Method for preparing avian infectiousness bursa of fabricius virus transfer resistant factor
A chicken infectivity and transfer factor technology, which is applied in the preparation methods of peptides, antiviral agents, chemical instruments and methods, etc., can solve the problems of large loss of product activity, incomplete cell fragmentation, and difficulty in improving yield, etc. Reduced morbidity, no adverse reactions, and easy-to-source effects
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Embodiment 1
[0028] Preparation of Anti-Chicken Infectious Bursal Virus Transfer Factor
[0029] 1. Preparation of virus antigen: inoculate chicken infectious bursal virus into 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, and crush it by ultrasonic , 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 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;
[0031] 3. Immune pigs with the above-...
Embodiment 2
[0042] Preparation of Anti-Chicken Infectious Bursal Virus Transfer Factor
[0043] (1) Preparation of raw materials: 10 healthy 6-month-old pigs with no medical history were selected and immunized with chicken infectious bursal virus vaccine subcutaneously. After 15 days, they were immunized again. The method was the same 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;
[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 get homogenate;
[0046] (4) Freezing and thawing: place the homogena...
Embodiment 3
[0053] Detection of the anti-chicken infectious bursal virus transfer factor prepared by the process of the present invention
[0054] The anti-chicken infectious bursal 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 1.460mg / ml as determined by the biuret method.
[0059] (5) Determination of nucleic acid content: The nucleic acid content of this product was determined by the orcinol method to...
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