Preparation method of chicken infectious bronchitis N-2-hydroxypropyl trimethyl ammonium chloride chitosan/carboxymethyl chitosan nano living vaccine

A technology of hydroxypropyl trimethyl ammonium chloride and carboxymethyl chitosan, which is applied in the field of preparation of chicken infectious bronchitis nanometer live vaccines, can solve the problem of poor solubility, weak solubility, and chitosan dissolution effect. It can achieve the effect of long release time, reducing the dosage, and improving the immunity of the body.

Inactive Publication Date: 2013-07-24
HEILONGJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Chitosan is widely used because of its non-toxicity, good biocompatibility and biodegradability, but due to its weak solubility limitation (only soluble in solutions with pH less than 6.5, and in alkaline and neutral However, the solubility of chitosan is extremely poor under neutral conditions, that is to say, at physiological pH, the dissolution effect of chitosan is very poor), so its application as a nanoparticle delivery system has certain limitations.

Method used

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  • Preparation method of chicken infectious bronchitis N-2-hydroxypropyl trimethyl ammonium chloride chitosan/carboxymethyl chitosan nano living vaccine
  • Preparation method of chicken infectious bronchitis N-2-hydroxypropyl trimethyl ammonium chloride chitosan/carboxymethyl chitosan nano living vaccine
  • Preparation method of chicken infectious bronchitis N-2-hydroxypropyl trimethyl ammonium chloride chitosan/carboxymethyl chitosan nano living vaccine

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specific Embodiment approach 1

[0019] Specific embodiment one: the preparation method of chicken infectious bronchitis N-2-hydroxypropyltrimethylammonium chloride chitosan / carboxymethyl chitosan nano live seedling of the present embodiment is realized according to the following experimental steps:

[0020] 1. Take 50-400 μl IBV H120 virus liquid and add sterile deionized water to make up to 2mL, and then add 5mL of N-2-hydroxypropyltrimethyl chloride after filtration sterilization with a concentration of 0.5-2.0mg / mL Ammonium chloride chitosan solution, then stirred for 5min under the condition of 300r / min with stirring speed under aseptic conditions at room temperature, to obtain mixed solution A;

[0021] 2. Stir the mixed solution A at room temperature and under sterile conditions at a speed of 900r / min to 1300r / min for 1min, then add 2mL of N, O-carboxymethyl base chitosan solution; then magnetically stirred for 20min to 50min under constant stirring speed to obtain mixed solution B;

[0022] 3. Centri...

specific Embodiment approach 2

[0028] Specific embodiment 2: The difference between this embodiment and specific embodiment 1 is that the method of filter sterilization described in step 1 and step 2 is: filter through a 0.22 μm bacterial filter in a sterile operating bench. Other steps and parameters are the same as those in the first embodiment.

specific Embodiment approach 3

[0029] Specific embodiment 3: The difference between this embodiment and specific embodiment 1 or 2 is that in step 1, 200 μl of virus liquid is taken and 2 mL of sterile deionized water is added to make up 2 mL, and then added to 5 mL of the filtered solution with a concentration of 1.0 mg / mL. N-2-hydroxypropyltrimethylammonium chloride chitosan solution after sterilization. Other steps and parameters are the same as those in Embodiment 1 or 2.

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Abstract

The invention provides a preparation method of a chicken infectious bronchitis N-2-hydroxypropyl trimethyl ammonium chloride chitosan / carboxymethyl chitosan nano living vaccine and relates to a preparation method of a chicken infectious bronchitis nano living vaccine. The invention aims at providing the preparation method of chicken infectious bronchitis N-2-hydroxypropyl trimethyl ammonium chloride chitosan / carboxymethyl chitosan nano living vaccine. The preparation method comprises the following steps of: firstly, preparing a virus solution and adding the virus solution into an N-2-hydroxypropyl trimethyl ammonium chloride chitosan solution after filtration sterilization, and stirring to obtain a mixed solution A; secondly, dropwise adding a carboxymethyl chitosan solution into the mixed solution A after filtration sterilization to obtain a mixed solution B; thirdly, centrifuging the mixed solution B, then washing sediments with sterile deionized water at the temperature of 4 DEG C for three times, and collecting solid-phase materials; and fourthly, adding the solid-phase materials into the sterile deionized water at the temperature of 4 DEG C, then adding 5% of cane sugar skimmed milk solution, and then carrying out vacuum freeze drying, thus the preparation method is completed. The preparation method provided by the invention is applied to the field of veterinary drugs.

Description

technical field [0001] The invention relates to a preparation method of live nano-vaccine of chicken infectious bronchitis. Background technique [0002] Infectious bronchitis is an acute, highly contagious viral disease of chickens caused by infectious bronchitis virus (IBV), characterized by respiratory symptoms, decreased egg production rate, and kidney lesions. Since 1988, it has been popular in most parts of my country, with a morbidity rate of 100% and a mortality rate of 10% to 30%. Due to the pollution of the environment by intensive farming and the continuous emergence of new virus variant strains, and often the genotype performance is different and the serotype variation alternates or occurs simultaneously, and the tissue tropism is also constantly changing, making the immune prevention and control of IBV difficult. , The phenomenon of immune failure occurs from time to time. These two diseases seriously endanger the development of the poultry industry. Timely i...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K39/215A61K47/36A61P31/14
Inventor 赵凯
Owner HEILONGJIANG UNIV
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