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Preparation method of PLGA swine influenza DNA vaccine microsphere

A technology of DNA vaccine and swine flu, which is applied in the field of preparation of DNA vaccine microspheres, can solve the problems of short duration and easy degradation of swine flu DNA vaccine, prolong the time of drug action, and the preparation method is simple, easy, and highly efficient. The effect of encapsulation rate

Inactive Publication Date: 2012-01-18
HEILONGJIANG UNIV
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Problems solved by technology

[0004] The purpose of the present invention is to solve the problem that the current swine influenza DNA vaccine has a short duration, is easily degraded in the body, and cannot be sustained release, and provides the preparation of PLGA (polylactic acid / glycolic acid copolymer) swine influenza DNA vaccine microspheres method

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  • Preparation method of PLGA swine influenza DNA vaccine microsphere
  • Preparation method of PLGA swine influenza DNA vaccine microsphere
  • Preparation method of PLGA swine influenza DNA vaccine microsphere

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specific Embodiment approach 1

[0018] Specific embodiment one: the preparation method of the PLGA porcine influenza DNA vaccine microsphere of the present embodiment is prepared according to the following steps:

[0019] 1. Weigh 25mg of PLGA and dissolve it in 0.75mL of dichloromethane. After it is completely dissolved, add 40μl of swine influenza virus H3N2 subtype HA gene recombinant plasmid DNA solution, and ultrasonic emulsify for 10s under ice bath conditions to obtain colostrum. Add 20 mL of polyvinyl alcohol with a mass concentration of 2% at a rotating speed of 7000 r / min to obtain double emulsion;

[0020] 2. Magnetically stir the double emulsion for 5 hours at a speed of 200r / min, then centrifuge at a speed of 4000r / min for 10 minutes, collect the microspheres and wash them with sterilized water for 3 times, and then place them at -10~-50℃ Vacuum freeze-drying under the condition of PLGA porcine influenza DNA vaccine microspheres is completed; wherein step 1 has 2 mg of porcine influenza virus H3...

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Abstract

A preparation method of PLGA swine influenza DNA vaccine microspheres relates to a preparation method of DNA vaccine microspheres. The invention solves the problem that present swine influenza DNA vaccines have a short duration, are easily degraded in vivo, and can not release sustainedly. The method comprises the following steps: firstly, dissolving PLGA in dichloromethane, adding a swine influenza H3N2 subtype HA gene recombinant plasmid DNA solution, adding polyvinyl alcohol after emulsification to obtain a multiple emulsion; secondly, performing magnetic stirring and centrifugation of the multiple emulsion, collecting microspheres and washing, performing vacuum freeze drying. The PLGA swine influenza DNA vaccine microsphere prepared by the invention has a uniform size, a round profile, a smooth surface, a particle size of 6.08+ / -2.34 microns, average entrapment efficiency of 60%+ / -3.8%, and an average drug loading of 5.5%+ / -0.5%; with the guarantee of the stability of the DNA vaccine, the microsphere has high entrapment efficiency, can maintain an optimal drug concentration in vivo, prolong the drug action time, and realize the sustained release of the drug.

Description

technical field [0001] The invention relates to a preparation method of DNA vaccine microspheres. Background technique [0002] Swine influenza is an acute and contagious respiratory disease of pigs caused by influenza A virus of the family Orthomyxoviridae. It spreads rapidly and often causes outbreaks of acute respiratory diseases. Because there are many subtypes of swine influenza virus, antigens are easy to mutate and can be transmitted between species, it brings many challenges to the prevention of influenza diseases and even human health. Therefore, swine flu plays a key role between human flu and bird flu, and has great public health significance. [0003] DNA vaccine is to directly inoculate the body with the eukaryotic expression plasmid containing the gene encoding the antigen, and express the corresponding antigen in the body to stimulate the body to generate an immune response against the antigen and produce protective immunity. At present, the results of large...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K48/00A61K39/145A61K47/34A61K9/14A61P31/16A61J3/00
Inventor 赵凯
Owner HEILONGJIANG UNIV
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