Regulation of the Complement-Mediated Elimination of Red Blood Cells Modified with Biotin and Streptavidin

Muzykantov, Vladimir R.; Murciano, Juan-Carlos; Taylor, Ronald P.; Atochina, Elena N.; Herráez, Ángel

doi:10.1006/abio.1996.0384

Cited by 57 publications

(44 citation statements)

References 15 publications

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“…We then considered coupling neprilysin and the other Aβ degrading enzymes to erythrocytes through a biotin-streptavidin linkage as described by Muzykantov et al [34]. Therefore the effect of biotinylation on peptidase activity was first determined by comparing the activity of the derivatized and underivatized peptidases using synthetic substrates.…”

Section: Resultsmentioning

confidence: 99%

“…In order to couple the biotinylated peptidases to erythrocytes we isolated and biotinylated mouse erythrocytes according to the method of Muzykantov et al [34]. It has previously been observed that there is a limit to which erythrocytes can be biotinylated after which they become unstable [34].…”

Section: Resultsmentioning

confidence: 99%

“…It has previously been observed that there is a limit to which erythrocytes can be biotinylated after which they become unstable [34]. We therefore tested the stability of erythrocytes modified at concentrations of biotin N-hydroxysuccinimide of 25, 50 and 75 mM by reconstituting the biotinylated erythrocytes in fresh mouse serum.…”

Section: Resultsmentioning

confidence: 99%

“…Mouse erythrocytes were biotinylation with biotin-NHS as described by Muzykantov et al [34]. Briefly, fresh mouse erythrocytes were washed 3 times with PBS and incubated with 50 μM biotin-NHS in PBS for 1 h at room temperature.…”

Section: Preparation Of Biotinylated and Erythrocyte-conjugated Peptimentioning

confidence: 99%

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In vitro and in vivo degradation of Aβ peptide by peptidases coupled to erythrocytes

et al. 2007

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It is generally believed that amyloid β peptides (Aβ) are the key mediators of Alzheimer's disease. Therapeutic interventions have been directed toward impairing the synthesis or accelerating the clearance of Aβ. An equilibrium between blood and brain Aβ exists mediated by carriers that transport Aβ across the blood brain barrier. Passive immunotherapy has been shown to be effective in mouse models of AD, where the plasma borne antibody binds plasma Aβ causing an efflux of Aβ from the brain. As an alternative to passive immunotherapy we have considered the use of Aβ-degrading peptidases to lower plasma Aβ levels. Here we compare the ability of three Aβ-degrading peptidases Publisher's Disclaimer: This is a PDF file of an unedited manuscript that has been accepted for publication. As a service to our customers we are providing this early version of the manuscript. The manuscript will undergo copyediting, typesetting, and review of the resulting proof before it is published in its final citable form. Please note that during the production process errors may be discovered which could affect the content, and all legal disclaimers that apply to the journal pertain. NIH Public Access

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Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Section: Preparation Of Biotinylated and Erythrocyte-conjugated Peptimentioning

confidence: 99%

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In vitro and in vivo degradation of Aβ peptide by peptidases coupled to erythrocytes

et al. 2007

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“…The in vitro properties and in vivo circulation of HPG modified RBCs have been investigated in our group recently 8,11 . The partition of RBCs in Dextran500K/PEG8K aqueous two phase system, supplemented with NaCl and sodium phosphate, depends on the erythrocyte surface charge and on surface glycoprotein composition 16 . Depending on the extent of RBC glycocalyx modification with HPGs, modified RBCs tend to partition from the lower dextran to the upper PEG phase.…”

Section: Discussionmentioning

confidence: 99%

Antigens Protected Functional Red Blood Cells By The Membrane Grafting Of Compact Hyperbranched Polyglycerols

Chapanian

Constantinescu

Brooks

et al. 2013

JoVE

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Red blood cell (RBC) transfusion is vital for the treatment of a number of acute and chronic medical problems such as thalassemia major and sickle cell anemia [1][2][3] . Due to the presence of multitude of antigens on the RBC surface (~308 known antigens 4 ), patients in the chronic blood transfusion therapy develop alloantibodies due to the miss match of minor antigens on transfused RBCs 4, 5 . Grafting of hydrophilic polymers such as polyethylene glycol (PEG) and hyperbranched polyglycerol (HPG) forms an exclusion layer on RBC membrane that prevents the interaction of antibodies with surface antigens without affecting the passage of small molecules such as oxygen ,glucose, and ions 3 . At present no method is available for the generation of universal red blood donor cells in part because of the daunting challenge presented by the presence of large number of antigens (protein and carbohydrate based) on the RBC surface and the development of such methods will significantly improve transfusion safety, and dramatically improve the availability and use of RBCs. In this report, the experiments that are used to develop antigen protected functional RBCs by the membrane grafting of HPG and their characterization are presented. HPGs are highly biocompatible compact polymers 6, 7 , and are expected to be located within the cell glycocalyx that surrounds the lipid membrane 8, 9 and mask RBC surface antigens 10,11 . Video LinkThe video component of this article can be found at http://www.jove.com/video/50075/ Protocol A. Hyperbranched Polyglycerol Modification (SS-HPG)1. Place lyophilized HPG 60 kDa (0.5 g, 0.0083 mmol) in a round bottom flask and dry it overnight under vacuum at 90 °C. 2. Refrigerate the flask to room temperature, and dissolve the dried HPG in anhydrous pyridine (3 ml). 3. To functionalize approximately eight hydroxyl groups on HPG with carboxyl groups, add catalytic amount of dimethylaminopyridine (one drop of 5 mg/ml solution in pyridine) to the HPG solution. To this mixture, add succinic anhydride, (0.0067 g, 0.0664 mmol) dissolved in 0.5 ml pyridine drop wise over 10 min. Stir the mixture overnight at room temperature under argon. 4. Precipitate the mixture in 40 ml of cold acetone (4 °C) in a 50 ml centrifuge tube, and centrifuge using a Beckman J2-MC centrifuge at 27,000x g for 15 min. Decant the supernatant, and remove residual acetone by flushing with argon at room temperature. 5. To activate the carboxyl groups with succinimidyl succinate (SS) groups, dissolve the carboxyl -functionalized HPG in 3 ml of anhydrous DMF.Add N-hydroxysuccinimide (0.0077 g, 0.0664 mmol) and N, N'-diisopropylcarbodiimide (0.0084 g, 0.0664 mmol) to the HPG solution and stir the mixture overnight at room temperature under argon. 6. Purify the SS-HPG by precipitation in cold acetone. 7. Remove residual acetone by flushing with argon. 8. Determine the purity of modified HPG and the degree of carboxyl and SS functionalization by proton ( 1 H)-NMR analysis.

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Hybrid Erythrocyte Liposomes: Functionalized Red Blood Cell Membranes for Molecule Encapsulation

et al. 2020

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The modification of erythrocyte membrane properties provides a new tool towards improved drug delivery and biomedical applications. The fabrication of hybrid erythrocyte liposomes is presented by doping red blood cell membranes with synthetic lipid molecules of different classes (PC, PS, PG) and different degrees of saturation (14:0, 16:0–18:1). The respective solubility limits are determined, and material properties of the hybrid liposomes are studied by a combination of X‐ray diffraction, epi‐fluorescent microscopy, dynamic light scattering (DLS), Zeta potential, UV‐vis spectroscopy, and Molecular Dynamics (MD) simulations. Membrane thickness and lipid orientation can be tuned through the addition of phosphatidylcholine lipids. The hybrid membranes can be fluorescently labelled by incorporating Texas‐red DHPE, and their charge modified by incorporating phosphatidylserine and phosphatidylglycerol. By using fluorescein labeled dextran as an example, it is demonstrated that small molecules can be encapsulated into these hybrid liposomes.

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Regulation of the Complement-Mediated Elimination of Red Blood Cells Modified with Biotin and Streptavidin

Cited by 57 publications

References 15 publications

In vitro and in vivo degradation of Aβ peptide by peptidases coupled to erythrocytes

In vitro and in vivo degradation of Aβ peptide by peptidases coupled to erythrocytes

Antigens Protected Functional Red Blood Cells By The Membrane Grafting Of Compact Hyperbranched Polyglycerols

Hybrid Erythrocyte Liposomes: Functionalized Red Blood Cell Membranes for Molecule Encapsulation

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