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Anti-echinococcosis antigen monoclonal antibody hybridoma cell strain, anti-echinococcosis antigen monoclonal antibody and application thereof

A hybridoma cell line and monoclonal antibody technology, applied in the fields of hybrid cell preparation, application, botanical equipment and methods, etc., can solve the problems of no simple and effective diagnostic method for human and canine early diagnosis, and the diagnosis method needs to be broken through.

Active Publication Date: 2020-06-12
VETERINARY INST XINJINAG ACADEMY OF ANIMAL SCI CLINIC MEDICAL SCI RES CENT XINJIANG ACADEMY OF ANIMAL HUSBANDRY SCI
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Problems solved by technology

From the point of view of the prevention and treatment of echinococcosis, there are two problems: (1) the final host has no vaccine for immunization; (2) there is no simple and effective diagnostic method for human and dog in the vast agricultural and pastoral areas. Early diagnosis, diagnostic methods need breakthroughs to meet the needs of large-scale census

Method used

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  • Anti-echinococcosis antigen monoclonal antibody hybridoma cell strain, anti-echinococcosis antigen monoclonal antibody and application thereof
  • Anti-echinococcosis antigen monoclonal antibody hybridoma cell strain, anti-echinococcosis antigen monoclonal antibody and application thereof
  • Anti-echinococcosis antigen monoclonal antibody hybridoma cell strain, anti-echinococcosis antigen monoclonal antibody and application thereof

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Experimental program
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Effect test

Embodiment

[0034] Embodiment: Before producing the monoclonal antibody and the hybridoma cell line secreting the monoclonal antibody, the following preparations need to be made:

[0035] 1. Preparation of experimental reagents: Triton X-100, bovine serum albumin (BSA), protein assayconcertration, protein marker, polyethylene glycol (PEG1500), RPMI-1640, fetal bovine serum purchased from Hyclone Company, 100×HT, 100×HAT was purchased from Hyclone Company, other reagents were of domestic analytical grade, low molecular mass standard protein marker was purchased from Promega Company; Tween-20, ammonium persulfate (APS), tetramethyldiethylamine (TEMED), 20× PBS concentrate bovine serum albumin (BSA), o-phenylenediamine (OPD), horseradish peroxidase-labeled goat anti-mouse IgG (HRP-IgG), dimethyl sulfoxide (DMSO), ELISA coating solution (pH7 .4), 8-azaguanine, and serum diluents are self-made by the laboratory; the rest of the reagents are domestic analytically pure;

[0036] 2. Preparation ...

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Abstract

The invention relates to an anti-echinococcosis antigen monoclonal antibody hybridoma cell strain, an anti-echinococcosis antigen monoclonal antibody and application of the anti-echinococcosis antigenmonoclonal antibody. The anti-echinococcosis antigen monoclonal antibody hybridoma cell strain is classified and named as a hybridoma cell strain XJ10D8D10. The strain is preserved in the China Center for Type Culture Collection (CCTCC) on September 25th, 2017. The preservation address is Wuhan University, Wuhan, China, and the preservation number is CCTCC NO: C201789. According to the invention,an echinococcosis adult surface membrane antigen is extracted; a Balb / c mouse is immunized; splenic lymphocytes secreting anti-echinococcosis adult surface membrane antigen are obtained; the hybridoma cell strain secreting the anti-echinococcosis adult surface film antigen is obtained by fusing the splenic lymphocytes with SP2 / 0 cells, so that an echinococcosis-resistant adult surface film antigen antibody is obtained, and the echinococcus granulosus infection of dogs can be sensitively detected through fecal antigen detecting by using the antibody as a diagnostic reagent. And necessary conditions are provided for preparation of large-scale general survey detecting kits.

Description

technical field [0001] This application relates to a monoclonal antibody against surface membrane antigen of adult Echinococcus granulosus and a hybridoma cell line that secretes the monoclonal antibody; this application also relates to a hybridoma cell line that produces a monoclonal antibody and secretes the monoclonal antibody Methods. Background technique [0002] Hydatid disease (echinococcosis) is a zoonotic parasitic disease caused by Echinococcus granulosus, which is prevalent worldwide, and the epidemic is becoming more and more serious. Highly prevalent in South America. Our country is also the hardest hit area. Immunodiagnosis, chemotherapy, diagnosis and vaccine of the final host of the disease are urgent problems to be solved and the main direction of in-depth research in the prevention and control of echinococcosis. Echinococcosis is distributed in 23 provinces and regions in my country, and seven provinces and regions including Xinjiang, Tibet, Gansu, Qingh...

Claims

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Application Information

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IPC IPC(8): C12N5/20C12N15/08C07K16/18G01N33/577C12R1/91
CPCC07K16/18C12N15/02G01N33/577G01N2333/43543Y02A50/30
Inventor 米晓云古努尔·吐尔逊
Owner VETERINARY INST XINJINAG ACADEMY OF ANIMAL SCI CLINIC MEDICAL SCI RES CENT XINJIANG ACADEMY OF ANIMAL HUSBANDRY SCI
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