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Novel water-soluble immunologic adjuvant and preparing method thereof

An immune adjuvant and water-soluble technology, applied in the field of new water-soluble immune adjuvant and its preparation, can solve the problems of animal inflammatory response, large animal damage, and destruction of antigen conformation, so as to reduce the difficulty of screening, save the immunization cycle, Conservation of the effect of steric conformation

Pending Publication Date: 2019-06-28
苏州博特龙免疫技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] (2) The above-mentioned Freund's adjuvant and incomplete Freund's adjuvant are oily adjuvants, which are mixed with water-soluble antigens, requiring a cumbersome emulsification process, accompanied by eversion of the hydrophobic groups of water-soluble proteins, which will destroy the conformation of the antigen;
[0007] (3) The overall immunization cycle time is 8-12 weeks, which is relatively long. In terms of antigen dosage, the dosage of mouse antigen is 50-100ug / needle, and the dosage of rabbit antigen is 0.5-1.0mg / needle. The damage to the animal is relatively large, causing an inflammatory reaction in the animal, which is not in line with animal welfare

Method used

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  • Novel water-soluble immunologic adjuvant and preparing method thereof

Examples

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Comparison scheme
Effect test

Embodiment 1

[0022] A novel water-soluble immune adjuvant prepared according to the following steps:

[0023] 1. Weigh raw materials: Weigh 0.20g potassium dihydrogen phosphate, 1.30g sodium dihydrogen phosphate, 5.0g sodium chloride and 0.1g potassium chloride respectively;

[0024] 2. Dissolve the weighed potassium dihydrogen phosphate, sodium dihydrogen phosphate, sodium chloride and potassium chloride in double distilled water, set the volume to 1L and stir to dissolve, then use hydrochloric acid to adjust the pH value to 7.2 to obtain PBS phosphate buffer;

[0025] 3. Add 0.4g Poly I:C polymyocytes, 0.2g polylysine and 0.3g lentinan to the PBS phosphate buffer solution obtained in the previous step, and stir for at least 10 minutes with a magnetic stirrer to obtain the new The water-soluble immune adjuvant is then filtered and sterilized through a 0.22um filter, and finally stored in a refrigerator at 4°C.

[0026] Using conventional Freund's adjuvant as a comparison, Freund's adjuv...

Embodiment 2

[0030] A novel water-soluble immune adjuvant prepared according to the following steps:

[0031] 1. Weigh raw materials: Weigh 0.27g potassium dihydrogen phosphate, 1.80g sodium dihydrogen phosphate, 15.0g sodium chloride and 0.3g potassium chloride respectively;

[0032] 2. Dissolve the weighed potassium dihydrogen phosphate, sodium dihydrogen phosphate, sodium chloride and potassium chloride in double distilled water, set the volume to 1L and stir to dissolve, then use hydrochloric acid to adjust the pH value to 7.4 to obtain PBS phosphate buffer;

[0033] 3. Add 0.6g Poly I:C polymyocytes, 0.5g polylysine and 0.7g lentinan to the PBS phosphate buffer solution obtained in the previous step, and stir for at least 10 minutes with a magnetic stirrer to obtain the new The water-soluble immune adjuvant is then filtered and sterilized through a 0.22um filter, and finally stored in a refrigerator at 4°C.

[0034] Using conventional Freund's adjuvant as a comparison, Freund's adju...

Embodiment 3

[0038] A novel water-soluble immune adjuvant prepared according to the following steps:

[0039] 1. Weigh raw materials: Weigh 0.24g potassium dihydrogen phosphate, 1.50g sodium dihydrogen phosphate, 10.0g sodium chloride and 0.2g potassium chloride respectively;

[0040]2. Dissolve the weighed potassium dihydrogen phosphate, sodium dihydrogen phosphate, sodium chloride and potassium chloride in double distilled water, set the volume to 1L and stir to dissolve, then use hydrochloric acid to adjust the pH value to 7.3 to obtain PBS phosphate buffer;

[0041] 3. Add 0.5g Poly I:C polymyocytes, 0.3g polylysine and 0.5g lentinan to the PBS phosphate buffer solution obtained in the previous step, and stir for at least 10 minutes with a magnetic stirrer to obtain the new The water-soluble immune adjuvant is then filtered and sterilized through a 0.22um filter, and finally stored in a refrigerator at 4°C.

[0042] Using conventional Freund's adjuvant as a comparison, Freund's adjuv...

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Abstract

The invention discloses a novel water-soluble immunologic adjuvant which comprises a PBS phosphate buffer, Poly I:C, polylysine and lentinan. The PBS phosphate buffer is composed of monopotassium phosphate, sodium dihydrogen phosphate, sodium chloride, potassium chloride and double distilled water. Each liter of PBS phosphate buffer is prepared from 0.20-0.27 g of monopotassium phosphate, 1.30-1.80 g of sodium dihydrogen phosphate, 5.0-15.0 g of sodium chloride and 0.1-0.3 g of potassium chloride and the balance double distilled water to reach the constant volume of 1 L. In this way, a Bacillus Calmette Guerin vaccine does not need to be used, an animal body is prevented from generating antibodies against the adjuvant, and the purity of the antibodies can be easily improved; in addition, an animal can be immunized after the adjuvant is evenly mixed with water-based antigens, no complicated emulsifying process is needed, the spatial conformation of the antigens is effectively protected,the antibodies for the spatial conformation epitope can be more easily obtained, the immune cycle is effectively prolonged, the antigen consumption in each injection is effectively reduced, no harm is caused to the animal, and the animal welfare is better satisfied.

Description

technical field [0001] The invention relates to a novel water-soluble immune adjuvant and a preparation method thereof. Background technique [0002] Antigen is an exogenous substance that can stimulate the animal body to produce an immune response. The immune adjuvant itself does not have antigenicity, but it is applied before the antigen or at the same time as the antigen, which can non-specifically enhance or change the specific immune response of the body against the antigen. The ability to enhance the immunogenicity of the corresponding antigen. [0003] The adjuvant commonly used in the existing antibody preparation technology is Freund's adjuvant, and Freund's adjuvant is made up of mineral matter (paraffin oil), emulsifier (lanolin) and Bacillus Calmette-Guerin; It is mixed with lanolin, and the component ratio is 1-5:1, usually 2:1. Freund's complete adjuvant is based on the addition of BCG on the basis of incomplete adjuvant, and the final concentration is 2-20mg / ...

Claims

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Application Information

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IPC IPC(8): A61K39/39
Inventor 刘立成
Owner 苏州博特龙免疫技术有限公司
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