Animal viral vaccine pulsatile release system, and preparation method and application thereof
A technology of pulse release and release system, which can be used in antiviral agents, viral antigen components, pharmaceutical formulations, etc., and can solve the problem of low immunogenicity
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Embodiment 1
[0013] Preparation of Newcastle Disease Inactivated Vaccine Antigen
[0014] Take Newcastle disease attenuated La Sota strain, inoculate susceptible chicken embryos according to conventional methods, after culture, harvest chicken embryo liquid, and inactivate with formaldehyde solution. Take the inactivated Newcastle disease virus chicken embryo liquid, centrifuge at 4°C under sterile conditions to get the supernatant, and then ultrafilter the supernatant through an ultrafiltration membrane with a molecular weight cut-off of 30,000 under sterile conditions at 4°C concentrate. The supernatant after ultrafiltration purification and concentration is the antigen solution for preparing vaccine microspheres.
Embodiment 2
[0016] Preparation I of Newcastle Disease Inactivated Vaccine Microspheres
[0017] Preparation of microspheres: PLGA microspheres of Newcastle disease inactivated vaccine were prepared by drying in double emulsion. The Newcastle disease antigen solution was used as the water phase, and PLGA (50:50) was dissolved in dichloromethane to prepare a solution with an appropriate concentration as the oil phase. Mix the water phase and the oil phase at a ratio of 1:10 (V / V), and ultrasonically emulsify for 10 seconds in an ice bath to obtain colostrum. Add the colostrum into the 2% PVA solution, and stir it through an emulsifier at high speed for 30 seconds at 10,000 rpm to obtain double milk. Transfer the obtained double emulsion to a beaker, stir magnetically at 400rpm for 5 hours, evaporate dichloromethane at room temperature, and then centrifuge at 6000rpm for 10 minutes to collect the microspheres, wash with sterilized water 3 times, and vacuum freeze Dry to get Newcastle disea...
Embodiment 3
[0023] Preparation of Newcastle Disease Inactivated Vaccine Microspheres II
[0024] Preparation of microspheres: PLGA microspheres of Newcastle disease inactivated vaccine were prepared by drying in double emulsion. The Newcastle disease antigen solution was used as the water phase, and PLGA (75:25) was dissolved in dichloromethane to prepare a solution with an appropriate concentration as the oil phase. Mix the water phase and the oil phase at a ratio of 1:10 (V / V), and ultrasonically emulsify for 10 seconds in an ice bath to obtain colostrum. Add the colostrum into the 2% PVA solution, and stir it through an emulsifier at a high speed for 30 seconds at 4000 rpm to obtain double milk. Transfer the obtained double emulsion to a beaker, stir magnetically at 200rpm for 5 hours, evaporate dichloromethane at room temperature, and then centrifuge at 5000rpm for 10 minutes to collect microspheres, wash with sterilized water 3 times, and vacuum freeze Dry to obtain Newcastle disea...
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