Preparation method for N-2-hydroxypropyldimethylethyl ammonium chloride chitosan/carboxymethyl chitosan Newcastle disease virus nanoparticles
A technology of hydroxypropyl dimethyl ethyl ammonium chloride and epoxypropyl dimethyl diethyl ammonium chloride, applied in N-2-hydroxypropyl dimethyl ethyl ammonium chloride chitosan /In the field of preparation of carboxymethyl chitosan Newcastle disease virus nanoparticles, it can solve the problems of poor adsorption capacity on mucosal surface, low vaccine antigen encapsulation rate and drug loading capacity, poor solubility of chitosan, etc.
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specific Embodiment approach 1
[0024] Specific embodiment one: the preparation method of present embodiment N-2-hydroxypropyl dimethyl ethyl ammonium chloride chitosan / carboxymethyl chitosan Newcastle disease virus nanoparticle, carry out according to the following steps:
[0025] One, the preparation of N-2-hydroxypropyl dimethyl ethyl ammonium chloride chitosan (N-2-HFCC);
[0026] Two, the preparation of N-2-hydroxypropyl dimethyl ethyl ammonium chloride chitosan / carboxymethyl chitosan Newcastle disease virus nanoparticles (NDV / LaSota-N-2-HFCC / CMC-NPs):
[0027] a. Take 0.5 ~ 2.0mg / mL N-2-hydroxypropyldimethylethylammonium chloride chitosan solution 5mL, drop 2mL of Newcastle disease L-series virus solution into it, stir magnetically for 3min and mix evenly to obtain a solution A;
[0028] b. Stir solution A magnetically at 900-1400 r / min for 10-50 min at room temperature under sterile conditions, then add 0.5-3.0 mg / mL N,O-carboxymethyl chitosan solution 1.25-7.5 mL, to obtain solution B;
[0029] c....
specific Embodiment approach 2
[0032] Specific embodiment two: the present embodiment is different from specific embodiment one: the preparation method of N-2-hydroxypropyl dimethyl ethyl ammonium chloride chitosan in the step one is as follows:
[0033] One, the synthesis of epoxypropyl dimethyl ethyl ammonium chloride (EEA):
[0034]Add epichlorohydrin to a dry three-neck flask equipped with an electric stirrer, dropping funnel and condensing reflux device at 4-15°C, stir continuously and add N,N-dimethylethylamine dropwise, and finish adding within 1 hour, at the same time Heat to 10-35°C, react at constant temperature for 4-14 hours, stop the reaction, wash the product with ice acetone, and filter it with suction, put the filtered product in a watch glass, and dry it in a vacuum dryer for 6-24 hours to obtain ring Oxypropyldimethylethylammonium chloride; wherein the ratio of epichlorohydrin to N,N-dimethylethylamine is 2 to 6:1;
[0035] Two, the preparation of N-2-hydroxypropyl dimethyl ethyl ammonium...
specific Embodiment approach 3
[0044] Specific embodiment three: this embodiment is different from specific embodiment one or two: the virus content of the Newcastle disease L-line virus liquid in step 2a is 10 7.5 EID 50 . Others are the same as in the first or second embodiment.
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