The Lip-ARM formulation was found to be 10 mg for 100 mg lipids and 10 mL PBS buffer, with an EPC-CHO molar ratio of 4:1. Briefly, The ARM, EPC and CHO were simultaneously dissolved in a 3 mL mixture solution of absolute ethanol and dichloromethane (ratio of volume was 5:1), then this solution was added dropwise slowly to PBS (45 °C, 10 mL, pH = 7.4) at a rate of 0.23 mL/min and continuously stirred on magnetic stirrer at 750 rpm. Finally the formulation was sonicated for 15 min (Scientz-IID, 200 Hz), subsequently extruded through a 200 nm nuclepore hydrophilic membrane using a LiposoFast extruder apparatus (Avestin, Ottawa, Canada) and residual organic solvents were removed by rotating evaporation to obtain Lip-ARM. The erythrocyte membrane (RBCm) was isolated as previously reported [15 (link)], and its protein was determined by BCA assay kit. The RBCm and Lip-ARM were ultrasonicated together and then passed through 200 nm Nuclepore hydrophilic membrane back and forth at least 20 times to prepare the EM-ARM with different protein concentrations (0.3, 0.6, and 0.9 mg mL− 1). Ultimately, the PEM-ARM was finished by a simple sonication and extrusion through a 200 nm Nuclepore hydrophilic membrane after putting appropriate amount of DSPE-PEG2000-CLIPPKF into the EM-ARM system.
Liposofast extruder apparatus
Manufactured by Avestin
Sourced in Canada, United Kingdom
The LiposoFast extruder apparatus is a laboratory instrument designed for the preparation of liposomes and other lipid-based vesicles. The core function of this equipment is to extrude suspensions of lipids through a membrane with defined pore sizes, resulting in the formation of unilamellar vesicles with a narrow size distribution.
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Lab products found in correlation
2 protocols using liposofast extruder apparatus
1
Erythrocyte-Membrane Armored Liposomes
2
Preparation and Characterization of Erythrocyte-Mimicking Liposomes
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RM-PL was prepared by a thin-film hydration and extrusion method in accordance with previously reported protocols with modifications25 (link). Briefly, 2.0 mg of EPC, 1.6 mg of cholesterol and 0.4 mg of mPEG2000-DSPE were dissolved in 3 mL of dichloromethane, and the solvent was then evaporated in a round flask through a rotary evaporator (Rotavapor R100, Buchi, Switzerland) to form a thin film. The lipid film was hydrated with 1.85 mL of double-distilled H2O and 150 μL of RBC membrane suspension or with 2 mL of double-distilled H2O to prepare RM-PL or conventional PEGylated liposomes (PL). Next, the suspension was sonicated in ice-cold water for 4 min using a bath sonicator and then extruded through 0.2 and 0.1 μm polycarbonate membranes (Nuclepore Track-Etched Membranes, Whatman, UK) using a LiposoFast extruder apparatus (Avestin, LF-1, Canada). The resulting RM-PL was stored at 4 °C until use. To prepare RMV of a comparable size, the RBC membrane suspension suspended in 0.2 mmol/L EDTA solution was directly sonicated in ice-cold water for 4 min followed by extrusion through 0.2 and 0.1 μm polycarbonate membranes. For fluorescence microscopy imaging and in vivo particle tracking purposes, DiD-labelled RM-PL was prepared as described above except that 8 μg of DiD was added to the lipids prior to the thin film formation.
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