PEGylation low molecular mass PEI derivative, preparation method and application thereof, and its compound

A low molecular weight, polyethylene glycol succinimide carbonate technology, applied in the biological field, to achieve low cytotoxicity, simple structure, and low surface charge density

Active Publication Date: 2015-09-16
SHANGHAI JIAO TONG UNIV
View PDF2 Cites 2 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, these materials have cer

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
  • PEGylation low molecular mass PEI derivative, preparation method and application thereof, and its compound
  • PEGylation low molecular mass PEI derivative, preparation method and application thereof, and its compound
  • PEGylation low molecular mass PEI derivative, preparation method and application thereof, and its compound

Examples

Experimental program
Comparison scheme
Effect test

Example Embodiment

[0056] Example 1. Preparation of polyethylene glycol (PEG) polyethyleneimine (PEI) derivative (PEG-Et 1:1)

[0057] Such as figure 1 As shown, the schematic diagram of the preparation synthetic route of the PEGylated polyethyleneimine (PEI) derivative of the present invention includes the following steps:

[0058] (a) Dissolve PEI-Et and mPEG-SC with sodium bicarbonate respectively, and make 2mL and 5ml solutions respectively; combine PEI-Et and monomethoxy polyethylene glycol succinimide carbonate (mPEG- SC) The molar ratio is 1:1 and added to 0.1M sodium bicarbonate solution.

[0059] (b) At room temperature, add the sodium bicarbonate solution of monomethoxy polyethylene glycol succinimide carbonate dropwise to the PEI-Et sodium bicarbonate reaction system, and stir for 4 hours;

[0060] (c) Separation and purification. The product is placed in a dialysis bag with a molecular weight cut-off of 3500Da for 48 hours (any value from 24 to 48 hours). After the dialysis is completed, the...

Example Embodiment

[0071] Example 2. Preparation of PEG-Et 1:1 and plasmid synthesis complex (Polyplex)

[0072] Weigh a quantitative polymer (PEG-Et 1:1) polymer, add ultrapure water to prepare a 2mg / mL solution, then filter with a 0.45μm sterile filter, and dilute the plasmid concentration to 20μg / mL;

[0073] Configure the complex solution with different mass ratios to keep the concentration of the plasmid solution unchanged, and then dilute the concentration of the polymer solution according to the mass ratio of PEG-Et 1:1 to the plasmid to keep the diluted polymer solution and plasmid solution The volume is equal, and finally the polymer solution is quickly added to the plasmid solution and mixed, and incubated at room temperature for 30 to 60 minutes, so that a series of mass ratio complexes can be obtained, which can be used for further physical and chemical properties determination.

[0074] PEG-Et 1:1 and plasmid synthesis complex (Polyplex) particle size measurement

[0075] The sample size fo...

Example Embodiment

[0083] Example 3 Cell transfection experiment of PEG-Et 1:1 and plasmid synthesis complex (Polyplex)

[0084] In a 48-well cell culture plate, add 500μL of cell suspension (MCF-7, CT-26 or HeLa) with a density of 5.0~10×10 4 / mL, incubate overnight. When transfecting a 48-well plate, configure the polymer to a 2mg / mL solution, and filter it with a 0.45μm filter membrane aseptically, dilute it to the required ratio according to the set mass ratio of the sample to be tested to the plasmid, and then polymerize Add the substance solution to the plasmid solution, mix quickly, and incubate for 30 min. In this way, the volume of the complex added to each well is 50 μL, which is one-tenth of the total volume (500 μL), which meets the regulations. Each mass ratio makes 3 replicate holes. Positive control group PEI 25kDa, with its best mass ratio of 2, make 3 control wells each. During this period of incubation, remove the cells from the incubator, remove the medium with serum, and use 2...

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

No PUM Login to view more

Abstract

The invention discloses a PEGylation low molecular mass PEI derivative, a preparation method and an application thereof, and its compound. Compared with prior art, the prepared PEGylation PEI derivative has high transfection activity and little cytotoxicity, has good biological activity in different cells, and is high-efficiency and low-toxicity gene substance carrier used for conveying gene substance.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a polyethylene glycol (PEG) PEI derivative, a preparation method, an application and a compound thereof. Background technique [0002] Gene therapy is the introduction of exogenous genetic material (DNA or RNA) into cells to promote or inhibit the expression of specific proteins, or replace or repair problematic genes, so as to achieve the purpose of disease treatment. In the body, DNA and RNA can be rapidly degraded by DNase and RNase and excreted through the kidneys; and DNA and RNA have large molecular weights and negative charges, so naked DNA and RNA cannot pass through the cell membrane and enter the cell. Therefore, the key to applying gene therapy to human body is to find a safe and effective gene carrier. [0003] Commonly used gene delivery vectors can be divided into recombinant viral vectors and non-viral vectors. In clinical applications, the safety of vira...

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
IPC IPC(8): C08G81/00C12N15/87C12N15/85
Inventor 邱明丰苏靖马琳孙雅楠张彪陆平盛净
Owner SHANGHAI JIAO TONG UNIV
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products