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Preparation method of nano aluminum adjuvant/ autologous tumor vaccine

A tumor vaccine and nano-aluminum technology, applied in the field of nano-aluminum adjuvant/autologous tumor vaccine and its preparation, can solve problems such as imperfection, and achieve the effects of uniform distribution, uniform particle size and small particle size

Inactive Publication Date: 2014-07-30
SOUTHEAST UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, there are still imperfections in the use of nanoparticles as adjuvants, such as the combination of nanoparticles and vaccine antigens, the effect of the combination ratio of nanoparticles and vaccines on the protective immunity of vaccines, and the stability of antigens during encapsulation and microsphere storage. , and the hydration of the microspheres needs to be resolved before the controlled release of the vaccine

Method used

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  • Preparation method of nano aluminum adjuvant/ autologous tumor vaccine
  • Preparation method of nano aluminum adjuvant/ autologous tumor vaccine
  • Preparation method of nano aluminum adjuvant/ autologous tumor vaccine

Examples

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preparation example Construction

[0027] The method for preparing nano aluminum adjuvant / autologous tumor vaccine of the present invention, comprises the following steps:

[0028] 1) Preparation of nano aluminum adjuvant:

[0029] Mix benzalkonium bromide, n-octanol, and cyclohexane at a high speed at a mass ratio of 1:1:1 and stir evenly. The resulting mixed solution is placed on a magnetic stirrer, and 3 The mass ratio is 1:1.3~1.5, and the latter is added into the mixed solution and stirred for more than 20 minutes. Then add ammonia water dropwise to the mixed solution at a rate of 20-30 drops / min, keep the pH of the reaction system > 10 and react for 2 hours to obtain an emulsion. After the reaction, add acetone to the emulsion until the emulsion becomes clear, centrifuge at a speed of >500 rpm, discard the supernatant, and obtain a precipitate. Finally, the precipitate was centrifuged and washed three times with double distilled water and ethanol respectively, and then dried in a drying oven to obtain t...

Embodiment 1

[0035] 1) Preparation of nano aluminum adjuvant:

[0036] Weigh benzalkonium bromide, n-octanol, and cyclohexane according to a mass ratio of 1:1:1, stir them at high speed, pour them into a conical flask, place them on a magnetic stirrer, and mix n-octanol and AlCl 3 The mass ratio is 1:1.3, and the latter is added to the aforementioned solution and stirred for more than 20 minutes. Then add ammonia water dropwise at a rate of 20 drops / min, keep the pH of the reaction system at 10 and react for 2 hours to obtain an emulsion. After the reaction, add acetone until the emulsion becomes clear, centrifuge at a speed of 500 rpm, and discard the supernatant. Then the precipitate was centrifuged and washed three times with double distilled water and ethanol respectively to obtain a precipitate. The above precipitate is placed in a drying oven and dried overnight to obtain the nano-aluminum adjuvant.

[0037] 2) Preparation of nano aluminum adjuvant / autologous tumor vaccine:

[00...

Embodiment 2

[0040] Embodiment 2: The basic process is the same as in Embodiment 1, and the difference with Embodiment 1 is:

[0041] Step 1) n-octanol and AlCl 3 The mass ratio is 1:1.5, then stir for 20 minutes, and add ammonia water at a rate of 30 drops / min and stir. The pH of the reaction system was 14. Centrifuge after adding acetone, the centrifugal speed is 1000rpm.

[0042] In step 2), the cell suspension concentration is 3×10 8 After the cells were treated in a water bath at 65°C for 30 minutes, they were co-incubated with 4 mg / ml nano-aluminum adjuvant and common aluminum adjuvant at 4°C for 7 days.

[0043] The other proportional relationships of this embodiment are the same as those of Embodiment 1.

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Abstract

The invention discloses a preparation method of a nano aluminum adjuvant / autologous tumor vaccine. The preparation method comprises the following steps: firstly, preparing a nano aluminum adjuvant by use of a micro-emulsion method, namely after adding and stirring benzalkonium bromide, n-octyl alcohol and cyclohexane in equal proportion at a high speed, adding a proper amount of AlCl3 liquor and then stirring; slowly dropwise adding ammonia water, reacting for 2 hours by keeping PH of a reaction system greater than 10, then adding acetone, performing demulsification and centrifugation, centrifuging and washing precipitates for three times, drying the centrifuged precipitates in a drying oven and staying overnight to obtain a fluffy nano Al(OH)3 adjuvant; then, co-incubating the H22 liver cancer cell suspension liquid with the nano aluminum adjuvant to obtain the nano aluminum adjuvant / autologous tumor vaccine. The nano aluminum adjuvant / autologous tumor vaccine prepared by the preparation method disclosed by the invention is small in particle size, good in dispersion, better in stability and good in biocompatibility. Animal experiment results show that the anti-tumor effect of the nano aluminum adjuvant / autologous tumor vaccine is obviously superior to that of the common aluminum adjuvant.

Description

technical field [0001] The invention belongs to the field of anti-tumor autologous vaccines, and relates to a nano-aluminum adjuvant / autologous tumor vaccine for treating cancer and a preparation method thereof. Background technique [0002] So far, tumor is still a persistent disease that threatens human health and life. As a new mode of treatment, tumor biotherapy has become an irreplaceable and important part of comprehensive tumor treatment, which mainly includes two aspects: immunotherapy and gene therapy. The purpose of tumor immunotherapy is to stimulate and mobilize the immune system, enhance the anti-tumor immunity of the tumor microenvironment, and thus control and kill tumor cells. From the analysis of the current international research trends, it is speculated from the perspective of practical application that immunotherapy will have broad research prospects. The research on immunotherapy includes the development of vaccines and the development of new immune ad...

Claims

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Application Information

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IPC IPC(8): A61K39/39A61K39/00A61P35/00
Inventor 张东生刘慧杜益群张皓
Owner SOUTHEAST UNIV
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