A novel universal matrix vaccine adjuvant and its preparation method and use

Abstract

The invention relates to a novel universal matrix vaccine adjuvant and its preparation method and application. The adjuvant is a saponin matrix adjuvant, which contains abibis saponin, cholesterol and lecithin, and the weight ratio of the three is 5-20:1:1, and is composed of cubes and / or spherical particles with a particle diameter of 10-12 nm 300-700 nm sheet-like and / or chain-like particles. Albizia juniperin matrix adjuvant has a significant immune adjuvant effect, can enhance the cellular immunity and humoral immune response of the vaccine immune body, induce a balanced Th1 type and Th2 type immune response; and can reduce the dose of vaccine antigen, which is a limited amount of antigen Or it is an ideal adjuvant for expensive vaccines. Compared with Albizia Julibrissin, it has less hemolytic property and better safety. Albizia juniper saponin matrix adjuvant has good affinity with various antigens, and is a new type of universal vaccine adjuvant. The production process is simple and easy for large-scale production.

Description

technical field

[0001] The invention relates to a novel universal matrix vaccine adjuvant and its preparation method and application. Background technique

[0002] Vaccination is the most economical and effective means to prevent and control infectious diseases. New vaccines such as DNA vaccines, recombinant vaccines, and synthetic peptide vaccines are a type of biological preparations that have developed rapidly in recent years. Compared with conventional vaccines, they have the advantages of high antigen purity, strong specificity, and good safety. However, these novel vaccines have weak immunogenicity and low immune potency [Oyston P, Robinso K. The current challenges for vaccine development. J Med Microbiol2012; 61(7): 889–894]. At the same time, current vaccine research is facing important challenges from some complex pathogens such as malaria, tuberculosis, and human immunodeficiency virus [Leroux–Roels G. Unmet needs in modern vaccination: Adjuvants to improve the im...

Examples

Embodiment 1

[0034] Example 1: Preparation of Albizia Julibis Saponin Matrix Adjuvant (ASMA) by Dialysis

[0035] 1. Preparation of 5% Albizia Julibis Saponin Solution: Dissolve Albizia Julibis Saponin in distilled water to make a 50 mg / ml solution, filter and sterilize, and set aside.

[0036] 2. Preparation of 1% cholesterol / lecithin solution: take non-ionic surfactant Mega-10, dissolve in double distilled water to make a 20% solution; then weigh lecithin and cholesterol, dissolve in 20% Mega- 10 aqueous solution, make each ml contain 10 mg of lecithin and 10 mg of cholesterol, filter and sterilize, and set aside.

[0037] 3. Orthogonal experimental design: using L 9 (3 4 ) orthogonal table to investigate the prescription and process conditions. The factors and level design of the orthogonal experiment are shown in Table 1, and the experimental arrangement is shown in Table 2.

[0038] Table 1 Orthogonal test factor level table

[0039]

[0040] Table 2 Orthogonal test arrangemen...

Embodiment 2

[0046] Example 2: Preparation of Albizabis saponin matrix adjuvant (ASMA) by ultrafiltration

[0047] 1. Preparation of 5% Albizia Julibis Saponin Solution: Dissolve Albizia Julibis Saponin in distilled water to make a 50 mg / ml solution, filter and sterilize, and set aside.

[0048] 2. Preparation of 1% cholesterol / lecithin solution: take non-ionic surfactant Mega-10, dissolve in double distilled water to make a 20% solution; then weigh lecithin and cholesterol, dissolve in 20% Mega- 10 aqueous solution, make each ml contain 10 mg of lecithin and 10 mg of cholesterol, filter and sterilize, and set aside.

[0049] 3. Orthogonal experimental design: using L 9 (3 4 ) orthogonal table to investigate the prescription and process conditions. The factors and level design of the orthogonal experiment are shown in Table 1, and the experimental arrangement is shown in Table 2.

[0050] 4. Sample preparation is enlarged 5 times according to the volume in Table 3, add 1% cholesterol / l...

Embodiment 3

[0052] Example 3: Preparation of Albizia Julibrissin Matrix Adjuvant (ASMA) from Lecithin from Different Sources

[0053] 1. Preparation of 5% Albizia Julibis Saponin Solution: Dissolve Albizia Julibis Saponin in distilled water to make a 50 mg / ml solution, filter and sterilize, and set aside.

[0054] 2. Preparation of 1% cholesterol / lecithin solution: prepare a 20% aqueous solution with non-ionic surfactant Mega-10 or octyl glucoside. Then cholesterol and phosphatidylcholine, soybean lecithin or egg yolk lecithin were dissolved in 20% Mega-10 aqueous solution to obtain three groups of samples, containing 10 mg / ml each of phosphatidylcholine and cholesterol, and soybean lecithin and cholesterol 10 mg / ml each, or yolk lecithin and cholesterol 10 mg / ml each, sterilized by filtration and set aside.

[0055] 3. Sample preparation Take 0.3 ml each of the 1% cholesterol / lecithin solutions prepared from the above three different sources of lecithin, add 0.6 ml of PBS buffer solutio...

technical field

[0001] The invention relates to a novel universal matrix vaccine adjuvant and its preparation method and application. Background technique

[0002] Vaccination is the most economical and effective means to prevent and control infectious diseases. New vaccines such as DNA vaccines, recombinant vaccines, and synthetic peptide vaccines are a type of biological preparations that have developed rapidly in recent years. Compared with conventional vaccines, they have the advantages of high antigen purity, strong specificity, and good safety. However, these novel vaccines have weak immunogenicity and low immune potency [Oyston P, Robinso K. The current challenges for vaccine development. J Med Microbiol2012; 61(7): 889–894]. At the same time, current vaccine research is facing important challenges from some complex pathogens such as malaria, tuberculosis, and human immunodeficiency virus [Leroux–Roels G. Unmet needs in modern vaccination: Adjuvants to improve the im...

Examples

Embodiment 1

[0034] Example 1: Preparation of Albizia Julibis Saponin Matrix Adjuvant (ASMA) by Dialysis

[0035] 1. Preparation of 5% Albizia Julibis Saponin Solution: Dissolve Albizia Julibis Saponin in distilled water to make a 50 mg / ml solution, filter and sterilize, and set aside.

[0036] 2. Preparation of 1% cholesterol / lecithin solution: take non-ionic surfactant Mega-10, dissolve in double distilled water to make a 20% solution; then weigh lecithin and cholesterol, dissolve in 20% Mega- 10 aqueous solution, make each ml contain 10 mg of lecithin and 10 mg of cholesterol, filter and sterilize, and set aside.

[0037] 3. Orthogonal experimental design: using L 9 (3 4 ) orthogonal table to investigate the prescription and process conditions. The factors and level design of the orthogonal experiment are shown in Table 1, and the experimental arrangement is shown in Table 2.

[0038] Table 1 Orthogonal test factor level table

[0039]

[0040] Table 2 Orthogonal test arrangemen...

Embodiment 2

[0046] Example 2: Preparation of Albizabis saponin matrix adjuvant (ASMA) by ultrafiltration

[0047] 1. Preparation of 5% Albizia Julibis Saponin Solution: Dissolve Albizia Julibis Saponin in distilled water to make a 50 mg / ml solution, filter and sterilize, and set aside.

[0048] 2. Preparation of 1% cholesterol / lecithin solution: take non-ionic surfactant Mega-10, dissolve in double distilled water to make a 20% solution; then weigh lecithin and cholesterol, dissolve in 20% Mega- 10 aqueous solution, make each ml contain 10 mg of lecithin and 10 mg of cholesterol, filter and sterilize, and set aside.

[0049] 3. Orthogonal experimental design: using L 9 (3 4 ) orthogonal table to investigate the prescription and process conditions. The factors and level design of the orthogonal experiment are shown in Table 1, and the experimental arrangement is shown in Table 2.

[0050] 4. Sample preparation is enlarged 5 times according to the volume in Table 3, add 1% cholesterol / l...

Embodiment 3

[0052] Example 3: Preparation of Albizia Julibrissin Matrix Adjuvant (ASMA) from Lecithin from Different Sources

[0053] 1. Preparation of 5% Albizia Julibis Saponin Solution: Dissolve Albizia Julibis Saponin in distilled water to make a 50 mg / ml solution, filter and sterilize, and set aside.

[0054] 2. Preparation of 1% cholesterol / lecithin solution: prepare a 20% aqueous solution with non-ionic surfactant Mega-10 or octyl glucoside. Then cholesterol and phosphatidylcholine, soybean lecithin or egg yolk lecithin were dissolved in 20% Mega-10 aqueous solution to obtain three groups of samples, containing 10 mg / ml each of phosphatidylcholine and cholesterol, and soybean lecithin and cholesterol 10 mg / ml each, or yolk lecithin and cholesterol 10 mg / ml each, sterilized by filtration and set aside.

[0055] 3. Sample preparation Take 0.3 ml each of the 1% cholesterol / lecithin solutions prepared from the above three different sources of lecithin, add 0.6 ml of PBS buffer solutio...

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