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Nanoemulsion vaccine adjuvant

A vaccine adjuvant and nanoemulsion technology, applied in the field of biomedicine, can solve problems such as high cost, long process operation time, 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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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] Preparation of nanoemulsion vaccine adjuvant

[0035]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.

[0036] 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 1 As...

Embodiment 2

[0041] Product Stability Test

[0042] 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.

[0043] Table 2 The particle size of nanoemulsion vaccine adjuvant for 12 months

[0044]

[0045]

Embodiment 3

[0047] Sensitization of vaccine adjuvants

[0048] 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 nanoemulsion vaccine adjuvant, and a preparation method and applications thereof. According to the preparation method, squalene is taken as an oil phase, and is mixed with two surfactants, and a buffer solution is added so as to obtain a water phase mixture, wherein HLB value of the whole system ranges from 8 to 12, the total amount of the surfactants ranges from 8 to 20%, and water / oil ratio ranges from 0.2 to 0.8; the water phase mixture is subjected to rapid mixing stirring for preliminary emulsion at a temperature lower than phase inversion temperature, and is subjected to preheating at a certain stirring speed for further emulsion; an obtained mixed material is heated to a temperature higher than phase inversion temperature and is cooled, so that the oil phase is coated by the water phase completely, and the nanoemulsion vaccine adjuvant with a uniform particle size is obtained. The preparation method of the nanoemulsion vaccine adjuvant is simple and convenient; no high energy input is needed; particle size distribution of the nanoemulsion vaccine adjuvant is uniform; stability is high; and immune synergy effect is excellent.

Description

technical field [0001] The invention belongs to the field of biomedicine and relates to a vaccine adjuvant, in particular to a novel nanoemulsion vaccine adjuvant and a preparation method thereof. Background technique [0002] 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 obviously. Practice has proved that the combined application of vaccines and adjuvants can not only significantly improve the immune effect of vaccines, but also reduce the immune tolerance of an...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K39/39
Inventor 潘皓谢小虎张岳智
Owner 潘皓
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