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Preparation method of N-2-hydroxypropyl trimethyl ammonium chloride chitosan/N, O-carboxymethyl chitosan newcastle disease attenuated live vaccine nanoparticle

A technology of hydroxypropyltrimethylammonium chloride and carboxymethyl chitosan, which is applied in antiviral agents, viral antigen components, freeze-dried transportation, etc., can solve the problems of chitosan application limitations and achieve good biological Adhesiveness, uniform particle size distribution, and low drug side effects

Active Publication Date: 2013-03-13
HEILONGJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0005] As the only natural polysaccharide with cations in nature, chitosan has special functions such as good biological activity, biocompatibility, degradability and antibacterial properties. However, chitosan can only be dissolved in acidic solution. Applications in this field are severely limited

Method used

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  • Preparation method of N-2-hydroxypropyl trimethyl ammonium chloride chitosan/N, O-carboxymethyl chitosan newcastle disease attenuated live vaccine nanoparticle
  • Preparation method of N-2-hydroxypropyl trimethyl ammonium chloride chitosan/N, O-carboxymethyl chitosan newcastle disease attenuated live vaccine nanoparticle
  • Preparation method of N-2-hydroxypropyl trimethyl ammonium chloride chitosan/N, O-carboxymethyl chitosan newcastle disease attenuated live vaccine nanoparticle

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specific Embodiment approach 1

[0017] Specific embodiment one: present embodiment N-2-hydroxypropyltrimethylammonium chloride chitosan / N, the preparation method of O-carboxymethyl chitosan Newcastle disease live attenuated vaccine nanoparticle, according to the following steps accomplish:

[0018] 1. Take 5mL of N-2-hydroxypropyltrimethylammonium chloride chitosan solution with a concentration of 0.5-20mg / mL, and then add 1.25-7.5mL of Newcastle disease L-series virus solution dropwise to it, and stir magnetically for 3min , to obtain solution A;

[0019] 2. Solution A is magnetically stirred at 900-1300r / min for 10-40min at room temperature and under sterile conditions, and then 2mL of N,O-carboxymethyl chitosan solution with a concentration of 0.5-3.0mg / mL is added dropwise to obtain a solution B;

[0020] 3. Centrifuge solution B at 4°C and 12000r / min for 2min, take the precipitate and wash it three times with deionized water, then vacuum freeze-dry for 24h to obtain N-2-hydroxypropyltrimethylammonium...

specific Embodiment approach 2

[0028]Specific embodiment two: the difference between this embodiment and specific embodiment one is that getting 5mL concentration in step one is the N-2-hydroxypropyltrimethylammonium chloride chitosan solution of 1mg / mL, then to wherein dropwise 2mL Newcastle disease L-series virus solution. Other steps and parameters are the same as those in Embodiment 1.

specific Embodiment approach 3

[0029] Specific embodiment 3: The difference between this embodiment and specific embodiment 1 is that in step 2, solution A is magnetically stirred at 1200 r / min for 30 min at room temperature and under sterile conditions, and then 2 mL of N with a concentration of 1 mg / mL is added dropwise. carboxymethyl chitosan solution. Other steps and parameters are the same as those in Embodiment 1.

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Abstract

The invention relates to a preparation method of a newcastle disease attenuated live vaccine nanoparticle, in particular to a preparation method of a N-2-hydroxypropyl trimethyl ammonium chloride chitosan / N, O-carboxymethyl chitosan newcastle disease attenuated live vaccine nanoparticle, which comprises the following steps of: 1) dripping newcastle disease L-series virus fluid in N-2-hydroxypropyl trimethyl ammonium chloride chitosan solution to obtain solution A; 2) mixing the solution A, dripping N, O-carboxymethyl chitosan solution to obtain solution B; and 3) centrifuging the solution B, sedimenting, washing by deionized water, and carrying vacuum freezing and drying to obtain the product. According to the N-2-hydroxypropyl trimethyl ammonium chloride chitosan / N, O-carboxymethyl chitosan newcastle disease attenuated live vaccine nanoparticle prepared by the invention, the particle size is easy to control, the particle size distribution is uniform, the maximum encapsulation rate can reach 98%, the medicine carrying quantity is large, and the compatibility and the biology adherence property are good. The preparation method is simple and easy to implement, the condition is mild, the toxic side effects of the medicine are fewer, the sustained-release time is long, the production cost is lower, and the large scale production is easy to implement.

Description

technical field [0001] The invention relates to a preparation method of nano particles of live attenuated Newcastle disease vaccine. Background technique [0002] Newcastle disease (ND) is an acute, highly contagious, fatal poultry infectious disease caused by Newcastle disease virus (NDV). It can cause 100% morbidity and mortality in other birds such as parrots. The main features of the disease are dyspnea, diarrhea, nervous disorder, mucosal and serosal hemorrhage, acute onset and high fatality rate. It is one of the most serious infectious diseases that endanger the poultry industry. Currently, the most important and effective means of prevention and control of the disease is vaccination. However, the common disadvantage of existing vaccines is that their immunogenicity is weak, and strong immune adjuvants or appropriate mucosal immune delivery systems are needed to effectively induce immune responses. Nano-vaccine is a new slow-release vaccine that uses degradable bio...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K9/19A61K39/17A61P31/14
Inventor 赵凯金政张旭陈刚吴昊
Owner HEILONGJIANG UNIV
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