Biological product for treating gosling plague and preparing method thereof
A biological product and goose plague technology, applied in biochemical equipment and methods, medical preparations containing active ingredients, microorganisms, etc., can solve the problem of low curative effect of goose plague treatment drugs
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Embodiment 1
[0026] Embodiment 1 (preparation of gosling plague egg yolk antibody IgY)
[0027] (1) Preparation of highly immune eggs: immunize laying hens with gosling plague inactivated vaccine, the immunization dose is 1ml / hen, and the immunization interval is 14 days. Eggs can be collected with a titer higher than 1:64.
[0028] (2) Extraction and preparation of egg yolk antibody IgY: immerse high-immune eggs in 1% bromogeramine solution, soak and sterilize for 15 minutes, and dry for later use; mechanically beat the eggs, separate and collect the egg yolks; emulsify with a colloid mill, add vinegar at a ratio of 1:4 Salt buffer solution (pH5.2), stirred for 60 minutes, filtered with a plate and frame filter; added 2% octanoic acid to the filtered supernatant, stirred vigorously for 60 minutes, and filtered with a plate and frame; the filtered supernatant was sterilized with a 0.22 μm microporous filter .
Embodiment 2
[0029] Embodiment 2 (preparation of gosling plague VP3 protein antigen)
[0030] Using the gene cloning technology, the gosling plague VP3 gene was cloned into the prokaryotic expression vector, transfected into the prokaryotic expression host, and induced to express. Afterwards, the VP3 protein was purified by affinity chromatography. The specific operation method is: use the VP3 co-conserved fragment of the classic strain of goose plague virus in GenBank, use the software Premier 5.0 to design and synthesize the upstream and downstream primers of the VP3 gene; take the liver tissue of goose infected with goose plague virus, grind and homogenate Afterwards, total RNA was extracted by conventional RNA extraction technology and reverse-transcribed into cDNA; using cDNA as a template, the VP3 gene was amplified and recovered by using the upstream and downstream primers of the VP3 gene; the recovered product and the expression vector used used the same restriction endogenous Dic...
Embodiment 3
[0031] Embodiment 3 (titer determination of gosling plague VP3 protein antigen and gosling plague egg yolk antibody)
[0032]Use the agar diffusion test team to determine the titer of gosling plague VP3 protein antigen and gosling plague egg yolk antibody. The specific operation is as follows: prepare an agar plate with 8% NaCl and 1% agarose; use plum blossom holes to punch holes on the agar plate;
[0033] (1) Detection of VP3 protein antigen titer: After doubling dilution of VP3 protein antigen, inject it into the peripheral wells of the plum blossom holes, and add gosling plague standard positive serum in the central well, and the sample volume should be filled without overflowing. Put the agar plate into a wet box and act at 37°C for 24h to 48h. The maximum dilution factor at which precipitation lines appear in the central well and the peripheral well is the antigen titer of the VP3 protein.
[0034] (2) Detection of yolk antibody titer: after doubling dilution of yolk a...
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