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Preparation method for nanoemulsion vaccine auxiliary agent

A vaccine adjuvant and nanoemulsion technology, applied in the field of biomedicine, can solve the problems of long process operation time, high cost, and large energy demand

Inactive Publication Date: 2016-01-27
潘皓 +2
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Especially the influenza vaccine Fluad based on MF59 emulsion adjuvant It has been on the market for 10 years. This is a vaccine adjuvant prepared by ultra-high pressure homogenization. Long process operating times, high energy requirements and high costs

Method used

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  • Preparation method for nanoemulsion vaccine auxiliary agent
  • Preparation method for nanoemulsion vaccine auxiliary agent
  • Preparation method for nanoemulsion vaccine auxiliary agent

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Preparation of nanoemulsion vaccine adjuvant

[0032] 1) Coarse milk preparation: use squalene as oil phase, mix with Tween 80 and Span 80. In addition, PBS, a buffer agent with a neutral pH, was added as the aqueous phase. The HLB is 9.5, the total amount of surfactant is 11%, the ratio of water to oil is 0.4, stirred for 15-20 minutes and heated to 55-70°C. (2) Phase transition and formation of nanoemulsion: keep the temperature at 55-70°C, continue stirring for 5-10 minutes, then quickly transfer the hot emulsion to a large amount (5 times the initial total amount) of PBS buffer at 4-8°C Dilute in solvent while stirring constantly. Continue stirring for 10-30 minutes to prepare the nanoemulsion vaccine adjuvant product. Store the product in aliquots at 2-8°C in a refrigerator.

[0033] figure 1 It is a schematic diagram of temperature, particle size and conductivity in the preparation of nanoemulsion vaccine adjuvant PIT method in this embodiment, as figure 1As...

Embodiment 2

[0038] Product Stability Test

[0039] Stability is an important indicator of vaccine adjuvants. Table 2 shows the stability data of six batches of nanoemulsion vaccine adjuvants in Example 1 placed at 25°C for 12 months. As can be seen from Table 2, the nanoemulsion vaccine adjuvant has no significant change in particle size (the most important stability index) within 12 months, and the product quality is stable.

[0040] Table 2 The particle size of nanoemulsion vaccine adjuvant for 12 months

[0041]

Embodiment 3

[0043] Sensitization of vaccine adjuvants

[0044] Rabbit experiments are used to verify the sensitization of the vaccine adjuvant of the present invention. Wherein the control group is a conventional hemorrhagic fever vaccine without adjuvant (1:32 for type I antigen, 1:16 for type II), and the same hemorrhagic fever antigen is selected for the experimental group, and finally the vaccine adjuvant vaccine prepared in Example 1 is added. agent, the configuration process is the same as that of conventional non-adjuvanted vaccines. Type I antigen titers were 1:32, 1:16, 1:8, and type II antigen titers were 1:16, 1:8. A booster injection was given on the 7th day after the primary immunization, and blood was collected 21 days after the primary immunization for the determination of the neutralizing fluorescence titer. The result is as figure 2 As shown, it can be seen from it that the fluorescent antibody level after its immunization of the vaccine (type I and type II HA titer i...

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Abstract

The invention provides a preparation method for a nanoemulsion vaccine auxiliary agent. Squalene is employed as an oil phase and is mixed with two surfactants, and a buffer liquid is added as a water phase. In the whole system, the HLB value is 8-12, the surfactant sum scope is 8%-20%, and the ratio of water to oil is 0.2-0.8. The above mixture is mixed and stirred rapidly at a temperature lower than a phase transition temperature for primary emulsification; then preheating is carried out at a certain stirring speed for further emulsification; the mixture is heated to a temperature higher than the phase transition temperature, then cooling is carried out, the water phase coats the oil phase completely, and finally a nano-level vaccine auxiliary agent with uniform particles is formed. The preparation method is simple and fast, and needs no high energy input. In the prepared nano-level vaccine auxiliary agent, particles are distributed uniformly, the stability is excellent, and the immunization synergy is excellent.

Description

[0001] The patent application of the present invention is a divisional application of a Chinese patent application with an application date of June 18, 2014, an application number of 201410272698.2, and a nanoemulsion vaccine adjuvant. technical field [0002] The invention belongs to the field of biomedicine and relates to a preparation method of a novel nanoemulsion vaccine adjuvant. Background technique [0003] Vaccine adjuvants, also known as immune adjuvants (Adjuvant), are generally non-immunogenic when used alone, but when used in combination with antigenic substances, they can enhance the immunogenicity of antigenic substances, enhance the body's immune response, or change the type of body's immune response substance. Adjuvants are essentially a type of non-specific immune enhancer, which are usually inoculated before vaccine immunization alone or mixed with the vaccine at the same time. After the second immunization, the adjuvant can enhance the immunity more obvi...

Claims

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Application Information

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IPC IPC(8): A61K39/39A61P37/04
Inventor 潘皓谢小虎张岳智
Owner 潘皓
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