Newcastle disease vaccine Yupingfeng polysaccharide immunopotentiator
An immune enhancer, jade screen technology, applied in the field of livestock and poultry immunity, can solve the problems of lack of immune enhancer, short duration of efficacy, long medication time, etc., to enhance animal immune function, promote value, and improve disease resistance. Effect
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Embodiment 1
[0029] The preparation of embodiment 1 polysaccharide
[0030] Based on 1000g of Yupingfeng Powder, astragalus, Atractylodes macrocephala, and Fangfeng are configured according to the mass ratio of 3:1:1 to make 1000g of Yupingfeng Powder, add 8 times the quality of water and soak for 30 minutes; then boil for 1 hour, add 6 times the quality of the medicinal residue after filtering Boil water for 1 hour, add 6 times the quality of water to the filter residue, boil for 1 hour and filter, combine the filtrate three times, heat and concentrate the filtrate to 1000mL, after cooling, centrifuge at 3000rpm for 10min, remove impurities, distill water to make 1000mL; use one-step alcohol precipitation method Extraction, slowly add 95wt% ethanol to the above-mentioned concentrated solution to make the final concentration of ethanol in the mixed solution reach 80wt%, stir while adding, let stand for 24 hours after the addition is completed, centrifuge, take the precipitate and place it i...
Embodiment 2
[0031] The purification of embodiment 2 polysaccharides
[0032] Take YPF-PS respectively tc Dissolve 500mg in 10mL of distilled water to obtain 50mg.mL -1 Sample, followed by chromatography column (1.6cm×90cm), the medium is SephadexA-25, the flow rate is 0.42mL min -1 Load the sample, and then collect it in fractions, using 0.05mol·L -1 NaCl and 1mol L -1 Gradient elution with 400mL of NaCl each, and the eluate was dialyzed through a dialysis bag to remove sodium chloride and impurities to obtain purified polysaccharide (YPF-PS tp ).
Embodiment 3
[0033] The comparison of embodiment 3 in vitro immune effects
[0034] Method: Yupingfeng polysaccharide was used as test material, and RAMPS tc , RAMPS tp , Atractylodes macrocephala graded polysaccharides (RAMPS 60c , RAMPS 70c and RAMPS 80c) as comparison. Firstly, the safe concentrations of the three prescriptions on peripheral blood lymphocytes were determined, and then the three prescriptions were diluted with cell culture medium (RPMI1640) to 62.5, 31.25, 15.625, 7.813, and 3.907 μg·mL -1 A total of 5 working concentrations were added to the cultured chicken peripheral blood lymphocytes respectively, and the absorbance value at 570nm was detected on an enzyme-linked immunosorbent analyzer (A 570value), as an indicator of lymphocyte proliferation. And calculate the highest cell proliferation rate of each polysaccharide according to the formula: the highest cell proliferation rate=(the highest A of the polysaccharide group 570 Value - Cell Control A 570 value) / cel...
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