Preparation of transfer factor against swine fever
A technology of transfer factor and swine fever disease, which is applied in the field of preparation of anti-swine fever transfer factor, can solve the problems of inability to prevent, weak drug competition, and limited curative effect, and achieve long maintenance time and shorten repeated freezing and thawing Time, curative effect and exact effect
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Embodiment 1
[0036] Embodiment 1: Preparation of anti-swine fever transfer factor
[0037] 1. Inoculate CSFV39 (Genbank accession No. J04358) and / or BVDV (Genbank accession No. U63479) in the allantoic fluid of 9-11-day-old chicken embryos and culture at 37°C for 48 hours. Take chicken embryo allantoic fluid, freeze and thaw repeatedly 5 times at -20°C, centrifuge at 5000rpm for 30min, discard the precipitate and collect the supernatant;
[0038] 2. Take the supernatant, and use the sucrose density gradient centrifugation to purify the supernatant to extract the virus. The sucrose concentration is 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, and centrifuged at 150000rpm for 4 hours , according to the molecular weight of the classical swine fever virus, determine the position of the centrifuge tube where the classical swine fever virus is located, and suck it out for later use;
[0039]3. The obtained swine fever virus antigen immunized pigs were selected from healthy immunized pigs, and the ab...
Embodiment 2
[0049] Embodiment 2: Preparation of anti-swine fever transfer factor
[0050] 1. Inoculate classical swine fever virus in the allantoic fluid of 9-11-day-old chicken embryos and culture at 37°C for 72 hours. Take chicken embryo allantoic fluid, freeze and thaw repeatedly 5 times at -20°C, centrifuge at 5000rpm for 30min, discard the precipitate and collect the supernatant;
[0051] 2. Get the supernatant, and use the sucrose density gradient centrifugation to purify the supernatant to extract the virus. The sucrose concentration is 15%, 30%, 45%, 60%, 75%, centrifuged at 180000rpm for 3 hours, and determine the swine fever according to the molecular weight of the swine fever virus The position of the centrifuge tube where the virus is located, aspirate for later use;
[0052] 3. The obtained swine fever virus antigen immunized pigs were selected from healthy immunized pigs, and the above-mentioned extracted virus antigens were subcutaneously injected into the immunized pigs a...
Embodiment 3
[0061] (8) Aliquot and store at 4°C to obtain the stock solution of anti-swine fever virus transfer factor. Embodiment 3: the detection of the anti-swine fever transfer factor prepared by the process of the present invention
[0062] The anti-swine fever transfer factor prepared by the process described in embodiment 1 and 2 is detected as follows:
[0063] 1. Ultraviolet spectrophotometric measurement: the sample obtained above has an absorption peak at 252±2nm, and ABS 260 / ABS 280 >2.0.
[0064] 2. The standard solution is light yellow and the pH value is between 6.0 and 8.0.
[0065] 3. Bacteriological examination: there is no need for aerobic, anaerobic, saprophytic bacteria and fungi in the samples obtained above.
[0066] 4. Detection of 20% sulfosalicylic acid: the sample obtained above has no turbidity and precipitation, indicating that the protein reaction is negative, and it does not contain macromolecular protein.
[0067] 5. Determination of nucleic acid cont...
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