Oral nano-vaccine of large yellow croaker and immune type large yellow croaker feed prepared by the same

A nano-vaccine, large yellow croaker technology, applied in vaccines, veterinary vaccines, pharmaceutical formulations, etc., can solve the problems of inability to achieve immune effects, affect immune protection effects, etc., achieve cost-effective, reduce toxicity and stimulation, and biocompatibility Good results

Active Publication Date: 2020-03-31
FUJIAN NORMAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] At present, the research and development of vaccines has developed from traditional all-pathogenic vaccines to using only the required proteins and peptides. Traditional oral vaccines are easily degraded in the complex aquaculture water environment and the enzyme-rich stomach environment, and cannot be delivered well to the rear. The intestine is absorbed by immune cells, which greatly affects the immune protection effect
Cannot achieve ideal immune effect when used alone

Method used

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  • Oral nano-vaccine of large yellow croaker and immune type large yellow croaker feed prepared by the same
  • Oral nano-vaccine of large yellow croaker and immune type large yellow croaker feed prepared by the same
  • Oral nano-vaccine of large yellow croaker and immune type large yellow croaker feed prepared by the same

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preparation example Construction

[0026] The preparation method of described large yellow croaker oral nano-vaccine, it comprises the following processing steps:

[0027] (1) preparing MSN nanoparticles;

[0028] The specific method of step (1) is: add hexadecyltrimethyl-p-toluenesulfonate amine, triethanolamine and water into a round bottom flask, stir and dissolve at 80-85°C for 1-2h; then quickly add tetrasilicic acid Ethyl ester, continue stirring for 2-3h; centrifuge at 12745g for 20-25min to recover the precipitate; use ddH for precipitation 2 Wash twice with O, followed by 6 g / L NH at 60-65 °C 4 NO 3 The methanol solution extracted the product 3 times; after the extraction, the precipitate was recovered by centrifugation at 12745g for 20-25min, and the 2 O washed and precipitated 2 times, and the precipitate was dried at normal temperature to obtain the MSN nanoparticles.

[0029] (2) Preparation of MSN-DLDH nanoparticles: resuspend the MSN nanoparticles obtained in step (1) in ddH 2 In O, form the...

Embodiment 1

[0041] Embodiment 1: Preparation of oral nano-vaccine of large yellow croaker:

[0042] (1) Preparation of MSN nanoparticles: Add 4.8g of cetyltrimethyl-p-toluenesulfonate, 0.7mL of triethanolamine and 250mL of water into a round bottom flask, stir and dissolve at 80°C for 1h; then quickly add 10mL of silicic acid Tetraethyl ester, continue to stir and react for 2h; centrifuge at 12745g for 20min to recover the precipitate; use ddH for precipitation 2 Washed twice with O, followed by 6 g / L NH at 60 °C 4 NO 3 The methanol solution extracted the product 3 times; after the extraction, the precipitate was recovered by centrifugation at 12745g for 20min, and the 2 O washed and precipitated 2 times, and the precipitate was dried at normal temperature to obtain the MSN nanoparticles.

[0043] (2) Preparation of MSN-DLDH nanoparticles: resuspend the MSN nanoparticles obtained in step (1) in 20mL ddH 2 In O, the MSN nanoparticle suspension liquid that obtains concentration is 23.2m...

Embodiment 2

[0048] Embodiment two: the preparation of immune type large yellow croaker feed:

[0049] Resuspend the MSN-DLDH@HP55 enteric-coated nanoparticles obtained in step (3) of Example 1 in 200mL ddHO 2 In O, a suspension of MSN-DLDH@HP55 enteric nanoparticles with a concentration of 1.77mg / mL was formed;

[0050] MSN-DLDH@HP55 vaccine mix: Next, mix 2kg of large yellow croaker No. 2 feed with 3g of adhesive, and then evenly spray MSN-DLDH@HP55 enteric-coated nanoparticle suspension on the surface of large yellow croaker No. 2 feed Stir evenly again, spray 1mL of MSN-DLDH@HP55 enteric-coated nanoparticle suspension per 20g of large yellow croaker No. 2 feed, and then vacuum freeze-dry at -30°C to obtain feed with MSN-DLDH@HP55 attached. The feed with MSN-DLDH@HP55 was soaked in the chitosan solution for 3 seconds to form a thin coating (the chitosan solution should not cover the feed with MSN-DLDH@HP55 during soaking), and quickly transferred to ddH 2 Soak in O for 5s and take out...

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Abstract

The invention relates to an oral nano-vaccine of large yellow croaker and an immune type large yellow croaker feed prepared by the same. The oral nano-vaccine comprises MSN-DLDH nano-particles and enteric coatings coating the surfaces of the MSN-DLDH nano-particles. The MSN-DLDH nano particles are nano particles formed by assembling DLDH protein and a mesoporous silica micro-nano carrying system;and the enteric coatings are hydroxypropyl methylcellulose phthalate. The nano-vaccine disclosed by the invention has the advantages of low cost, simple preparation process, good capability of tolerating a strongly acidic stomach environment of fishes and the like. Besides, the vaccine has no toxicity to kidney cells of the large yellow croaker and is good in biocompatibility; and thus a technicalsupport is provided for research and development of oral nano-vaccines and clinical application in the future.

Description

technical field [0001] The invention relates to a large yellow croaker oral nano vaccine and its preparation method and application. Background technique [0002] Long-term irregular breeding, inbreeding, and nutritional imbalance have increased the risk of new and recurrent disease outbreaks in large yellow croakers. The main diseases that are prevalent in large yellow croakers include white gill disease, vibriosis, and iridescent virus disease. Among them, the diseases caused by bacteria, because of their high incidence, wide range of epidemics, and the most serious harm, pose a great threat to the aquaculture industry and food safety. It has been reported that large yellow croaker diseases caused by Vibrio parahaemolyticus, Vibrio alginolyticus and Vibrio harveyi have become the main restrictive factors for the sustainable development of large yellow croaker aquaculture. The use of fish vaccines will reduce the risk of fish infection with viruses, bacteria or parasites, ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K39/106A61K39/39A61K9/51A61K47/69A61K47/38A61P31/04
CPCA61K39/107A61K39/39A61K9/5161A61K47/6949A61P31/04A61K2039/55505A61K2039/542A61K2039/552
Inventor 吴允昆朱春华张维彬肖方南王前超张娜
Owner FUJIAN NORMAL UNIV
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