Preparation method of ultra-small bionic nanoparticles based on erythrocyte membrane

A red blood cell membrane, nanoparticle technology, applied in nanotechnology, nanotechnology, nanomedicine, etc., can solve the problems of product residue, low efficiency, difficult to obtain, etc., and achieve the effect of stable reaction and mild environment.

Pending Publication Date: 2021-08-13
THE SECOND HOSPITAL AFFILIATED TO WENZHOU MEDICAL COLLEGE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This solution uses mechanical extrusion to wrap the erythrocyte membrane vesicles and PLGA, which is inefficient in manufacturing, and there will be a large amount of product residues that are difficult to obtain

Method used

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  • Preparation method of ultra-small bionic nanoparticles based on erythrocyte membrane
  • Preparation method of ultra-small bionic nanoparticles based on erythrocyte membrane
  • Preparation method of ultra-small bionic nanoparticles based on erythrocyte membrane

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0044] Preparation and characterization of PLGA particles

[0045] Prepare PLGA particles by nano-co-precipitation method: Dissolve PLGA (0.67 dL / g) with a viscosity of 50:50 and carboxy-terminal modification in acetone to 7 mg / mL for use. The above 1 mL PLGA acetone solution was quickly added to 3 mL PH=8.0 Tris-HCl deionized water pre-cooled at 4°C, and then the stirring speed was kept at 600 rpm / min until the acetone was completely evaporated. The collected PLGA nanoparticles were prepared and collected for basic physical characterization, including observing the particle morphology with a transmission electron microscope and detecting the hydraulic size and surface potential of the particles with a particle size potentiometer. figure 1 The TEM results show that the size of PLGA nanoparticles is about 30 nanometers; figure 2 The particle size potentiometer test results show that the hydraulic size of PLGA nanoparticles is about 35 nanometers, and the surface potential is ...

Embodiment 2

[0047] Extraction and purification of ICR mouse erythrocyte membrane

[0048] Erythrocyte extraction from ICR mice: Blood was drawn through the cheek and mixed with 10 mM EDTA in PBS at a volume ratio of 1:1. Centrifuge at 3500rpm for 15min, carefully remove the serum and white blood cell layer, and take the red blood cell layer. Red blood cells were added to deionized water at a volume ratio of 1:6, mixed well and incubated in an ice bath for 30 min. Subsequently, a certain volume of 20XPBS was added to adjust the erythrocyte membrane solution to 1X PBS, and then centrifuged at 13800rpm for 10min to remove hemoglobin in the supernatant. The above process was repeated 3 to 4 times until no hemoglobin was detected in the centrifuged supernatant. The supernatant was removed, the red blood cell membrane was resuspended with deionized water, and the membrane protein concentration was measured by BCA method, and finally stored at -80°C. image 3 Represents red blood cells before...

Embodiment 3

[0050] Preparation, characterization and in vitro stability determination of erythrocyte membrane-encapsulated biomimetic nanoparticles (PLGA@RBC)

[0051]Dilute PLGA:RBC to 1.5mL according to 1:2 (w / w), then add it into a 20mL glass bottle, and ultrasonicate in a water bath for 5min to prepare biomimetic nanoparticles wrapped in red blood cell membrane, which is designated as PLGA@RBC. The prepared and collected PLGA@RBC were subjected to basic physical characterization, including the observation of particle morphology by transmission electron microscopy and the detection of the hydraulic size and surface potential of the particles by a particle size potentiometer. image 3 The TEM results show that PLGA@RBC presents a core-shell structure with a size of about 50 nanometers; Figure 4 The particle size potentiometer test results show that the hydraulic size of PLGA@RBC is about 60 nanometers, and the surface potential is about -27mV. Figure 5 It was shown that PLGA@RBC has ...

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Abstract

The invention discloses a preparation method of ultra-small bionic nanoparticles based on an erythrocyte membrane. The preparation method comprises the following steps: step 1, preparing a PLGA acetone solution; step 2, the PLGA acetone solution is rapidly added into a pre-cooled Tris-HCl deionized water; step 3, stirring is kept till acetone is completely volatilized, and PLGA nanoparticles are obtained; step 4, obtaining an erythrocyte membrane, and mixing the erythrocyte membrane with the PLGA nanoparticles to obtain a mixed solution; and step 5, carrying out water bath ultrasonic treatment on the mixed solution to obtain the bionic nanoparticles wrapped by the erythrocyte membrane. Compared with the prior art, the preparation method has the advantages that the specification is smaller, the biological barrier penetration effect is higher, the application potential is wider, and compared with the prior art, the synthesis efficiency is higher.

Description

technical field [0001] The invention relates to the technical field of biopharmaceuticals, in particular to a method for preparing ultra-small bionic nanoparticles based on red blood cell membranes. Background technique [0002] In recent years, biomimetic nanomedicine technology has been widely used in the fields of drug delivery, biological detoxification, and vaccine preparation. Among them, biomimetic nanomaterials based on red blood cell membranes are the most studied and widely used, which is mainly due to the fact that red blood cells are derived from The body itself, so the biomimetic nanomaterials wrapped in red blood cell membranes have excellent biocompatibility and low immunogenicity, which can effectively avoid the capture of the monocyte-macrophage system and the reticuloendothelial system, and maximize the lifespan of nanoparticles. Blood circulation time in the body. On this basis, erythrocyte membrane-coated particles can deliver a variety of drugs to targe...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K9/51A61K47/46A61K47/34B82Y5/00B82Y40/00
CPCA61K9/5068A61K9/5073A61K9/5031B82Y5/00B82Y40/00
Inventor 陈一杰陈梦纯陈赛玲王芳杨雪薇
Owner THE SECOND HOSPITAL AFFILIATED TO WENZHOU MEDICAL COLLEGE
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