Multifunctional polypeptide/liposome/hyaluronic acid assembled viroid nucleic acid carrier
A nucleic acid carrier and hyaluronic acid technology, applied in the fields of genetic material components, recombinant DNA technology, and other methods of inserting foreign genetic materials, etc., can solve the problems of complicated problems, no new advantages in shielding charges, and less than ideal effects. , to avoid toxicity and break through the long-term circulation in the body
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Problems solved by technology
Method used
Image
Examples
Example Embodiment
[0048] The present invention relates to a preparation method of the above-mentioned nucleic acid carrier, comprising the following steps:
[0049] a. Preparation of cationic liposomes by thin film dispersion method: using cationic lipids ((2,3-dioleyloxypropyl)trimethylammonium chloride (DOTAP)) and neutral lipids (1,2-dioleyloxypropyl) A chloroform solution with a mass ratio of oleoyl-sn-glycerol-3-phosphoethanolamine (DOPE)) was rotary evaporated to remove the solvent, then hydrated overnight, and sonicated for 30-60 minutes.
[0050] b. Preparation of nanocarriers: use the above-prepared liposomes to dilute to a certain concentration, mix them with a certain concentration of polypeptides at a mass ratio of 0.75:4 and 1:4, respectively, and add them to DNA and siRNA. Nanoparticles with positive charges on the surface were prepared, wherein the mass ratio of liposome, polypeptide and nucleic acid of the two types of nanoparticles was 0.75:4:1 and 1:4:1, and incubated at room ...
Example Embodiment
[0052] Embodiment 1, the preparation method of Q-complexes
[0053] like figure 1 As shown, the system for measuring physicochemical properties of Q-complexes was performed in aqueous solution. After mixing the above cationic liposomes and polypeptides in different mass ratios, add them into the nucleic acid aqueous solution and continue to mix them evenly. After incubating at room temperature for 30 minutes, polyanions in different proportions were added to the mixture and mixed well and allowed to stand for 15 minutes. Serum-free medium can be used in the carrier system for cell experiments instead of ultrapure water.
Example Embodiment
[0054] Example 2. Gel electrophoresis of Q-complexes
[0055] Prepare a mass ratio of 1.0% agarose solution, heat and dissolve in a microwave oven, take 40 mL of the solution, pour it into a special EB-contaminated beaker, add about 4 μL of EB solution, stir well, pour it into the mold, insert a comb, about 30min After that, the gel will solidify, add an appropriate amount of TAE buffer to the electrophoresis tank, put the agarose gel into the electrophoresis tank and wait for sample loading. Then, complex solutions with different mass ratios were prepared and incubated at room temperature for 30 min. The marker for loading is a 1000-10000kb plasmid maker. When loading, first take 1 μL of loading buffer, add 5 μL of sample sample, mix well, and add it to the gel well. Add a voltage of 120 volts, the blue bromophenol blue will quickly move to the bottom of the gel, about 20 minutes later, take a picture with a UV gel imaging system. Gel electrophoresis results of complexes ...
PUM
Property | Measurement | Unit |
---|---|---|
Viscosity | aaaaa | aaaaa |
Particle size | aaaaa | aaaaa |
Particle size | aaaaa | aaaaa |
Abstract
Description
Claims
Application Information
- R&D Engineer
- R&D Manager
- IP Professional
- Industry Leading Data Capabilities
- Powerful AI technology
- Patent DNA Extraction
Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic.
© 2023 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap