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Nanogold gene carrier protected by liposome 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, the inability to express the target gene, and the slow release of the target gene, and achieve the goal of improving transfection efficiency, promoting release, and high loading capacity. Effect

Active Publication Date: 2017-06-06
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 expression of the target gene, and its transfection efficiency is 4 times higher than that of DODAB, but the transfection efficiency is still low, the release of the target gene in the cell is slow, and the expression of the target gene in the cell is required. Disadvantages such as long time (Biomaterials2008, 29, 3617-3624)

Method used

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  • Nanogold gene carrier protected by liposome and preparation method thereof
  • Nanogold gene carrier protected by liposome and preparation method thereof
  • Nanogold gene carrier protected by liposome and preparation method thereof

Examples

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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 liposome-protected nano-gold gene carrier: dissolving dioctadecyldimethylammonium bromide and dioleoylphosphatidylethanolamine in an organic solvent, and then removing the organic solvent After dissolving in water, ultrasonically obtain the phospholipid vesicle solution; the molar ratio of the dioctadecyldimethylammonium bromide and dioleoylphosphatidylethanolamine is 1:10~10:1; the phospholipid vesicle After the capsule solution is mixed with the compound containing gold ions, a reducing agent is added to react to obtain the nano-gold gene carrier protected by the liposome. After the liposome-protected nano-gold gene carrier of the present invention is combined with the gene, the transfection efficiency is high, and the gene release speed is fast in a specific environment; it has good stability in serum, no aggregation phenomenon, and does not need to be cultured during the in vitro transfection process. The serum in the liquid is removed, which simplifies the transfection process; the DNA load is high, which reduces the amount of DNA used, and reduces the cytotoxicity and immunogenicity of exogenous DNA to the body.

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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Patent Type & Authority Patents(China)
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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