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Vaccine adjuvant containing protamine and preparation method of vaccine adjuvant

A protamine and vaccine adjuvant technology, applied in the medical and health field, can solve the problems of difficult injection of mineral oil, difficult absorption, inflammation in the immune site, etc., so as to improve cellular and humoral immune responses, improve the immune effect of vaccines, and enhance strong immunity. effect of action

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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Mineral oil is difficult to inject and not easy to absorb, which can cause inflammation, necrosis and pain in the immune site
At the same time, the accumulation of residual carcinogenic components (polycyclic aromatic hydrocarbons) in mineral oil in animals seriously threatens food safety.
Aluminum adjuvant mainly induces Th2 type immune response, it is only suitable for vaccines with antibodies as the main protective immunity, and has no adjuvant effect on many viruses, parasites, and bacterial antigens
In addition, aluminum glue will form granulomas at the injection site and easily induce allergic reactions in the body

Method used

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  • Vaccine adjuvant containing protamine and preparation method of vaccine adjuvant
  • Vaccine adjuvant containing protamine and preparation method of vaccine adjuvant
  • Vaccine adjuvant containing protamine and preparation method of vaccine adjuvant

Examples

Experimental program
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Effect test

preparation example Construction

[0034] Such as Figure 7 As shown, in another embodiment of the present invention, a method for preparing a protamine-containing vaccine adjuvant is provided, the method comprising the steps of:

[0035] The preparation steps of the protamine solution: take a certain amount of protamine and fully dissolve it in normal saline for injection to prepare a solution with a concentration of 0.1-10%, filter and sterilize it through a filter membrane, and store it for later use;

[0036] Preparation steps of chitosan solution: Weigh a certain amount of chitosan and fully dissolve it in physiological saline for injection containing 0.2% acetic acid to prepare a solution with a concentration of 0.1-10%, adjust the pH to 4-6.5, and filter Membrane filtration, save for later use;

[0037] Preparation steps of thimerosal sodium salt solution: Weigh a certain amount of thimerosal sodium salt and fully dissolve it in physiological saline for injection to prepare a solution with a concentrati...

Embodiment 1

[0049] Embodiment 1: Screening research of adjuvant formula

[0050] 1. Materials and methods

[0051] 1. Reagents

[0052] Chicken ovalbumin (OVA) and protamine were purchased from sigma company; chitosan was purchased from aladdin company; thimerosal was purchased from Sangon Bioengineering (Shanghai) Co., Ltd.; sodium chloride injection was purchased from Fuzhou Haiwang Fuyao Pharmaceutical Co., Ltd.; HRP-labeled goat anti-mouse IgG antibody was purchased from SouthernBiotech.

[0053] 2. Preparation of adjuvant solution

[0054] Preparation of protamine solution: Weigh a certain amount of protamine and fully dissolve it in normal saline for injection to make a solution with a concentration of 1%, filter it through a 0.22 μm filter membrane, and store it at 4°C for later use.

[0055] Preparation steps of chitosan solution: Weigh a certain amount of chitosan and fully dissolve it in physiological saline for injection containing 0.2% acetic acid to make a solution with a ...

Embodiment 2

[0083] Example 2: Research on the synergistic adjuvant effect of protamine, chitosan and thimerosal

[0084] 1. Materials and methods

[0085] 1. Reagents

[0086] 20% aluminum gel adjuvant was donated by China Mugong Group Co., Ltd.; HRP-labeled goat anti-mouse IgM and IgG antibodies were purchased from SouthernBiotech; other reagents were the same as in Example 1.

[0087] 2. Preparation of adjuvant solution

[0088] The preparation steps of 1% protamine solution, 1% chitosan solution, and 0.1% thimerosal sodium salt solution are the same as in Example 1.

[0089] Preparation of 6 kinds of adjuvant solutions: 6 kinds of adjuvant solutions were prepared by using 20% ​​aluminum gel adjuvant and the above-mentioned protamine solution, chitosan solution and thimerosal solution as mother solutions. Solution 1: protamine content is 0.05%; solution 2: chitosan content is 0.1%; solution 3: protamine content is 0.05%, chitosan content is 0.1%; solution 4: thimerosal content is 0.0...

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Abstract

The invention discloses a vaccine adjuvant containing protamine, and relates to the field of vaccine adjuvants. The vaccine adjuvant comprises, by weight, 0.025-0.1% of protamine, 0.05-0.2% of chitosan, 0.00025-0.001% of thiomersalate and the balance normal saline. Meanwhile, the invention further provides a preparation method of the vaccine adjuvant containing the protamine. The method includes the steps that protamine and thiomersalate are dissolved in the normal saline, the chitosan is dissolved in the normal saline containing 0.2% acetic acid; the mixture is filtered through a 0.22-micronfiltering membrane, and the obtained sterile solution is an adjuvant solution containing the protamine. The adjuvant containing protamine, chitosan and thiomersalate is combined with a vaccine for animal immunization so that the immune response of animals to the vaccine and the immune effect of the vaccine can be greatly improved; one of the main components, namely thiomersalate, of the adjuvant has a corrosion-resisntat effect so that the adjuvant can be stored for a long time.

Description

technical field [0001] The invention relates to the medical and health field, in particular to a protamine-containing vaccine adjuvant and a preparation method thereof. Background technique [0002] The pressure of epidemic prevention and control in large-scale farms in my country is severe, and compulsory vaccination with vaccines is one of the effective measures to control animal infectious diseases. Practice has proved that the vaccine immunization effect of many animal diseases (PRRS, pseudorabies, foot-and-mouth disease, E. Immunocompromised groups caused by factors such as feeding, transfer, injection, etc. [0003] Protamine protein (PP) is a polycationic peptide rich in basic amino acids. It has antithrombotic, antitumor, liver function enhancement, and antibacterial effects. It has important applications in the fields of food, medicine, and health care. PP has a large number of positive charges, and can be closely combined with negatively charged DNA, heparin sodi...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K39/39A61P37/04A61K39/245A61P31/22A61K39/108A61P31/04
CPCA61K39/0258A61K39/12A61K39/39A61K2039/55511A61K2039/55516A61K2039/55583A61P31/04A61P31/22A61P37/04C12N2710/16734
Inventor 林炜明倪敬轩郑新添