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Anti-schistosoma japonicum Sjp40 chicken egg-yolk antibodies, and preparation method and application thereof

A chicken egg yolk antibody, schistosome technology, applied in chemical instruments and methods, from egg immunoglobulins, instruments and other directions, can solve the problems that have not been reported in research, and achieve the effects of simple method, high sensitivity and strong specificity

Inactive Publication Date: 2014-02-05
INST OF HYGIENE & ENVIRONMENTAL MEDICINE PLA ACAD OF MILITARY MEDICAL
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  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] Medical workers have done a lot of research on the detection of schistosomiasis japonicum, and some progress has been made, but there is no report on the use of IgY as the Sjp40 antigen of schistosomiasis japonicum

Method used

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  • Anti-schistosoma japonicum Sjp40 chicken egg-yolk antibodies, and preparation method and application thereof
  • Anti-schistosoma japonicum Sjp40 chicken egg-yolk antibodies, and preparation method and application thereof
  • Anti-schistosoma japonicum Sjp40 chicken egg-yolk antibodies, and preparation method and application thereof

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Embodiment 1

[0038] 1 Materials and methods

[0039] 1.1 The immunization of SPF chickens and the collection of recombinant Sjp40 immunized 22-week-old SPF grade Laihang laying hens (Beijing Experimental Animal Center), 0.1mg each time, the first immunization will be mixed with the same amount of Freund's complete adjuvant (SIGMA company in the United States), using Subcutaneous multi-point intramuscular injection. Then booster immunization was carried out at 5W, 6W, and 7W. Freund's incomplete adjuvant was used for immunization. Eggs after each immunization were collected and stored at 4°C to prepare egg yolk antibodies. 1.2 Extraction and purification of IgY IgY in egg yolk was separated and purified by water-diluted ammonium sulfate precipitation. Dilute egg yolk and distilled water at 1:8, adjust pH to 4.2-4.4 with hydrochloric acid, stir evenly at 4°C and let stand overnight, then centrifuge at 12000r / min for 30min at 4°C to collect the supernatant to obtain the water-soluble fractio...

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Abstract

The invention provides an anti-schistosoma japonicum Sjp40 chicken egg-yolk antibody which can be specifically bound with schistosoma japonicum Sjp40. The anti-schistosoma japonicum Sjp40 chicken egg-yolk antibody is capable of identifying the schistosoma japonicum Sjp40 through antigen specific binding activity detection thereof; the OD value of the anti-schistosoma japonicum Sjp40 chicken egg-yolk antibody is decreased as the antibody dilution is reduced; the anti-schistosoma japonicum Sjp40 chicken egg-yolk antibody has dependence on antigen concentration; the antibody is obtained by a method comprising the following steps of: (1) collecting eggs of healthy SPF (Specific pathogen Free) grade laying hen which is primarily immunized with purified Sjp40 antigen previously, and primarily extracting IgY from the eggs by the method of diluting ammonium sulfate precipitate with water; (2) further purifying the IgY orderly through a dialysis bag and a DEAE-FF (Diethylaminoethyl-Fast Flow) anion exchange column; (3) performing analysis and identification on the IgY through polyacrylamide gel electrophoresis, Western blotting and ELISA (Enzyme Linked Immunosorbent Assay), wherein the antibody is a chicken egg-yolk immune globulin (IgY) special for the schistosoma japonicum Sjp40; and therefore, a new idea is provided for early diagnosis and prevention and treatment of the schistosoma japonicum.

Description

technical field [0001] The invention relates to the field of biological technology, in particular to an anti-schistosoma japonicum Sjp40 chicken egg yolk antibody and a preparation method and application thereof. Background technique [0002] Schistosomiasis is a disease caused by tropical parasites. It is an important zoonotic disease. At present, 800 million people in the world are threatened, 200 million people are infected, and about 2.8 million people die from the disease every year. my country is an endemic area of ​​schistosomiasis and is one of the countries with the most serious schistosomiasis hazards in the world. The main hazard of schistosomiasis is the granulomatous lesions caused by the soluble egg antigen (SEA) secreted by the eggs. If early diagnosis can be realized, patients and livestock can be diagnosed and treated in time, which is of great significance for blocking the occurrence and development of serious lesions. [0003] Studies have shown that P38...

Claims

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Application Information

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IPC IPC(8): C07K16/20C07K16/02G01N33/569
Inventor 赵小玲陈伟强闫俊王俪蒙王尚王涛王军孙思明
Owner INST OF HYGIENE & ENVIRONMENTAL MEDICINE PLA ACAD OF MILITARY MEDICAL
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