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Ultrasonic microvesicle as immuno adjuvant and vaccine carrier

A technology of ultrasonic microbubbles and vaccine carriers, applied in the field of biomedical engineering, can solve the problems that it is difficult to meet the needs of the development of new peptide vaccines and fail to stimulate immune responses, achieve safe repeatability, improve nucleic acid expression levels, and target good effect

Active Publication Date: 2009-10-07
许川山 +2
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Secondly, co-immunization of aluminum compounds with many recombinant or synthetic peptide vaccine antigens or nucleic acid (DNA) vaccine antigens failed to stimulate an effective immune response, making it difficult to meet the needs of new vaccine development for peptide vaccines and nucleic acid (DNA) vaccines

Method used

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  • Ultrasonic microvesicle as immuno adjuvant and vaccine carrier
  • Ultrasonic microvesicle as immuno adjuvant and vaccine carrier
  • Ultrasonic microvesicle as immuno adjuvant and vaccine carrier

Examples

Experimental program
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Effect test

example 1

[0017] Example 1: Preparation of Surfactant-like Microbubbles

[0018] Mix the surfactant Span60; Tween 80 in a ratio of 1:1, and at the same time use NaCl:KCl:Na 2 HPO 4 :KH 2 PO 4According to the mass ratio of 200:30:5:1, add 100ml of deionized water, and use NaOH to adjust the pH value of the mixture to 7.36, which is used as the medium; add 100ml of the medium into the mixture of Span60 and Tween 80 , the mixture obtained above was heated on a magnetic stirrer, the temperature was heated to 120 ° C, and the time was 10 minutes, and it was fully mixed to make it a milky suspension system; At the same time, add 0.6ml of perfluoropropane gas at a rate of 0.5ml / s; place the probe of the vibrometer at 0.5-2.0cm below the liquid surface for acoustic vibration treatment, and vibrate at 30% of the maximum output power for 3 minutes. The emulsified mixed solution after sonication is divided into 3 layers after standing for 35 minutes, and the middle layer is taken out and dilut...

example 2

[0019] Example 2: Preparation of Lipid Microvesicles

[0020] Lecithin, cholesterol, polyethylene glycol stearyl ethanolamine were dissolved in chloroform at a mass ratio of 1:3:3, and evaporated in a rotary vacuum to form a film; 0.9% sodium chloride solution, propylene glycol and glycerin (0.9% chlorine Sodium chloride solution: propylene glycol: glycerin = 8:1:1), shake to wash the membrane to form a liposome suspension. Freeze overnight. After thawing, vibrate for 80s with a sonicator at 30% of the maximum output power, and at the same time slowly fill in 0.6ml of perfluoropropane gas below through the tee tube to form lipid fluorocarbon microbubbles (such as figure 2 ).

example 3

[0021] Example 3: Preparation of polymer material-polylactic acid / glycolic acid (PLGA) microbubbles

[0022] Add 0.1g of camphor into 20ml of dichloromethane, stir fully to make it completely dissolved; add 1.0g of high molecular material polylactic acid / glycolic acid polymer (PLGA) into the above solution, stir until it is completely dissolved, and then add 5% After 3ml of ammonium chloride is added, vibrate for 40s with a vibrator at 30% of the maximum output power to form a milky white emulsion; add the emulsion to 3% polyvinyl alcohol and homogenize it for 5 minutes, then add 2% isopropanol solution, stirred with a magnetic stirrer at room temperature for 2-5 hours, centrifuged at a high speed for 5 minutes (3000-5000 rpm), repeated 3 times; added 5% mannitol and fully mixed into a milky white solution to obtain PLGA microbubbles (such as image 3 )

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Abstract

The invention belongs to the field of biomedical engineering, and more specifically, the invention relates to a new type of ultrasonic microbubble used as an immune adjuvant and vaccine carrier. Ultrasound microbubbles carrying vaccine antigens are synthesized by adhering the vaccine antigen material to the surface of the microbubbles and / or wrapping them in the microbubbles; when used, the microbubbles carrying the vaccine antigens are applied to the target tissue through the system and locally, and ultrasonic waves are used to destroy the target tissue. The microbubbles in the tissue area release vaccine antigen substances in a localized manner and enhance the immune activity of the antigens. It is expected to be developed into a new immune adjuvant and vaccine carrier, especially new adjuvants and carriers suitable for peptide vaccines, nucleic acid vaccines, viral and bacterial vaccines.

Description

technical field [0001] The invention belongs to the field of biomedical engineering, and more specifically, the invention relates to a novel ultrasonic microbubble used as an immune adjuvant and a vaccine carrier. Background technique [0002] Traditional vaccines using inactivated or attenuated pathogens, cell components (such as capsular polysaccharides) or inactive bacterial toxins can usually induce the body to produce neutralizing antibodies to prevent infection, and immune adjuvants play an important role in this process . The immune adjuvant is a substance that does not produce immunogenicity when administered alone, but induces mucosal and / or systemic immunity when combined with an antigen. The only adjuvants currently used in human vaccines on a large scale are aluminum compounds, the most important of which is Al(OH) 3 . There are two types of vaccines using aluminum compounds in modern times, one is aluminum compound precipitation vaccines; the other is aluminu...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K39/385A61K38/39A61K47/30A61P35/00
Inventor 许川山王志刚于廷和刘志君
Owner 许川山
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