Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

A kind of endoplasmic reticulum membrane fusion liposome and its preparation method and application

An endoplasmic reticulum membrane and liposome technology, applied in the biological field, can solve the problems of complex chemical reaction, high exploration cost, uncertain biocompatibility, etc., and achieve the effect of reducing Zeta potential and facilitating industrial transformation.

Active Publication Date: 2022-07-26
SHANDONG UNIV
View PDF0 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the discovery and design of these targeting molecules depend on the comprehensive analysis of target proteins located on the endoplasmic reticulum, and the cost of exploration is too high; well-designed synthetic materials are not conducive to clinical practice due to their uncertain biocompatibility and complex chemical reactions. convert

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • A kind of endoplasmic reticulum membrane fusion liposome and its preparation method and application
  • A kind of endoplasmic reticulum membrane fusion liposome and its preparation method and application
  • A kind of endoplasmic reticulum membrane fusion liposome and its preparation method and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0039] Example 1: Extraction of endoplasmic reticulum membrane from B16F10 cells

[0040] Prepare 10 T75 flasks of B16F10 cells (count 5*10 7 ), washed twice with PBS buffer, each bottle was digested with 3 mL of trypsin ethylenediaminetetraacetic acid hydrochloride for 2 min, and DMEM complete medium was added to terminate the digestion, and the cell digest was collected and centrifuged at 300 g for 5 min. Discard the supernatant and keep the cell pellet. Add 2 mL of pre-chilled grinding buffer (sigma) and protease inhibitors (Biyuntian), transfer to a pre-chilled DOUNCE homogenizer, and homogenize for 50 times on ice until more than 80% cell lysis is observed under the microscope. The homogenate was transferred to a centrifuge tube, centrifuged at 1000 g at 4°C to remove nuclei and cell debris, and the supernatant was collected. The supernatant was then centrifuged at 12,000 g for 30 min at 4°C, and the supernatant was collected. Use an ultracentrifuge and matching tube t...

Embodiment 2

[0042] Example 2: Preparation of Doxorubicin (DOX)-loaded Liposomes by Ammonium Sulfate Gradient Method and Preparation of EM / DOX Lip by Ultrasonic Mixing

[0043] First, DOPC, DOPE, and CHEMS were dissolved in 3 ml of chloroform in a ratio of 1:1:0.5, and then placed in a rotary evaporator for rotary decompression evaporation. The temperature of the water bath was 25 °C and the speed was 70 rpm / min. After forming a transparent and uniform lipid film on the inner wall, it was vacuum dried overnight to completely remove residual solvent. Add 5 mL of ammonium sulfate solution (250 mmol, pH=5.5) preheated to 40 °C and ultrasonically in a water bath to obtain a liposome solution with light blue opalescence. Then, the obtained solution was put into the treated dialysis bag for dialysis for 48 hours, free ammonium sulfate was removed, and the solution was diluted to 1 mg / mL liposome solution.

[0044] Then, 0.1 mg of free doxorubicin solution was added per ml to the obtained soluti...

Embodiment 3

[0049] Example 3: Extraction of endoplasmic reticulum membrane from DC2.4 cells

[0050] Prepare 8 T75 flasks of cells (count 4*10 7), washed twice with PBS buffer, digested with trypsin ethylenediaminetetraacetic acid hydrochloride for 3 min, added complete medium to terminate the digestion, and collected the cell digestion solution by centrifugation at 200 g for 10 min. Discard the supernatant and keep the cell pellet. Add 1 mL of pre-cooled grinding buffer (sigma) and protease inhibitors (Biyuntian), transfer to a pre-cooled DOUNCE homogenizer, and homogenize for 20 times on ice until most cells are lysed under a microscope. The homogenate was transferred to a centrifuge tube, centrifuged at 1000 g at 4°C to remove nuclei and cell debris, and the supernatant was collected. The supernatant was then centrifuged at 12,000 g for 15 min at 4°C, and the supernatant was collected. Centrifuge again at 4°C and 150,000g for 90min using an ultracentrifuge and matching tube, careful...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

PropertyMeasurementUnit
particle diameteraaaaaaaaaa
Login to View More

Abstract

The invention relates to an endoplasmic reticulum membrane fusion liposome, a preparation method and application thereof, and belongs to the field of biotechnology. The steps are: (1) extraction of endoplasmic reticulum membrane; (2) preparation and characterization of liposomes; (3) preparation and characterization of endoplasmic reticulum membrane fusion liposomes. The particle size of the liposome prepared by the film dispersion method is controlled at 103.03±7.99nm, the particle size distribution is narrow, the PDI=0.149±0.02, and the zeta potential is 41.8±2.71mV. After fusion with endoplasmic reticulum membrane, the final endoplasmic reticulum membrane fusion liposome (EM / Lip) particle size was controlled at 124.27±4.21nm, PDI=0.19±0.03, and zeta potential was 14.27±2.50mV. In the present invention, most of the EM / Lip is located in the endoplasmic reticulum after entering the cell, and rarely enters the lysosome, thereby avoiding the degradation of the drug in the lysosome and realizing the targeting function of the endoplasmic reticulum. The endoplasmic reticulum membrane fusion liposome obtained by the invention can be used in the endoplasmic reticulum targeting function of immune cells and tumor cells, and the three-dimensional image technology of laser confocal is also well applied to the research on the distribution of nanometer preparations at the subcellular organelle level.

Description

technical field [0001] The invention relates to an endoplasmic reticulum membrane fusion liposome, a preparation method and application thereof, and belongs to the field of biotechnology. Background technique [0002] With the development of precision medicine, the development of suitable drugs and drug delivery systems based on key biological macromolecules such as genes and proteins for the occurrence and development of the disease itself has become the latest research hotspot. And most of the key biomolecules associated with disease are located in cells. Only precise targeting can achieve precise life regulation. An attractive strategy to improve the therapeutic index of drugs is to not only carry therapeutic agents to target tissues and target cells, but also control the precise targeting of drugs to subcellular structures. Achieve precise drug distribution and release from intracellular targets. However, most studies on drug delivery systems to date have struggled to ...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
Patent Type & Authority Patents(China)
IPC IPC(8): A61K9/127A61K45/00A61K31/704A61K31/7088A61K47/46A61P35/00C12N5/071C12N5/09
CPCA61K9/1271A61K9/1277A61K45/00A61K31/704A61K31/7088A61K47/46A61P35/00C12N5/0693C12N5/0626C12N2509/00C12N2509/10
Inventor 林贵梅傅相蕾邱胜男
Owner SHANDONG UNIV
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com