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Application of fullerene derivative to prepare gene transmission vector

A technology of fullerene derivatives and gene transfer, which is applied in the field of biotechnology and vaccines, can solve the problems of poor water solubility of fullerenes, achieve the effect of improving defense capabilities, improving cellular immunity and humoral immunity, and good safety

Active Publication Date: 2014-03-12
THE NAT CENT FOR NANOSCI & TECH NCNST OF CHINA +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Therefore the purpose of the present invention is to provide a kind of fullerene derivative, introduces hydrophilic group on its surface, as hydroxyl (-OH), carboxyl (-COOH) etc., can solve the problem that fullerene water solubility is poor, for its Application lays the groundwork

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  • Application of fullerene derivative to prepare gene transmission vector
  • Application of fullerene derivative to prepare gene transmission vector
  • Application of fullerene derivative to prepare gene transmission vector

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] Example 1 Fullerenol C 60 (OH) 20 Synthesis

[0036] First, add 1ml of 40% tetrabutylammonium hydroxide solution (TBAH) and 2ml of sodium hydroxide solution (2.22g / ml) to 30ml of C 60(Sigma company) in toluene solution (1.5mg / ml), react 24h under vigorous stirring; Discard organic phase, and remove aqueous phase with vacuum rotary evaporator (RV06-ML 1-B, Germany); Wash sample with 50ml methanol, Methanol was removed by vacuum evaporation, and washed repeatedly for 3-5 times; after that, 10ml of deionized water was added to the sample, and the stirring was continued until the solution was transparent reddish-brown, then 20ml of deionized water was added, and the Sephadex G-25 chromatographic column was used to (Amersham Company, UK) purified the sample, and finally vacuum-dried to obtain fullerenol nanoparticles. Fullerenols with different numbers of hydroxyl groups can be obtained by adjusting the amount of sodium hydroxide added.

[0037] Utilize atomic ...

Embodiment 2

[0042] Example 2 Effect of Fullererol on Vitality of HEK293 Cells (ATCC )

[0043] 1. Preparation of complete medium containing fullerenol:

[0044] First, take an appropriate amount of fullererol obtained in Example 1, and prepare a 2 mM aqueous solution with sterile deionized water; then use complete DMEM (Dulbecco's modification of Eagle's medium) medium (containing 10% fetal bovine serum, Gibco Company) diluted fullererol aqueous solution to 500 μM, and carried out gradient dilution with complete medium to obtain a series of solutions with concentrations of 500, 100, 20, 4, 0.8, and 0.16 μM for future use.

[0045] 2. The effect of fullerenol on the viability of HEK293 cells:

[0046] First, HEK293 cells were digested with 0.25% trypsin, and after the digestion was terminated, centrifugation was performed at 1000 rpm for 5 min. Single-cell suspension was prepared in DMEM medium containing 10% fetal bovine serum (FBS), and 5 × 10 4 / ml concentration was inoculated in 9...

Embodiment 3

[0048] Example 3 Effect of fullererol on the viability of NIH3T3 cells (ATCC)

[0049] With reference to the method described in Example 2, investigate the effect of fullerenol nanoparticles on the viability of NIH3T3 cells, the results are shown in Figure 6 , similar to the results in Example 2, compared with the negative control group, the cell viability did not change significantly after the cells were treated with fullerenol nanoparticles for 48 hours, even in the highest dose group, the cell viability was still higher than 90%. Figure 7 Indicates the effect of fullerenol on the morphology of NIH3T3 cells, Figure 7 A is the normal control group, Figure 7 B represents the picture of the cells treated for 48 hours when the working concentration of fullerenol was 500 μM. It can be seen that the morphology of the cells in the test group is the same as that in the normal control group, showing a long spindle shape. All these indicate that fullerenol has good biocompatib...

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Abstract

The invention provides a fullerene derivative, which is a fullerol shown by a general formula C60OxHy (y is more than 10 and less than or equal to x, and x is more than or equal to y and less than 50) or a fullerene carboxyl derivative shown by a general formula C60(C(COOH)2)n (n is an integral number of between 1 and 4), wherein the diameter range of nano-particles of the fullerene derivative in a solvent is between 1 and 400nm. The invention also provides application of the vaccine adjuvant of the fullerene derivative in the field of vaccines, and the vaccine adjuvant of the fullerene derivative has high safety, can remarkably improve the immunogenicity of the vaccine through different immune ways, and has great application prospect.

Description

technical field [0001] The present invention relates to a kind of fullerene derivative, specifically, relate to fullerene alcohol particle C 60 o x h y (10<y≤x<50) and fullerene carboxyl derivatives C 60 (C(COOH) 2 ) n (n is an integer of 1-4) and the vaccine adjuvant and vaccine preparation of the fullerene derivative and its application in the field of vaccines belong to the fields of biotechnology and vaccines. Background technique [0002] Vaccines play an extremely important role in the prevention and treatment of major diseases, especially infectious diseases. In the process of vaccine research and development, safe and effective vaccines are mainly obtained through design optimization of antigens and vaccine carriers or adjuvants. At present, the only clinically approved adjuvant is aluminum adjuvant, which can better promote the body's humoral immunity level and play an important role in the prevention of diseases. However, aluminum adjuvants inhibit the...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K39/39A61K39/12A61K39/21A61K39/29A61K48/00C12N15/63A61P31/18A61P31/20
Inventor 陈春英邵一鸣许利耕郝彦玲刘颖刘野
Owner THE NAT CENT FOR NANOSCI & TECH NCNST OF CHINA
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