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A kind of sheet calcium phosphate vaccine adjuvant and preparation method thereof

A vaccine adjuvant, calcium phosphate technology, applied in the field of biomedicine preparation, nano-micro materials, to achieve the effect of simple preparation process, safe and easy-to-obtain materials, and enhanced package throughput

Active Publication Date: 2022-07-26
湖北汉江大健康产业有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The object of the present invention is to overcome the shortcoming of existing vaccine adjuvant, provide a kind of flake calcium phosphate vaccine adjuvant and preparation method thereof that can improve tumor immunotherapy effect

Method used

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  • A kind of sheet calcium phosphate vaccine adjuvant and preparation method thereof
  • A kind of sheet calcium phosphate vaccine adjuvant and preparation method thereof
  • A kind of sheet calcium phosphate vaccine adjuvant and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] Disperse the chitosan in water, adjust the pH with hydrochloric acid until the chitosan is completely dissolved, so that the final mass percentage concentration is 0.5%, and the concentration is 10 mg mL. -1 An ethanolic solution of 3,4-dihydroxyphenylpropionic acid and a concentration of 10 mg mL -1 The 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDC) aqueous solution was mixed uniformly in a ratio of 0.5:1, added to the chitosan solution under rapid stirring, and hydrogen The pH was adjusted to 5.5 with sodium oxide, reacted for 3 h, dialyzed in aqueous hydrochloric acid solution with pH 5.0 for 1 day, dialyzed with deionized water for 2 h, and freeze-dried to obtain catechol-modified chitosan (CSC);

[0027] In 3ml, 2mg / ml CSC aqueous solution, add 1.2mL PBS (0.2M PH=6) aqueous solution to the above, stir at 600r / min for 10min, then add 4.2mL, 100mM CaCl2 aqueous solution, stir for 10min, centrifuge, Wash twice with deionized water to obtain OVA-fre...

Embodiment 2

[0029] Disperse the chitosan in water, adjust the pH with hydrochloric acid until the chitosan is completely dissolved, so that the final mass percentage concentration is 3%, and the concentration is 30 mg mL -1 An ethanolic solution of 3,4-dihydroxyphenylpropionic acid and a concentration of 30 mg mL -1 The 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDC) aqueous solution was mixed uniformly in a ratio of 1:1, added to the chitosan solution under rapid stirring, and hydrogen The pH was adjusted to 5.5 with sodium oxide, reacted for 10 h, dialyzed in an aqueous hydrochloric acid solution of pH 6.0 for 3 days, dialyzed with deionized water for 8 h, and freeze-dried to obtain catechol-modified chitosan (CSC);

[0030] Add the same volume of OVA to 1.5ml, 4mg / ml of CSC aqueous solution, so that the final concentration of OVA is 1mg / ml, and stir until the mixture is uniform;

[0031] Add 1.2mL PBS (0.5M PH=6) aqueous solution to the above (1), stir at 600r / min fo...

Embodiment 3

[0034] Disperse the chitosan in water, adjust the pH with hydrochloric acid until the chitosan is completely dissolved, so that the final mass percentage concentration is 4%, and the concentration is 40 mg mL -1 An ethanolic solution of 3,4-dihydroxyphenylpropionic acid and a concentration of 40 mg mL -1 The 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDC) aqueous solution was mixed uniformly in a ratio of 2:1, added to the chitosan solution under rapid stirring, and then added to the chitosan solution with hydrogen peroxide. The pH was adjusted to 6.0 with sodium, reacted for 12 h, dialyzed in aqueous hydrochloric acid solution with pH 7.0 for 2 days, dialyzed with deionized water for 10 h, and freeze-dried to obtain catechol-modified chitosan (CSC);

[0035] Add the same volume of OVA to 1.5 ml, 8 mg / ml of CSC in water to make a final concentration of OVA of 5 mg mL -1 , stir until the mixture is uniform;

[0036] Add 4 mL of PBS (0.2M pH=7) aqueous soluti...

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Abstract

A flake calcium phosphate vaccine adjuvant and a preparation method thereof. The preparation method is as follows: (1) Dissolving a certain amount of chitosan in water, adjusting the pH with hydrochloric acid until the chitosan is completely dissolved, dissolving a certain amount of 3, 4 ‑Dihydroxyphenylpropionic acid and carbodiimide (EDC) were dissolved in ethanol and water, respectively, and the two were mixed well, then added to the chitosan solution under rapid stirring, and the pH was adjusted to about 5.5 with sodium hydroxide , reacted for a certain time, dialyzed, and freeze-dried to obtain catechol-modified chitosan (CSC). (2) Add the protein mimic antigen to the CSC aqueous solution and stir until it is evenly mixed; then, add the phosphate buffer solution, stir for a period of time, then add the CaCl2 aqueous solution, stir for the same time, centrifuge, and wash with water to obtain flaky phosphoric acid. Calcium vaccine adjuvant. The invention ensures the activity of the protein vaccine and the safety of using the adjuvant; the preparation process is simple, the raw materials are cheap, and the product is quick and easy to obtain.

Description

technical field [0001] The invention relates to a sheet calcium phosphate vaccine adjuvant and a preparation method thereof, belonging to the fields of nano-micro material and biological medicine preparation. Background technique [0002] An adjuvant is a substance added to a vaccine that can improve the immunogenicity of an antigen. It can induce a stronger immune response and reduce the dose and production cost of the vaccine for people who have adverse reactions to vaccine immunization. Polysaccharide adjuvants have attracted extensive attention in the preparation of vaccines and drugs due to their inherent immune regulation, biocompatibility, biodegradability, low toxicity, and safety. In addition, it has been confirmed that a variety of natural polysaccharides have good immune-promoting effects and can enhance the immune function of humoral, cellular and mucosal membranes, so polysaccharides have received extensive attention as immune adjuvants. [0003] Chitosan is a ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K39/39A61P37/04A61P35/00
Inventor 王晓莉张闯年裴萌月梁佳怡徐蓉马桂蕾孙洪范
Owner 湖北汉江大健康产业有限公司
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