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CN-122005451-A - Vaccine based on angelica polysaccharide derivative delivery carrier, and preparation method and application thereof

CN122005451ACN 122005451 ACN122005451 ACN 122005451ACN-122005451-A

Abstract

The invention provides a vaccine based on a angelica polysaccharide derivative delivery carrier, and a preparation method and application thereof, and belongs to the technical field of biological medicines. The invention provides a preparation method of a vaccine, which uses amphiphilic angelica polysaccharide derivatives as delivery vehicles to obtain W/O/W type Pickering emulsion, namely the vaccine. The vaccine improves the safety of the vaccine and reduces the degradation of antigens in the digestive tract through the delivery of the angelica polysaccharide derivative. The invention further provides application of the vaccine in preparing fish culture related products and fish feed, wherein the vaccine is contained in feed particles, and the vaccine feed particles excite higher innate immunity and adaptive immunity levels of fish bodies and have good application prospects.

Inventors

  • MAO YUNXIANG
  • LI YANG
  • GAO YATING
  • TANG LEI
  • MO ZHAOLAN
  • YU XINZI

Assignees

  • 中国海洋大学三亚海洋研究院
  • 中国海洋大学

Dates

Publication Date
20260512
Application Date
20260414

Claims (10)

  1. 1. A method for preparing a vaccine based on a angelicapolysaccharide derivative delivery vehicle, comprising the steps of: (1) Dissolving stearic acid, 1-ethyl-3- (3-dimethylaminopropyl) carbodiimide hydrochloride and 4-dimethylaminopyridine in DMSO according to a molar ratio of 1:1:1.2, and performing an activation reaction to obtain a reaction solution, dropwise adding the reaction solution into a DMSO solution of angelica polysaccharide, reacting at room temperature, dropwise adding the reaction solution into absolute ethyl alcohol under an ultrasonic condition after the reaction is finished, centrifuging, collecting precipitate, dispersing the precipitate in water for dialysis, and freeze-drying after the dialysis is finished to obtain the angelica polysaccharide derivative; (2) Dispersing the angelica polysaccharide derivative in a sodium alginate solution to obtain an external water phase; (3) Dissolving antigen in PBS to obtain an internal water phase; (4) Mixing peanut oil and polyglycerol polyricinoleate to obtain an oil phase; (5) Mixing the internal water phase and the oil phase, and shearing to obtain colostrum; (6) Mixing and stirring the external water phase and the colostrum to obtain the W/O/W type pickering emulsion.
  2. 2. The method according to claim 1, wherein in the step (1), the final concentration of stearic acid in DMSO is 50mmol/L, and the activation reaction time is 30min.
  3. 3. The preparation method of claim 1, wherein in the step (1), the final concentration of angelica polysaccharide in DMSO solution is 10 mg/mL-50 mg/mL, and the reaction time at room temperature is 36h.
  4. 4. The preparation method of claim 1, wherein in the step (1), the ultrasonic power is 100-200W, the centrifugation condition is 8000rmp,10min, the dialysis molecular weight cutoff is 8000-14000, the time is 48h, the freeze-drying temperature is-50 ℃ to-80 ℃, the vacuum degree is 2-10 Pa, and the time is 24-48 h.
  5. 5. The preparation method of claim 1, wherein the concentration of the sodium alginate solution in the outer water phase is 0.5%, the concentration of the angelica polysaccharide derivative is 1g/100mL, the concentration of the antigen in the inner water phase is 50mg/mL, and the volume ratio of peanut oil to polyglycerol polyricinoleate in the oil phase is 10:1.
  6. 6. The method according to claim 1, wherein the volume ratio of the internal aqueous phase to the oil phase is 2:8, 3:7, 4:6 or 5:5, the shearing speed is 1000 rpm, and the time is 2min.
  7. 7. The method according to claim 1, wherein the volume ratio of the primary emulsion to the external aqueous phase is 4:6, 5:5, 6:4 or 8:2, and the stirring speed is 8000 rpm for 1min.
  8. 8. The vaccine prepared by the preparation method of any one of claims 1 to 7.
  9. 9. Use of the vaccine of claim 8 for the preparation of fish farming-related products.
  10. 10. A fish feed is characterized in that the vaccine of claim 8 is contained in feed particles, and the vaccine content is 0.1% -0.5% of the mass of the feed particles.

Description

Vaccine based on angelica polysaccharide derivative delivery carrier, and preparation method and application thereof Technical Field The invention belongs to the technical field of biological medicine, and particularly relates to a vaccine based on a angelica polysaccharide derivative delivery carrier, and a preparation method and application thereof. Background Aquaculture is an important part of agricultural economy, and in recent years, with development of ecological transformation of fishery, intensive cultivation modes such as indoor cultivation and industrial cultivation are becoming more and more mainstream. However, the problem of the following aquatic diseases is also increasingly prominent, and the aquaculture industry faces serious challenges of disease frequency. At present, the prevention and treatment of fish diseases also depends on chemical medicines to a great extent, and excessive use of the chemical medicines possibly causes problems in food and environment, hidden dangers such as antibiotics abuse and the like, and does not meet the modern aquaculture targets of high efficiency, safety and environmental friendliness. Immunization is one of the effective means of preventing and controlling fish diseases. The oral immunization is taken as an immunization way, has the characteristics of convenient use, small fish group stress, suitability for large-scale immunization and repeated immunization, and is especially suitable for the immunization of aquatic animals. At present, research shows that the pinocytosis capability of the intestinal cells of the rear segment of fish can absorb and transport soluble or granular antigens, can rapidly release the antigens to the cell gap, can be presented by macrophages in epithelium, and is a key effect part of the intestinal mucosa immunity of fish. However, due to the destruction of the fish gut environment and various digestive enzymes, vaccine antigen components have been largely degraded before reaching the hindgut mucosa, and small amounts of antigens reaching the gut mucosa are also difficult to efficiently recognize and capture by the immune system to induce the generation of high level mucosal immune responses. In order to protect the antigen component from the environment of the digestive tract, it is necessary to combine a vaccine with a specific oral administration system to maintain the stable presence of the antigen in the digestive tract, and to enhance the level of immune system response and increase the antigen utilization efficiency. The radix Angelicae sinensis polysaccharide (ANGELICA SINENSIS polysaccharide) is the main active ingredient of traditional Chinese medicine radix Angelicae sinensis (ANGELICA SINENSIS), and has the biological activities of enhancing immunity, resisting oxidation, resisting tumor, promoting hematopoiesis, etc. However, at present, angelica polysaccharide-based vaccines which are successfully developed and applied to aquatic animals are not yet developed, and the effect of angelica polysaccharide serving as a vaccine carrier in fish is not known. Disclosure of Invention In order to solve the problems in the prior art, a first object of the present invention is to provide a vaccine based on a delivery vehicle of angelica polysaccharide derivatives and a preparation method thereof. The second object of the invention is also to provide the use of the vaccine in the preparation of fish farming related products. The third object of the present invention is also to provide a fish feed, the feed pellet containing the vaccine therein. In order to achieve the above object, the present invention provides the following technical solutions: the invention provides a preparation method of a vaccine based on a angelica polysaccharide derivative delivery carrier, which comprises the following steps: (1) Dissolving stearic acid, 1-ethyl-3- (3-dimethylaminopropyl) carbodiimide hydrochloride and 4-dimethylaminopyridine in DMSO according to a molar ratio of 1:1:1.2, and performing an activation reaction to obtain a reaction solution, dropwise adding the reaction solution into a DMSO solution of angelica polysaccharide, reacting at room temperature, dropwise adding the reaction solution into absolute ethyl alcohol under an ultrasonic condition after the reaction is finished, centrifuging, collecting precipitate, dispersing the precipitate in water for dialysis, and freeze-drying after the dialysis is finished to obtain the angelica polysaccharide derivative; (2) Dispersing the angelica polysaccharide derivative in a sodium alginate solution to obtain an external water phase; (3) Dissolving antigen in PBS to obtain an internal water phase; (4) Mixing peanut oil and polyglycerol polyricinoleate to obtain an oil phase; (5) Mixing the internal water phase and the oil phase, and shearing to obtain colostrum; (6) Mixing and stirring the external water phase and the colostrum to obtain the W/O/W type pickering emulsion. Preferably, in th