High-efficiency CpG preparation, preparation method thereof and application

A preparation and high-efficiency technology, applied in the direction of medical preparations containing active ingredients, pharmaceutical formulas, oil/fat/wax non-active ingredients, etc., can solve the problems of decreased drug efficacy, weak stability, harsh storage conditions, etc., to achieve Effects of reduced usage, reduced usage, and low production cost

Active Publication Date: 2014-05-28
SOUTH CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] However, at present, there is no vaccine immunization product that directly applies CpG to animals. The main reason is that its stability is not strong, and it is easy to degrade when stored for a long time, resulting in a decrease in drug efficacy; the storage conditions are relatively harsh, and it is required to be kept at -20°C.

Method used

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  • High-efficiency CpG preparation, preparation method thereof and application
  • High-efficiency CpG preparation, preparation method thereof and application
  • High-efficiency CpG preparation, preparation method thereof and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] The preparation of embodiment 1 CpG preparation

[0031] Prepare raw materials according to the following ratios:

[0032]

[0033] The PBS composition is: by weight percentage, 0.6% NaCl, 0.01% KCl, 0.0124% Na 2 HPO 4 , 0.014% KH 2 PO 4 Soluble in water; damp heat autoclave: 115°C, 15min.

[0034] (1) After CpG ODN was synthesized by TAKARA Company, it was prepared into 20 μg / ml with sterile PBS solution and stored at -15°C for later use;

[0035] (2) Take 160ml of white oil, 10ml of Span-80 and 7ml of Tween-80, mix well with a stirrer at 200rpm / min for 4min, and bathe in water at 60°C for 20min;

[0036] (3) Take 20ml of EMULSIGEN (EM) and the solution obtained in step (2), stir with a stirrer at 200rpm / min for 4min to mix evenly, and store at 4°C for later use;

[0037] (4) Add 80ml of the CpG ODN solution obtained in step (1) into the solution in step (3), then stir with a stirrer at 600rpm / min for 4min to emulsify and mix evenly, and place at room temperat...

Embodiment 2

[0040] Prepare raw materials according to the following ratios:

[0041]

[0042] The PBS composition is: by weight percentage, 0.7% NaCl, 0.015% KCl, 0.013% Na 2 HPO 4 , 0.0145% KH 2 PO 4 Soluble in water; damp heat autoclave: 117°C, 19min.

[0043](1) After CpG ODN was synthesized by TAKARA Company, it was prepared with sterile PBS solution to 22 μg / ml, and stored at -18°C for future use.

[0044] (2) Take 170ml of white oil, 12ml of Span-80 and 6ml of Tween-80, mix well with a stirrer at 250rpm / min for 5min, and bathe in water at 65°C for 25min.

[0045] (3) Take 25ml of EMULSIGEN (EM) and the solution obtained in step (2), stir with a stirrer at 250rpm / min for 5min to mix evenly, and store at 5°C for later use;

[0046] (4) Add 85ml of the CpG ODN solution obtained in step (1) to the solution in step (3), then stir with a stirrer at 650rpm / min for 5min to emulsify and mix evenly, and place at room temperature for 25min.

[0047] Collect the mixed solution, which i...

Embodiment 3

[0049] Prepare raw materials according to the following ratios:

[0050]

[0051] The PBS composition is: by weight percentage, 0.8% NaCl, 0.02% KCl, 0.014% Na 2 HPO 4 , 0.02% KH 2 PO 4 Soluble in water; damp heat autoclave: 118°C, 20min.

[0052] (1) After CpG ODN was synthesized by TAKARA Company, it was prepared with sterile PBS solution to 25 μg / ml, and stored at -20°C for later use;

[0053] (2) Take 172ml of white oil, 11ml of Span-80 and 8ml of Tween-80, mix well with a stirrer at 300rpm / min for 6min, and bathe in water at 68°C for 26min;

[0054] (3) Take 28ml of EMULSIGEN (EM) and the solution obtained in step (2), stir with a stirrer at 280rpm / min for 6min to mix evenly, and store at 6°C for later use;

[0055] (4) Add 90ml of the CpG ODN solution obtained in step (1) into the solution in step (3), then stir with a stirrer at 680rpm / min for 5.5min to emulsify and mix evenly, and place at room temperature for 24min;

[0056] Collect the mixed solution to obta...

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Abstract

The invention discloses a high-efficiency CpG preparation, a preparation method thereof and application. The CpG preparation consists of components including 80-120ml of PBS (phosphate buffer solution) of CpG ODN (oligodeoxynucleotide) with the concentration of 10-45mug / ml, 160-180ml of white oil, 10-14ml of span-80, 6-10ml of tween-80 and 20-40ml of EMULSIGEN. The preparation is quite favorable for strengthening antigen uptake ability of antigen presenting cells, promotes formation of Th1 type immune response, can reduce the application amount of vaccine, improves the service effect of the vaccine, and is higher in stability and more convenient to be stored.

Description

technical field [0001] The invention belongs to the technical field of veterinary vaccines, and relates to a veterinary vaccine adjuvant and its preparation method and application, in particular to a high-efficiency vaccine adjuvant CpG preparation and its preparation method and application. Background technique [0002] Many domestic veterinary vaccine manufacturers use aluminum adjuvants or oil adjuvants for vaccine adjuvants, but these adjuvants have certain side effects. Oil emulsions stimulate local granulomas or inflammatory reactions: aluminum adjuvants can cause injection site The local inflammatory reaction will cause granuloma, and some individual animals will cause stress response and lead to individual death. And the induced immune response belongs to Th2 type, which is not enough to completely protect animals against infection. Although it can enhance humoral immunity, the antibody is mainly IgGⅠ, which cannot significantly improve cell-mediated immune response...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K39/39A61K47/44
Inventor 张玲华李华周姜增固
Owner SOUTH CHINA AGRI UNIV
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