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Hybridoma cell strain secreting high-neutralization activity infectious bursal disease virus (IBDV) monoclonal antibody

A monoclonal antibody, hybridoma cell technology, applied in the field of antibody engineering and hybridoma cells, can solve the problems of unreported neutralization activity, low neutralization titer, etc., to achieve antibody neutralization titer not reduced, high neutralization Good activity and stability

Active Publication Date: 2013-08-07
JIANGSU ACAD OF AGRI SCI
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AI Technical Summary

Problems solved by technology

At present, the hybridoma cell lines that secrete IBDV monoclonal antibodies have been reported. The monoclonal antibodies secreted by some cell lines have neutralizing activity against IBDV, but the neutralizing potency is low, and the neutralizing activity against different IBDV strains has not been seen. to report

Method used

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Experimental program
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Embodiment Construction

[0023] (1) Establishment of monoclonal antibody hybridoma cell lines

[0024] 1. Preparation of IBDV Antigen Isolation of virulent IBDV strains currently circulating from the bursa tissue of chickens with recent IBD vaccine immunization failure (Wang Yongshan et al., Molecular characteristics of the VP2 gene of infectious bursal disease virus that recently caused immune failure, Chinese Veterinary Science, 2008, 38(12):919-925), infected 4-week-old SPF chickens, collected the bursa tissues of dead chickens, made tissue homogenate, freeze-thawed three times, centrifuged at 9000rpm for 30min, took the supernatant, and used PEG precipitation and The discontinuous sucrose density gradient method was used for purification, the purified virus antigen was detected by sandwich ELISA, the virus concentration was measured by ultraviolet spectrophotometer, and it was stored at -70°C for future use.

[0025]

[0026] 2. Animal Immunization Purified IBDV was used as the immunogen to...

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Abstract

The invention relates to a hybridoma cell strain secreting a high-neutralization activity infectious bursal disease virus (IBDV) monoclonal antibody, and belongs to the field of biotechnology. The hybridoma cell strain C128 is screened from an established bank of hybridoma cells secreting the IBDV monoclonal antibody, and a mice ascites monoclonal antibody prepared by the hybridoma cell strain has IBDV neutralization titer of 1010. The high-neutralization activity IBDV monoclonal antibody secreted by the hybridoma cell strain C128 has high neutralization titer and a wide IBDV strain resistance range, can be used for clinical treatment on IBD-infected animals, and has obvious curative effects, good safety and no adverse side reaction. A monoclonal antibody preparation prepared by the hybridoma cell strain C128 keeps neutralization titer after being preserved for 2 years and has good stability.

Description

[0001] technical field [0002] The invention relates to a hybridoma cell secreting high neutralizing active infectious bursal virus (IBDV) monoclonal antibody, belongs to the field of biotechnology, and relates to antibody engineering technology. Specifically, the monoclonal antibody hybridoma C128 strain has excellent production performance, and the secreted monoclonal antibody not only has high neutralizing titer but also has high neutralizing activity against IBDV strains from different animal sources, and can be used for infectious bursae Clinical treatment of animals with IBD. Background technique [0003] Infectious Bursal Disease (IBD) is caused by Infectious Bursal Disease Virus (IBDV). characteristic infectious disease. The disease not only causes the death of sick animals, but also leads to immunosuppression of the body, which reduces the immune defense ability of the body and the failure of vaccination. It is one of the main infectious diseases that cause serio...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/20C07K16/10A61K39/42A61P31/14C12R1/91
Inventor 王永山吴晓悠诸玉梅夏兴霞毕振威潘群兴王晓丽欧阳伟董晨红
Owner JIANGSU ACAD OF AGRI SCI
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