Preparation of anti-newcastle disease virus transfer factor

A technology of chicken Newcastle disease virus and transfer factor, which is applied in the field of preparation of anti-Newcastle disease virus transfer factor, can solve the problems of large loss of product activity, incomplete cell fragmentation, and difficulty in improving the yield, so as to reduce the incidence rate, improve the cure rate, source easy effect

Inactive Publication Date: 2008-11-05
TIANJIN SHENGJI GRP CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The content of the above patent application is to directly filter and extract the broken tissue after freezing and centrifuging, which also has the defect of incomplete cell crushing, and the process is cumbersome, the time is long, and the activity loss of the product is also large. Therefore, the yi

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Example Embodiment

[0027] Example 1:

[0028] Preparation of transfer factor against chicken Newcastle disease virus

[0029] 1. Preparation of virus antigen: inoculate chicken embryo allantoic fluid with chicken Newcastle disease virus at the age of 10 days at 33°C for 48-72 hours, take chicken embryo allantoic fluid, put it in a thick-walled bottle, ultrasonically break it, and centrifuge at 5000 rpm for 20 minutes , Discard the sediment and leave the supernatant;

[0030] 2. Take the centrifugal supernatant, and then discontinuous gradient density centrifugation to purify the virus antigen: After centrifuging the virus antigen, take the supernatant and fill it with discontinuous gradient density prepared with 15%, 30%, 45% and 60% sucrose Centrifuge tube, ultracentrifugation, 165,000 rpm for 3 hours, aspirate according to the position of the centrifuge tube where the chicken Newcastle disease virus is located for use;

[0031] 3. Immunize pigs with the virus antigen extracted above: immunize pigs...

Example Embodiment

[0041] Example 2:

[0042] Preparation of transfer factor against chicken Newcastle disease virus

[0043](1) Preparation of raw materials: 10 healthy 6-month-old pigs with no history of disease were selected and injected subcutaneously with chicken Newcastle disease virus vaccine. 15 days later, immunization was performed again, and the method and dosage were the same as above. 15-20 days after the second immunization, the spleen and thymus were slaughtered, and the spleen and thymus were taken on ice, and transferred and stored at -20°C for later use.

[0044] (2) Pretreatment: After weighing the pig spleen, wash the pig spleen with cold three-distilled water, cut off its fascia and fatty tissue with scissors, and then wash it with cold three-distilled water.

[0045] (3) Fragmentation: Cut the cleaned pig spleen into pieces, add 2 times the volume of cold physiological saline, and mash 3 times with a high-speed tissue masher (1000r / min) under freezing conditions, 3 minutes each...

Example Embodiment

[0052] Example 3:

[0053] Detection of anti-chicken Newcastle disease virus transfer factor prepared by the process of the present invention

[0054] The anti-chicken Newcastle disease virus transfer factor (hereinafter referred to as "this product") prepared by the process described in Example 1 was tested as follows:

[0055] (1) Ultraviolet spectrophotometry: This product has a high absorption peak at 250.0-252.0nm, and ABS260 / ABS280>2.0.

[0056] (2) The standard solution is light yellow, and the pH value is between 6.0-6.5.

[0057] (3) 20% sulfosalicylic acid test: This product has no turbidity and precipitation, indicating that the protein reaction is negative, and it does not contain macromolecular proteins.

[0058] (4) Determination of peptide content: The peptide content of this product is 1.260mg / ml determined by the biuret method.

[0059] (5) Determination of nucleic acid content: The nucleic acid content of this product is 631.56ug / ml determined by the lichenol met...

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Abstract

The invention discloses a preparation method for transferring gene of a natural effective pure biological active matter for resisting a newcastle disease virus. The invention is carried out according to the following steps: inoculating the newcastle disease virus into a chick embryo allantois liquid of ten days to prepare a virus antigen; extracting the virus antigen; using the extracted virus antigen for immunizing a pig; extracting the transferring gene for resisting the newcastle disease virus from the immunized pig; therefore, the transferring gene of the invention can be used for playing the role of preventing the infection of the newcastle disease virus and protecting a normal organism cell from being infected by the newcastle disease virus, thereby reducing the disease rate.

Description

technical field [0001] The invention relates to the field of medicines and preparations thereof, in particular to a preparation method of anti-chicken Newcastle disease virus transfer factor. Background technique [0002] Chicken Newcastle disease is an acute respiratory infectious disease caused by chicken Newcastle disease virus. The biggest feature of NDV is that it is prone to mutation, which causes the anti-NDV specific antibodies that have appeared in chickens to lose their ability to neutralize the virus, thus making chickens susceptible to infection and even causing large-scale epidemics. Sick chickens suffer from symptoms such as diarrhea and dyspnea after being infected by Newcastle disease virus, and severe cases can lead to death. [0003] Although there are some anti-Newcastle disease virus medicines or preparations at present: but because the specificity of the existing medicine preparations is not strong, the curative effect is limited, and the therapeutic pr...

Claims

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Application Information

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IPC IPC(8): C07K1/14C07K1/34A61P31/14
Inventor 田杏芳王连民
Owner TIANJIN SHENGJI GRP CO LTD
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