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Immune globulin and its prepn and application

A technology of immunoglobulin and egg-laying hens, applied in the direction of immunoglobulin, immunoglobulin from eggs, chemical instruments and methods, etc., can solve the problems of short half-life, large dosage, limited application, etc., and achieve high cure rate , strong specificity and excellent effect

Inactive Publication Date: 2004-09-08
KUNMING BEIOUSIDA BIOLOGICAL SCI TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the current use of immune proteins without booster immunization is large and the therapeutic effect is not good, and the half-life is short
These disadvantages increase the production and treatment costs of immunoglobulins, limiting their clinical application

Method used

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  • Immune globulin and its prepn and application
  • Immune globulin and its prepn and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] Cultivation and collection of Enterovirus 70 and Coxsackievirus A24: Viruses were cultivated for 48-72 hours, and the viruses were harvested when the cytopathic rate reached 75%. The virus culture medium was repeatedly frozen and thawed three times, the cells were disrupted by ultrasonic intermittently, centrifuged at 3,500 rpm for 20 minutes, and the supernatant was collected; centrifuged at 35,000 rpm for 3 hours, the precipitate was collected, suspended in phosphate buffer (PBS), and an equal volume of sucrose was placed at the bottom. Centrifuge at 40,000 rpm for 3 hours to collect the precipitate. Concentrates were assayed for immunology, protein concentration and CPE.

Embodiment 2

[0032] Screening and immunization of laying hens: Inject three-month-old specific pathogen-free (SPF) laying hens with pentavalent vaccines such as anti-avian influenza and anti-fowl plague, and observe that there is no abnormality for two weeks. Injection of inactivated virus liquid coated with incomplete Freund's adjuvant emulsification (100TCID 50 ); booster immunization once every 2 weeks; regular detection of serum and egg yolk immunoglobulin titers.

Embodiment 3

[0034] Separation and purification of specific protein: 75% alcohol aseptic treatment on the surface of immune chicken eggs, homogenization treatment after removing egg white, adding sterile phosphate buffer solution of pH7.6 10mM in 1:4 for homogenization, 3.5% (V / W ) of PEG6000 was stirred and added, 10000 rpm for 20 minutes, and the supernatant was collected, 8.5% (V / W) PEG6000 was added with stirring, 10000 rpm for 20 minutes, and the precipitate was collected, and the pH7.6 10mM Phosphate buffer, dissolve the precipitate, add 12% (V / W) PEG6000 with stirring, centrifuge at 10,000 rpm for 20 minutes, collect the precipitate, and dissolve the precipitate with a small amount of pH 8.0 25mM phosphate buffer. Purified by DEAE-Sephacel column chromatography gel filtration, collected immunoglobulin peaks, concentrated ammonium sulfate and dialyzed at 4°C.

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Abstract

The present invention discloses one kind of immune globulin and its preparation and application. The immune globulin is type-70 enterovirus resisting and type-A24 Coxsachie virus resisting chicken yolk immune globulin and is prepared into product with antibody activity. The refined effective component has high purity and high activity, and is ideal product for eye drop and eye ointment. The immune globulin is used in preparing medicine for preventing and treating acute haemorrhagic conjunctivitis caused by type-70 enterovirus and type-A24 Coxsachie virus infection, and the medicine has high specificity, high curative rate and effect superior to conventional medicine.

Description

technical field [0001] The invention relates to an immunoglobulin and a preparation method thereof. At the same time, the present invention also relates to the use of the immunoglobulin in the field of pharmacy. Background technique [0002] Acute hemorrhagic conjunctivitis (AHC) is also called epidemic hemorrhagic conjunctivitis. It is mainly caused by enterovirus type 70 and coxsackie A24 mutant strain (coxsackie A24). It is a new type of conjunctivitis and keratitis. It became popular in Southeast Asia in 1969 and spread to my country in 1971. Children are more likely than adults Mild, infants and young children rarely suffer from the disease. It often occurs in summer and autumn and has high prevalence. It is one of the Class C infectious diseases managed by the Law of the People's Republic of China on the Prevention and Control of Infectious Diseases. The disease is characterized by a short incubation period, the shortest being only 2 to 3 ho...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K39/395A61P27/02A61P31/12C07K16/02C07K16/44
Inventor 朱毅果余红朱琪
Owner KUNMING BEIOUSIDA BIOLOGICAL SCI TECH
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