Saponin with immunoadjuvant function, preparation method, vaccine preparation containing the saponin as adjuvant and uses thereof
A technology of immune adjuvant and effect, which is applied in the field of saponin with immune adjuvant effect and its preparation and vaccine preparation containing the saponin as adjuvant and application field, which can solve complex chemical structure, variability of aluminum glue, and strong hemolysis and other problems, to achieve the effect of simple preparation process, easy quality and convenient use
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Embodiment 1
[0036] Embodiment 1: preparation of platycodon saponin
[0037] Preparation of Platycodon grandiflora total saponins
[0038] Medicinal platycodon grandiflora coarse powder 5kg, reflux extraction with 70% ethanol for 3 times, 2 hours each time, filter, combine the filtrate, recover ethanol under reduced pressure, add appropriate amount of water to suspend the extract, extract with ether or petroleum ether until the ether liquid is nearly free color. The aqueous layer was extracted with water-saturated butanol or pentanol until the extract was nearly colorless, and the extracts were combined, recovered under reduced pressure and concentrated to obtain the alcohol extract. The alcohol extract is passed through a macroporous adsorption resin column, and the mixture of water and ethanol is used for gradient elution, and the eluent is checked by thin-layer chromatography. The fractions with the same spots are combined, concentrated, and separated to obtain the total saponins of Pl...
Embodiment 2
[0046] Embodiment 2 contains the hepatitis B vaccine preparation of saponin adjuvant
[0047] Weigh 500 μg of platycodon total saponins, platycodon saponin D or platycodon saponin D2, and 100 μg of hepatitis B virus surface antigen (HBsAg), dissolve in 10 ml of normal saline, filter through a 0.22 μm microporous membrane, and aseptically dispense, 1 ml each. Each ml contains saponin 50μg and HBsAg 10μg.
Embodiment 3
[0048] Example 3 Hepatitis B vaccine preparation containing aluminum gel and saponin adjuvant
[0049] Weigh an appropriate amount of platycodon total saponins, platycodon saponin D or platycodon saponin D2, dissolve in physiological saline to make a solution with a concentration of 100 μg / ml, and filter through a 0.22 μm microporous membrane. The saponin solution is mixed with an equal volume of aluminum-containing adjuvanted hepatitis B surface antigen vaccine stock solution (each ml contains 20 μg of HBsAg and 500 μg of aluminum hydroxide), mixed evenly, and aseptically dispensed, 1 ml per tube. Each ml contains saponin 50μg, HBsAg 10μg and aluminum hydroxide 250μg.
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