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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 yield hard to improve
[0006] Porcine spleen and thymus are places where immunocompetent cells gather, and the sources of materials are very extensive. Therefore, it is of great theoretical and practical significance to extract specific transfer factors from these cells, but it has not been reported yet.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] Preparation of Anti-Newcastle Disease Virus Transfer Factor

[0029] 1. Preparation of virus antigen: Inoculate chicken Newcastle disease virus in the allantoic fluid of 10-day-old chicken embryos, 48-72 hours at 33°C, take the allantoic fluid of chicken embryos, put it in a small thick-walled bottle, smash it by ultrasonic, and centrifuge at 5000rpm for 20 minutes , discard the sediment and leave the supernatant;

[0030] 2. Take the centrifuged supernatant, then discontinuous gradient density centrifugation and purification to extract the virus antigen: after centrifuging the viral antigen, take the supernatant and put it into the discontinuous gradient density prepared with 15%, 30%, 45% and 60% sucrose Centrifuge tube, ultracentrifuge, 165,000 rpm for 3 hours, suck out according to the position of the centrifuge tube where the Newcastle disease virus is located;

[0031] 3. Immune pigs with the above-mentioned extracted virus antigens: use the above-mentioned extra...

Embodiment 2

[0042] Preparation of Anti-Newcastle Disease Virus Transfer Factor

[0043](1) Preparation of raw materials: 10 healthy 6-month-old pigs with no medical history were selected and immunized with Newcastle disease virus vaccine subcutaneously. After 15 days, they were immunized again with the same method as above. They were slaughtered 15-20 days after the second immunization, and the spleen and thymus were taken out, stored on ice, 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 adipose tissue with scissors, and then wash it with cold three-distilled water.

[0045] (3) Crushing: Cut the pig spleen washed above into pieces, add 2 times the volume of cold normal saline, and mash it with a high-speed tissue masher (1000r / min) for 3 times under freezing conditions, 3 minutes each time, to prepare Obtain a homogenate.

[0046] (4) Freezing and thawing: put the ...

Embodiment 3

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

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

[0055] (1) Ultraviolet spectrophotometric measurement: 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 1.260 mg / ml as determined by the biuret method.

[0059] (5) Determination of nucleic acid content: The nucleic acid content of this product was determined to be 631.56ug / ml by the orcinol method...

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PUM

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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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