Preparation method of animal interferon

A technology of interferon and animals, which is applied in the field of animal medicine, can solve the problems of low interferon content and no reports, and achieve the effects of low dosage, low preparation cost and simple production process

Inactive Publication Date: 2012-11-21
TIANJIN ASIAN BIOENG
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] The problem is that in the production of chicken interferon, the conventional use of chicken Newcastle disease virus (NDV) to induce chicken spleen leukocytes to produce low levels of interferon, but no one uses Sendai virus as an interferon inducer
[0005] However, there is no report on the efficient preparation of interferon using Sendai virus and astragalus polysaccharide as compound interferon inducer

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0020] The concrete steps of a kind of preparation method of animal interferon are as follows:

[0021] (1) Chicken spleen lymphocyte preparation

[0022] Aseptically take the spleen of the experimental chicken, prepare the lymphocyte suspension with the spleen, add 10 μg / ml kanamycin, dilute the lymphocyte suspension with the prepared culture medium, and dilute the chicken spleen lymphocyte concentration to 1×106 per ml . The isolation of chicken spleen lymphocytes should be carried out within 48 hours after collection. The number of living cells should reach more than 90%. And the lymphocyte monolayer was subcultured; the chicken spleen lymphocyte concentration was diluted to contain 1×10 per ml 6 . Isolation of chicken spleen lymphocytes should be performed within 48 hours of preparation. The number of living cells should reach more than 90%.

[0023] (2) induced virus

[0024] Sendai virus is used, and the hemagglutination titer can only be put into production when ...

Embodiment 2

[0044] The concrete steps of a kind of preparation method of animal interferon are as follows:

[0045] (1) Aseptically take experimental chicken spleen, prepare lymphocyte suspension with spleen, 10 μg / ml penicillin and streptomycin, dilute cell sediment with prepared 1640 medium, so that chicken spleen lymphocyte concentration is diluted to contain 1×10 7 .

[0046] (2) Add 8% chicken plasma to chicken spleen lymphocyte suspension and mix well.

[0047] (3) A small amount of interferon was added to the above chicken spleen lymphocyte suspension to make the final concentration 200-300 IU / ml, and cultured in a water bath at 37° C. for 1 hour.

[0048] (4) Add astragalus polysaccharides to make the final concentration 25 μg / ml; incubate in a water bath at 37° C. for 2 hours.

[0049] (5) Add 512 hemagglutination units / ml of Sendai virus to the activated chicken spleen lymphocyte suspension for induction, and culture on a shaker at 37° C. for 12-24 hours.

[0050] (6) When t...

Embodiment 3

[0055] A preparation method of animal interferon, the specific steps are as follows:

[0056] (1) Aseptically collect experimental chicken spleen, prepare lymphocyte suspension with spleen, add 10 μg / ml kanamycin, and place in refrigerator at 4.0°C. Dilute the cell pellet with the prepared culture medium, so that the chicken spleen lymphocyte concentration is diluted to 1×10 per ml 5 .

[0057] (2) A small amount of interferon was added to the chicken spleen lymphocyte suspension to make the final concentration 800 IU / ml, and cultured in a water bath at 37°C for 2 hours.

[0058] (3) Astragalus polysaccharide was added to make the final concentration 50 μg / ml, and cultured in a water bath at 37° C. for 2 hours.

[0059] (4) Add 256-1280 hemagglutination units / ml Sendai virus again in the activated chicken spleen lymphocyte suspension to induce, and add the DMEM medium containing 5% bovine serum in the chicken spleen lymphocyte suspension simultaneously, 37 Cultivate on a shak...

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PUM

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Abstract

The invention relates to a preparation method of animal interferon. The preparation method comprises the following implementation steps: (1) acquiring experiment chicken spleen in a sterile way, preparing lymphocyte suspension liquid by the spleen, adding 10mug / ml of kanamycin, and diluting the lymphocyte suspension liquid by the prepared culture solution; (2) adding a small quantity of interferon containing 105 into each ml of the chicken spleen lymphocyte, and cultivating by means of water bath under 37 DEG C for 0.5-2h; (3) adding astragalus polysaccharides so as to leading the final concentration of the astragalus polysaccharides to be 25-100mug / ml, and cultivating by means of water bath under 37 DEG C for 1-2h; (4) adding proper interferon inducer sendai virus into the chicken spleenlymphocyte suspension liquid; and (5) collecting supernate in a centrifuging way when the interferon reaches a peak value so as to obtain a coarse animal interferon. The interferon prepared by the method is high in content and lower in preparation cost obviously.

Description

technical field [0001] The invention belongs to the field of animal medicine, in particular to a method for preparing animal interferon. Background technique [0002] Chicken viral diseases are still the main infectious diseases that seriously restrict the healthy development of the chicken industry. However, traditional infectious disease treatment programs are sometimes difficult to work clinically, especially for viral diseases. So far, there are no effective therapeutic drugs and methods for chicken viral diseases. In recent years, there have been experiments on the prevention and treatment of chicken viral diseases with Chinese herbal medicine preparations, but the cure rate is low. Antibiotics and sulfa drugs are basically ineffective. At present, chicken interferon is an effective drug for the treatment of chicken viral diseases. [0003] The existing problem is: In the production of chicken interferon, the newcastle disease virus (NDV) is used routinely to induce...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K38/21C12P21/02
Inventor 王文彪王丽莉
Owner TIANJIN ASIAN BIOENG
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