Process for preparing antibody for small molecular substance using nanoliposome immune adjuvant

A technology of small molecular substances and nano-liposomes, which is applied in the field of immunology, can solve the problems of difficulty in producing high-titer antibodies, instability, and susceptibility to interference from other factors, so as to increase stability, ensure immune effects, Good biocompatibility effect

Inactive Publication Date: 2009-10-28
NANCHANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, some small molecular substances are unstable or easily interfered by other factors after entering the animal body, and it is difficult to produce high-titer antibodies with traditional emulsified adjuvants

Method used

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  • Process for preparing antibody for small molecular substance using nanoliposome immune adjuvant
  • Process for preparing antibody for small molecular substance using nanoliposome immune adjuvant
  • Process for preparing antibody for small molecular substance using nanoliposome immune adjuvant

Examples

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Embodiment 1

[0025] Example 1. Preparation of anti-fluoroquinolone antibiotic maternal monoclonal monoclonal antibodies using liposomal immunization route.

[0026] as attached figure 2 As shown on the left, the box part is the mother nucleus of fluoroquinolone antibiotics, which is the common structure of dozens of fluoroquinolones. Obtaining antibodies against the mother nucleus of fluoroquinolone antibiotics can detect the total residues of fluoroquinolones in food. However, as attached figure 2 As shown on the right, the fluoroquinolone antibiotic core will chelate with metal ions in the body, and then bind to other proteins to be masked, making it difficult to successfully produce specific antibodies against the fluoroquinolone antibiotic core.

[0027] Using traditional Freund's adjuvant for immunization, the antigen solution should be mixed with Freund's adjuvant in equal amounts, fully vibrated and emulsified, and then subcutaneously injected into animals at multiple points. D...

Embodiment 2

[0030] Example 2. Preparation of anti-amoxycillin monoclonal antibodies using liposomal immunization route.

[0031] as attached image 3 As shown, amoxicillin reacts with the amino group on the carrier protein through the carboxyl group, and connects to the carrier protein to synthesize an artificial antigen to immunize animals, hoping to obtain antibodies against amoxicillin.

[0032]However, the β-lactam four-membered ring in the structure of amoxicillin is easy to hydrolyze and open the ring due to the large bond angle tension. Mix artificial antigen with traditional emulsified adjuvant and inject it subcutaneously into animals for immunization. It takes a long time to generate an immune response. Specific antibodies against amoxicillin.

[0033] Using the technology of the present invention, using the reverse evaporation method, the embedding material soybean lecithin and cholesterol are mixed at a weight ratio of 1.8 / 1, dissolved in dichloromethane solvent at a ratio ...

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Abstract

The invention relates to a process for preparing antibody for small molecular substance using nanoliposome immune adjuvant, which belongs to the field of immunologic technique, including mixing soybean lecithin and cholesterol according to a weight ratio of 1.8-2.5/1, dissolving the mixture into a methylene dichloride solvent with W/V at 1/500-800, adding into a flask, vacuum evaporating at a temperature from 35 to 40 DEG C. on a rotary evaporator; charging an antigen solution with concentration ranging from 1.5 to 2.0mg/mL, and ratio to the embedding material ranging from 2.0 to 2.5/1, strongly swinging, wherein 20mM PBS with pH ranging from 6.8 to 7.2 is used as buffer; intravenous immune is carried out at the amount of 200-300 mu g per mice on each time; secondary booster immune; selecting mouse with high special antibody potency for cell fusion experiment, screening needed monoclonal antibody. The invention has advantage that: (1) the antibody is great in solubility with the embedding material, easy to be dissolved in the animal body and non-toxic; (2) liposome is rapidly captured by an immune cells when introduced into peripheral blood and distributed to lymph nodes and central immune organs, and antigen presenting is realized; (3) the antigen is embedded into the liposome, so that stability of antigen is increased, and immune effect is ensured.

Description

technical field [0001] The patent of the invention belongs to the technical field of immunology, and specifically relates to a method for preparing antibodies against small molecule substances. Background technique [0002] Antigens are exogenous substances that can stimulate the animal body to produce an immune response. Immune adjuvants, also known as non-specific immune proliferators, are not antigenic in themselves, but injected into the body together with antigens can enhance the immune effect. The biological effects of immune adjuvants include: (1) changing the physical properties of the antigen, causing the antigen to be released slowly and prolonging the action time of the antigen; (2) increasing the surface area of ​​the antigen, making the antigen easy to be phagocytized by macrophages; ( 3) The adjuvant can stimulate phagocytes to process the antigen; (4) The adjuvant can promote the contact between lymphocytes and enhance the function of helper T cells. [0003...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P21/08C07K16/44
Inventor 许杨郭杰标何庆华李燕萍黄志兵
Owner NANCHANG UNIV
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