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Hybridoma Cell Strain a135 Efficiently Secreting Anti-Canine Parvovirus Monoclonal Antibody

A technology of canine parvovirus and monoclonal antibody, which is applied in antiviral agents, antiviral immunoglobulins, antibodies, etc., can solve the problems of infection of sick dogs, neutralization potency (low antiviral activity, high production cost, etc. Achieve a wide range of effects with excellent biological activity

Inactive Publication Date: 2017-12-26
JIANGSU ACAD OF AGRI SCI
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Problems solved by technology

At present, the anti-CPV immune serum prepared from native animals or heterologous animals in the market has unstable quality, difficult quality control, high production costs, and there is a huge risk of exogenous virus introduction and infection of sick dogs, resulting in serious Biosafety issues; the application of monoclonal antibody therapy can reduce the above-mentioned risks and improve the curative effect, but the existing anti-CPV monoclonal antibodies not only have low neutralization titer (antiviral activity), but also have low neutralization potency to different CPV subtypes. Potency varies widely, no neutralizing activity against some CPV subtype strains

Method used

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Embodiment Construction

[0018] (1) Establishment of monoclonal antibody hybridoma cell lines

[0019] 1. Preparation of CPV antigen The virulent canine parvovirus CPV-JS12 strain isolated from CPVD immunized dogs (Li Yuehua, et al. Molecular characteristics of VP2 gene of canine parvovirus JS12 strain and its expression in Escherichia coli. Chinese Veterinary Science, 2013, 43(10): 991-998) were inoculated in FK81 cells (purchased from Nanjing Tianbang Biotechnology Co., Ltd.), the diseased cell culture was frozen and thawed three times, and the cell debris was removed by centrifugation, and the supernatant was taken The solution was purified by PEG precipitation and discontinuous sucrose density gradient method, and the virus concentration was measured by ultraviolet spectrophotometer to prepare CPV immune antigen, which was stored at -20°C for future use.

[0020] Animal immunization Immunize female BALB / c mice aged 6-8 weeks (purchased from the Experimental Center of Comparative Medicine of Yangzh...

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Abstract

The invention relates to a hybridoma cell A135 strain highly secreting anti-canine parvovirus (CPV) monoclonal antibody, and belongs to the field of biotechnology. The present invention screens out a hybridoma cell A135 strain from the established hybridoma cell bank secreting anti-CPV monoclonal antibody, which has excellent biological properties, and the neutralizing effect on CPV by injecting the ascites induced in the peritoneal cavity of BALB / C mice The valency is as high as 1010, and the neutralization ability to CPV-2a, CPV-2b, CPV-2c(a), CPV-2c(b) and other CPV subtype strains, as well as fox and raccoon-derived CPV strains, is equivalent. The range of CPV strains is wide, and it is used for the clinical treatment of dogs with CPV infection, and the effective rate reaches 100%.

Description

technical field [0001] The invention relates to a hybridoma cell highly secreting anti-canine parvovirus (CPV) monoclonal antibody, belonging to the field of biotechnology. Background technique [0002] Canine parvovirus disease (CPVD) is a canine viral infectious disease caused by Canine parvovirus (CPV), characterized by hemorrhagic enteritis or non-suppurative myocarditis. The disease is highly contagious. The mortality rate is high, and dogs of all ages can be infected, and the mortality rate of puppies is as high as 70%, and the economic loss caused by the disease is huge. Although CPVD vaccination has been widely adopted for susceptible dogs, CPV is still one of the most serious infectious diseases that endanger the dog industry in my country because of its strong resistance to the outside world and easy mutation. [0003] CPV (also known as CPV-2) is a new type of parvovirus isolated from the stool of dogs with hemorrhagic enteritis by American scholars in 1977. Vir...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/20C07K16/08A61K39/395A61P31/20
Inventor 王永山夏兴霞毕振威李月华诸玉梅欧阳伟潘群兴王晓丽董晨红梅永杰王晶宇吴红玲
Owner JIANGSU ACAD OF AGRI SCI
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