Lipidosome-protected nano-gold gene vector and preparation method thereof

A gene carrier and nano-gold technology, applied in the field of gene carriers, can solve the problems of inability to release the target gene, inability to express the target gene, slow release of the target gene, etc. Effect

Active Publication Date: 2014-04-16
CHANGCHUN INST OF APPLIED CHEMISTRY - CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, gold nanoparticles protected by DDAB cannot achieve the release of the target gene in the cell because of its strong interaction with the target gene, and cannot make the target gene expressed in the cell (Journal of Controlled Release2008,129,128-134)
DODAB protected gold nanoparticles can achieve the expressi

Method used

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  • Lipidosome-protected nano-gold gene vector and preparation method thereof
  • Lipidosome-protected nano-gold gene vector and preparation method thereof
  • Lipidosome-protected nano-gold gene vector and preparation method thereof

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preparation example Construction

[0041] The invention discloses a preparation method of a liposome-protected nano-gold gene carrier, which comprises the following steps:

[0042](A) Dissolving di-octadecyldimethylammonium bromide and dioleoylphosphatidylethanolamine in an organic solvent, then removing the organic solvent, dissolving in water, and ultrasonically treating to obtain a phospholipid vesicle solution;

[0043] The molar ratio of dioctadecyldimethylammonium bromide to dioleoylphosphatidylethanolamine is 1:10~10:1;

[0044] (B) After the phospholipid vesicle solution is mixed with a compound containing gold ions, a reducing agent is added to react to obtain a liposome-protected nano-gold gene carrier.

[0045] In the present invention, dioctadecyldimethylammonium bromide and dioleoylphosphatidylethanolamine are dissolved in an organic solvent first, then the organic solvent is removed and dissolved in water, and the phospholipid vesicle solution is obtained by ultrasonication. The molar ratio of di...

Embodiment 1

[0050] Dissolve 0.0032g of DODAB and 0.0018g of DOPE in 10mL of chloroform, remove most of the chloroform with nitrogen flow, and then vacuum dry for 2 hours to remove traces of chloroform. After adding 7 mL of aqueous solution, sonicate until clarified to prepare DODAB / DOPE vesicle solution. Then, 7 μl of 0.1M chloroauric acid solution was added to the prepared vesicle solution and stirred to mix. After mixing, 8 μl of 0.4 M sodium borohydride solution was added. DODAB / DOPE-protected gold nanoparticles were obtained after stirring for 1 hour.

[0051] Remove 500 µl and record the UV-Vis absorption spectrum at room temperature, see figure 1 , figure 1 It is the ultraviolet absorption spectrum of the nano gold gene carrier that the liposome protection that embodiment 1 obtains. figure 2 Electron micrograph of the nano-gold gene carrier protected by the liposome prepared in Example 1. Depend on figure 1 and figure 2 It can be seen that the present invention has obtained...

Embodiment 2

[0054] Dissolve 0.0032g of DODAB and 0.0037g of DOPE in 10mL of chloroform, remove most of the chloroform with nitrogen flow, and then vacuum dry for 2 hours to remove traces of chloroform. After adding 10 mL of aqueous solution, sonicate until clarified to prepare a DODAB / DOPE vesicle solution. Then, 10 microliters of 0.1M chloroauric acid solution was added to the prepared vesicle solution and stirred to mix. After mixing, 12 microliters of 0.4M sodium borohydride solution was added. DODAB / DOPE-protected gold nanoparticles were obtained after stirring for 1 hour.

[0055] Take out 500 µl and record the UV-Vis absorption spectrum at room temperature. see Figure 4 , Figure 4 The UV absorption spectrum of the liposome-protected nano-gold gene carrier obtained in Example 2. Figure 5 Electron micrograph of the nano-gold gene carrier protected by the liposome prepared in Example 2. Depend on Figure 4 and Figure 5 It can be seen that the present invention has obtained ...

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Abstract

The invention provides a preparation method of a lipidosome-protected nano-gold gene vector. The preparation method comprises the following steps: dissolving dimethyldioctadecylammonium bromide and dioleoyl phosphatidyl ethanolamine into an organic solvent, removing the organic solvent, dissolving into water, and performing ultrasonic treatment to obtain a phospholipid vesicle solution, wherein the molar ratio of the dimethyldioctadecylammonium bromide to the dioleoyl phosphatidyl ethanolamine is 1:10-10:1; and mixing the phospholipid vesicle solution with a compound containing gold ions, adding a reducing agent, and reacting to obtain the lipidosome-protected nano-gold gene vector. After being compounded with genes, the lipidosome-protected nano-gold gene vector is high in transfection efficiency and high in gene release speed in a specific environment; the lipidosome-protected nano-gold gene vector is high in stability in serum and is not aggregated, and a transfection process is simplified since serum in a culture solution does not need to be removed in an in-vitro transfection process; the carrying amount of DNAs (Desoxvribose Nucleic Acids) is high, the using quantity of the DNAs is reduced, and the immunogenicity of exogenous DNAs to cytotoxicity and organisms are lowered.

Description

technical field [0001] The invention relates to the field of gene carriers, in particular to a liposome-protected nano-gold gene carrier and a method thereof. Background technique [0002] Gene therapy is an emerging gene-level treatment method, which can treat single-gene genetic diseases, polygenic diseases, cancer, infectious diseases (such as AIDS), cardiovascular diseases, etc. Gene therapy is different from conventional treatments: in general, the treatment of diseases is aimed at various symptoms caused by gene abnormalities, while gene therapy is aimed at the root of the disease - the abnormal gene itself, so it is a radical cure for the disease. A new method with great medical value. In November 1990, the NIH in the United States conducted the first clinical trial of human gene therapy, transferring the adenosine deaminase (ADA) gene to the patient's white blood cells through retroviruses. Eight months later, the ADA level in the child's body reached 25% of the no...

Claims

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Application Information

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IPC IPC(8): A61K48/00A61K47/24A61K47/02
Inventor 李丹杜宝吉汪劲汪尔康
Owner CHANGCHUN INST OF APPLIED CHEMISTRY - CHINESE ACAD OF SCI
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