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An IgY for a PA-MSHA bacterial strain, and preparation method and application thereof

A technology of bacterial strains and biological products, applied in the field of mannose-sensitive Pseudomonas aeruginosa strain IgY and its preparation, can solve the problem that IgY of egg-laying poultry does not have a broad-spectrum antibacterial effect and anti-tumor auxiliary effect, and achieve good activity and specificity sexual effect

Active Publication Date: 2012-11-07
SHENZHEN BROSTIGER BIO PHARMA
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0009] The technical problem to be solved by the present invention is to provide a PA-MSHA strain IgY and its preparation method and application in view of the defects that the existing egg-laying poultry IgY does not have a broad-spectrum antibacterial effect and anti-tumor auxiliary effect

Method used

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  • An IgY for a PA-MSHA bacterial strain, and preparation method and application thereof
  • An IgY for a PA-MSHA bacterial strain, and preparation method and application thereof
  • An IgY for a PA-MSHA bacterial strain, and preparation method and application thereof

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Embodiment 1

[0031] Embodiment 1: the method for obtaining the product of egg yolk immunoglobulin with broad-spectrum antibacterial and anticancer

[0032] The antigen of the PA-MSHA strain is obtained through the above process, and an immunized egg, such as a chicken egg, is obtained after injection. The following describes the preparation process of yolk immunoglobulin with broad-spectrum antibacterial and anticancer properties extracted from immunized eggs. The extraction and purification method of the egg yolk immunoglobulin is water dilution method, repeated freezing and thawing method or ammonium sulfate, sodium sulfate, PEG, dextran sulfate, sodium alginate or caprylic acid precipitation or leaching, extraction, concentration and membrane filtration. Methods The antibody is extracted from egg yolk, and then the purified antibody is prepared into any pharmaceutical dosage form including tablets, injections, oral liquids and sprays by adding corresponding auxiliary materials including...

Embodiment 2

[0040] Embodiment 2: ELISA method detects the potency of egg yolk immunoglobulin

[0041] A 96-well plate was coated with Pseudomonas aeruginosa antigen at 10 μg / ml, overnight at 4°C, washed three times with PBS of pH 7.4 the next day, blocked with 5% calf serum for 12 hours, and air-dried after washing. Dilute the purified egg yolk immunoglobulin (starting from 1:10, a total of 8 concentration groups) with sterile PBS, add 100 μl to each well, and incubate at 37°C for 1 hour; after washing and air-drying, add horseradish peroxidase-labeled sheep Anti-chicken immunoglobulin (HRP-IgG, concentration 1:10 5 ) 100pl, incubated at 37°C for 1h. O-phenylenediamine was developed at 37°C in the dark for 15 minutes, the reaction was terminated with 2 mol / L sulfuric acid, and the A450 value was read on a microplate reader. Set sterile PBS as blank control. The test results confirmed that the titer of the purified egg yolk immunoglobulin was 1:10 5 .

Embodiment 3

[0042] Embodiment 3: egg yolk immunoglobulin stability test

[0043] Egg yolk immunoglobulin was sterilized by filtration, packed into small test tubes at 10 mg / ml, and stored at -20°C, 4°C and room temperature respectively, and its potency was tested regularly. The experimental results show that the stability of egg yolk immunoglobulin can be maintained for at least 12 months when stored at -20°C; the stability can be maintained for at least 6 months when stored at 4°C; the stability can be maintained for at least 6 months when stored at room temperature. It can be stored for more than 2 months.

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Abstract

The invention relates to an IgY for a mannose-sensitive hemagglutination pilus strain of Pseudomonas aeruginosa (PA-MSHA) and a preparation method and application thereof, wherein the preparation method comprises the steps of: (S1) preparing an antigen of the PA-MSHA bacterial strain; (S2) immunizing egg-laying poultry by injecting said PA-MSHA antigen, and checking immunized eggs laid by the immunized poultry; and (S3) taking out yolk of the immunized eggs to extract the IgY for the PA-MSHA bacterial strain. According to the invention, PA-MSHA is used to immunize egg-laying poultry, and the IgY for the PA-MSHA bacterial strain is then extracted from the poultry eggs. The IgY for the PA-MSHA bacterial strain can be further made into health-care and biological products to prevent and cure broad-spectrum opportunistic infection floras such as escherichia coli, pseudomonas aeruginosa, pneumococcus, deformation coccus, and helicobacter pylori, and to play a role of auxiliary treatment effect in various cancers such as breast cancer, liver cancer, lymph cancer, lung cancer, bladder cancer and leukemia, especially suitable for active and passive immunotherapy for cancers.

Description

technical field [0001] The present invention relates to mannose-sensitive Pseudomonas aeruginosa, more specifically, to a mannose-sensitive Pseudomonas aeruginosa (PA-MSHA) strain IgY, a preparation method thereof, related health care products, biological products and applications. Background technique [0002] 1. Limitations in clinical treatment of tumors [0003] At present, the number of cancer deaths in the world is about 7 million people every year, and it is particularly noteworthy that 24% of them occur in China, and the lack of active and effective drugs for targeted treatment of malignant tumors is the main reason. Patients with malignant tumors generally release immunosuppressive factors much higher than normal levels in the body in the middle and advanced stages, inhibit cell activity and directly inhibit the secretion of various lymphokines, thereby reducing the body's cellular and humoral immunity. At present, the treatment of malignant tumors is still based o...

Claims

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Application Information

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IPC IPC(8): C07K16/12C07K16/02A23L1/29A61K39/40A61P31/04A61P37/04A23L33/00
CPCC12N1/20C07K16/1214C07K2317/11C12R1/385C07K16/44C07K2317/23A61P31/04A61P35/00A61P37/04
Inventor 祁振强卢超
Owner SHENZHEN BROSTIGER BIO PHARMA
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