 99%

Section: Determining the Triton-x 100-erythrocyte Liposome Phase Diagrammentioning

confidence: 99%

Section: Molecular Dynamics Simulationsmentioning

confidence: 99%

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Erythro-VLPs: Anchoring SARS-CoV-2 spike proteins in erythrocyte liposomes

et al. 2022

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“…In addition, fresh blood was collected from volunteers in 10 ml heparinized blood collection tubes. Hemoglobin depleted RBC liposomes were then prepared from all samples following a previously published protocol [29,30]. Briefly: The whole blood was washed twice and the RBCs were isolated by successive centrifugation and replacing the supernatant with phosphate saline buffer (PBS).…”

Section: Preparation Of Solid Supported Rbc Cytoplasmic Membranesmentioning

confidence: 99%

Blood bank storage of red blood cells increases RBC cytoplasmic membrane order and bending rigidity

et al. 2021

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Blood banks around the world store blood components for several weeks ensuring its availability for transfusion medicine. Red blood cells (RBCs) are known to undergo compositional changes during storage, which may impact the cells’ function and eventually the recipients’ health. We extracted the RBC’s cytoplasmic membrane (RBCcm) to study the effect of storage on the membranes’ molecular structure and bending rigidity by a combination of X-ray diffraction (XRD), X-ray diffuse scattering (XDS) and coarse grained Molecular Dynamics (MD) simulations. Blood was stored in commercial blood bags for 2 and 5 weeks, respectively and compared to freshly drawn blood. Using mass spectrometry, we measured an increase of fatty acids together with a slight shift towards shorter tail lengths. We observe an increased fraction (6%) of liquid ordered (lo) domains in the RBCcms with storage time, and an increased lipid packing in these domains, leading to an increased membrane thickness and membrane order. The size of both, lo and liquid disordered (ld) lipid domains was found to decrease with increased storage time by up to 25%. XDS experiments reveal a storage dependent increase in the RBCcm’s bending modulus κ by a factor of 2.8, from 1.9 kBT to 5.3 kBT. MD simulations were conducted in the absence of proteins. The results show that the membrane composition has a small contribution to the increased bending rigidity and suggests additional protein-driven mechanisms.

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MEDUSA: A cloud-based tool for the analysis of X-ray diffuse scattering to obtain the bending modulus from oriented membrane stacks

Himbert,

Gaboo,

Brookes

et al. 2024

PLoS Comput Biol

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An important mechanical property of cells is their membrane bending modulus, κ. Here, we introduce MEDUSA (MEmbrane DiffUse Scattering Analysis), a cloud-based analysis tool to determine the bending modulus, κ, from the analysis of X-ray diffuse scattering. MEDUSA uses GPU (graphics processing unit) accelerated hardware and a parallelized algorithm to run the calculations efficiently in a few seconds. MEDUSA’s graphical user interface allows the user to upload 2-dimensional data collected from different sources, perform background subtraction and distortion corrections, select regions of interest, run the fitting procedure and output the fitted parameters, the membranes’ bending modulus κ, and compressional modulus B.

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Hybrid Erythrocyte Liposomes: Hybrid Erythrocyte Liposomes: Functionalized Red Blood Cell Membranes for Molecule Encapsulation (Adv. Biosys. 3/2020)

Cited by 4 publications

References 0 publications

Erythro-VLPs: Anchoring SARS-CoV-2 spike proteins in erythrocyte liposomes

Erythro-VLPs: Anchoring SARS-CoV-2 spike proteins in erythrocyte liposomes

Blood bank storage of red blood cells increases RBC cytoplasmic membrane order and bending rigidity

MEDUSA: A cloud-based tool for the analysis of X-ray diffuse scattering to obtain the bending modulus from oriented membrane stacks

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