Method for preparing avian infectious brunchitis virus transfer resistant factor

A bronchitis and chicken infectious technology, applied in the field of preparation of anti-chicken infectious bronchitis virus transfer factor, can solve the problems of incomplete cell fragmentation, large loss of product activity, difficulty in improving yield, etc., and achieve no adverse reactions , reduce the incidence of disease, the effect of easy source

Inactive Publication Date: 2008-10-29
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
[0008] 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

[0030] Preparation of Anti-Chicken Infectious Bronchitis Virus Transfer Factor

[0031] 1. Preparation of virus antigen: Inoculate chicken infectious bronchitis virus in the allantoic fluid of 10-day-old chicken embryo, 48-72 hours at 33°C, take the allantoic fluid of chicken embryo, put it in a small thick-walled bottle, and smash it by ultrasonic. Centrifuge at 5000rpm for 20 minutes, discard the sediment and keep the supernatant;

[0032] 2. Take the centrifuged supernatant, then discontinuous gradient density centrifugation and purification to extract the viral antigen; after the viral antigen is centrifuged, take the supernatant and fill it with discontinuous gradient density prepared with 15%, 30%, 45% and 60% sucrose Centrifuge tube, ultracentrifuge, 165,000 rpm for 3 hours, determine the position of the centrifuge tube where the virus is located according to the molecular weight of different viruses, and suck it out for later use;

[0033] 3. Immune pigs with the abov...

Embodiment 2

[0044] Preparation of Anti-Chicken Infectious Bronchitis Virus Transfer Factor

[0045] (1) Preparation of raw materials: 10 healthy 6-month-old pigs with no medical history were selected and immunized with chicken infectious bronchitis virus vaccine subcutaneously. After 15 days, they were immunized again with the same method as above. Slaughter 15-20 days after the second immunization, take the spleen and thymus on ice, transfer and store at -20°C for later use;

[0046] (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;

[0047] (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 get homogenate;

[0048] (4) Freezing and thawing: place the homo...

Embodiment 3

[0055] Detection of anti-chicken infectious bronchitis virus transfer factor prepared by process of the present invention

[0056] The anti-chicken infectious bronchitis virus transfer factor (hereinafter referred to as "this product") prepared by the process described in embodiment 1 is detected as follows:

[0057] (1) Ultraviolet spectrophotometric measurement: This product has a high absorption peak at 250.0-252.0nm, and ABS260 / ABS280>2.0.

[0058] (2) The standard solution is light yellow and the pH value is between 6.0-6.5.

[0059] (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.

[0060] (4) Determination of peptide content: The peptide content of this product is 1.760mg / ml as determined by the biuret method.

[0061] (5) Determination of nucleic acid content: The nucleic acid content of this product was determined to be 657.56ug / ml by ...

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PUM

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Abstract

The invention discloses a preparation method of a natural high-effective biological active substance anti-avian infectious bronchitis virus (IBV) transfer factor. The preparation method comprises the following steps of: inoculating IBV into an allantoic fluid obtained from a 10-day-aged chick embryo, and preparing IBV antigen; extracting the IBV antigen; immunizing a pig with the extracted IBV antigen; and extracting the anti-IBV transfer factor from the immunized pig. The inventive anti-IBV transfer factor can prevent IBV infection and protect normal body cells from being infected by the IBV, so as to reduce incidence 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 infectious bronchitis virus transfer factor. Background technique [0002] Chicken infectious bronchitis is an acute, highly contagious infectious disease caused by infectious bronchitis virus. The disease occurs frequently in winter and spring, and the main clinical manifestation is respiratory disorder. Young chickens are the most susceptible and have a high mortality rate; infection of adult chickens can significantly reduce the egg production rate and cause major economic losses. Comprehensive measures must be taken to prevent and control the production. [0003] The disease can occur in chickens of various ages, but it is the most serious in young chickens, with a higher incidence rate and a mortality rate of 20.0% to 25.0%. Among them, the respiratory tract lesion type is the most serious in chickens under 6 weeks old,...

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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