Preparation for specific aspergillus fumigatus infection resisting chicken egg yolk antibody and application

A technology of chicken yolk antibody and Aspergillus fumigatus, which is applied in the field of immunity, can solve the problems of patients' allergic reactions, repeated patients' illnesses, poor water solubility, etc., and achieve the effect of less allergic reactions, good water solubility, and uncomplicated production process

Inactive Publication Date: 2015-08-19
THE AFFILIATED HOSPITAL OF QINGDAO UNIV +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] Common antifungal drugs have disadvantages such as poor water solubility, high toxicity and side effects, and allergic reactions in some patients to varying degrees.
Many patients stop treatment due to intolerance, the curative effect is often poor, and the patient's condition often relapses. Currently, there is no drug that can effectively control fungal infection for a long time in the treatment of fungal diseases

Method used

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  • Preparation for specific aspergillus fumigatus infection resisting chicken egg yolk antibody and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] Preparation of chicken egg yolk antibody specific for Aspergillus fumigatus infection:

[0022] (1) Prepare inactivated Aspergillus fumigatus mycelia suspension: inoculate Aspergillus fumigatus and cultivate in Sabouraud liquid medium for 5 days, take out Aspergillus fumigatus mycelia mass and grind in a grinder, filter spores, and finally adjust the mycelium concentration to 2×10 9 CFU / ml;

[0023] (2) The mycelium concentration was 2×10 9 The CFU / mL bacterial suspension and Freund's complete adjuvant were mixed at a ratio of 1:1 and fully emulsified. Each chicken was injected (1 mL / bird), and after 3 weeks, 5 weeks, and 8 weeks, it was replaced with Freund's incomplete adjuvant. Booster immunization 3 times.

[0024] (3) Use the ammonium sulfate salting-out method to prepare specific chicken yolk antibody protein against Aspergillus fumigatus infection, break the eggs to remove the egg white and yolk film, mix the egg yolk liquid with ultrapure water at a ratio of...

Embodiment 2

[0027] In vitro antibacterial test of chicken egg yolk antibody specific against Aspergillus fumigatus infection:

[0028] After Aspergillus fumigatus was shaken and cultivated in the culture medium for 96 h, the concentration of Aspergillus fumigatus hyphae in the culture medium was calculated by blood cell counting method, which was converted into absorbance value by ultraviolet spectrophotometer to calculate the total amount of mycelium in the sample. Will grind 1×10 9 CFU / ml mycelia were serially diluted with Sabouraud liquid medium, the absorbance value of each dilution was measured at 600 nm, and a standard curve was drawn between mycelium concentration and absorbance value. Will 1×10 5 The CFU / ml bacterial suspension and the antibody with a titer higher than 1:10000 obtained in Example 1 were prepared into different concentrations of chicken egg yolk antibody, and the absorbance was measured and counted after 24h, 48h, and 72h of mixed culture. Depend on figure 1 It ...

Embodiment 3

[0030] Test of physicochemical properties of specific anti-Aspergillus fumigatus-infected chicken egg yolk antibody:

[0031] Dilute the IgY solution to a specific concentration (the protein concentration is 0.02mg / mL), incubate in water baths at 40, 50, 60, 70, 80, and 90°C for 30 minutes, and then measure the antibody activity by indirect ELISA; at the same time, the protein concentration is 0.02 mg / mL IgY solution was incubated at the critical temperature of its activity loss for 0.5, 1.0, 1.5, 2.0, 2.5, 3.0, 3.5, 4.0, 4.5 h, and the activity of the antibody was measured by indirect ELISA. When investigating the acid-base stability of the antibody, use PBS solutions with pH 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, and 14 to dilute the IgY solution to a protein concentration of 0.02 mg / ml, after standing at 4°C for 1 hour, the antibody activity was measured by indirect ELISA.

[0032] The test results show that the antibody titer obtained in Example 1 is higher than the ...

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Abstract

The invention relates to the technical field of immunity, in particular to a specific aspergillus fumigatus infection resisting chicken egg yolk antibody and application thereof. The specific aspergillus fumigatus infection resisting chicken egg yolk antibody is prepared by the following steps: 1, preparing an aspergillus fumigatus antigen; 2, performing immune injection on laying hens; 3, preparing specific chicken egg yolk antibody freeze-dried powder by utilizing an ammonium sulfate salting-out method. The valence of the prepared specific aspergillus fumigatus infection resisting chicken egg yolk antibody is higher than 1:10,000; the prepared specific aspergillus fumigatus infection resisting chicken egg yolk antibody is a biological agent, the side effects generated to the human body are small, the irritation to human body conjunctivas, corneas, mucosa, skin and the like is small, no adverse reaction exists when the antibody is externally used or orally taken, and the physicochemical property is stable; the antibody can be saved for a long time and is particularly suitable for being locally applied to treating aspergillus fumigatus infection of ocular surface, oral cavity, skin and the like.

Description

technical field [0001] The invention relates to the technical field of immunization, in particular to the preparation and application of a chicken egg yolk antibody specific for resisting Aspergillus fumigatus infection. Background technique [0002] In recent years, with the widespread application of antibacterial drugs, immunosuppressants, chemotherapy drugs and radiotherapy, as well as the application of invasive diagnostic and therapeutic techniques, the rate of opportunistic fungal infections has increased significantly. Aspergillus is an important opportunistic fungus, and Aspergillus fumigatus is the most common Aspergillus genus. Common drugs for the treatment of fungal diseases in my country include natamycin, amphotericin, and new triazoles. According to the report of the World Health Organization, fungal infection is one of the main causes of blindness in pulmonary infection and corneal infection worldwide, and Aspergillus infection in the lung is the main cause ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K16/14C07K16/02A61K39/395A61P27/02A61P31/10
Inventor 赵桂秋林静高昂徐强黄晓明王谦姜楠胡丽婷李翠
Owner THE AFFILIATED HOSPITAL OF QINGDAO UNIV
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