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Hybrid tumor cell strain 1D7 capable of secreting high neutralizing activity canine distemper virus monoclonal antibody

A kind of hybridoma cell line, monoclonal antibody technology

Inactive Publication Date: 2013-06-12
JIANGSU ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there have been reports on hybridoma cell lines secreting CDV monoclonal antibodies. The monoclonal antibodies secreted by some cell lines have neutralizing activity against CDV, but the neutralizing potency is low, and the neutralizing activity against different CDV strains has not been reported so far. See the report

Method used

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Experimental program
Comparison scheme
Effect test

Embodiment Construction

[0020] (1) Establishment of monoclonal antibody hybridoma cell lines

[0021] 1. Preparation of CDV antigen The virulent strain of CDV NJ01 recently isolated from the tissues of immunized dogs (Bi Zhenwei et al., Analysis of nucleocapsid protein gene sequence of atypical canine distemper virus and its expression in Escherichia coli. China Prevention Veterinary Medicine Acta, 2011,33(8):625-629.) Inoculated in vero cells, 3d cell lesions (CPE), 5d after the virus collection. Lesioned cell cultures were repeatedly frozen and thawed three times, and cell debris were removed after centrifugation at 4000×g. After the virus supernatant was precipitated by 1M zinc acetate solution, the precipitate was resuspended with saturated EDTA solution, centrifuged at 49000×g for 60 min at 4°C, and the precipitate was fully suspended with NTE. Add 20%, 30%, 40%, 50% and 60% (mass / volume) density gradient sucrose solutions, and centrifuge at 61000×g for 90min. Aspirate the virus band, observe ...

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Abstract

The invention relates to a hybrid tumor cell strain capable of secreting high neutralizing activity CDV (canine distemper virus) monoclonal antibody. A hybrid tumor cell strain 1D7 is selected from a hybrid tumor cell bank containing 82 strains which secrete CDV monoclonal antibody, and is used for preparing mouse ascitic antibody which has neutralizing potency up to 106 to CDV. The hybrid tumor cell strain capable of secreting high neutralizing activity CDV monoclonal antibody is high in neutralizing potency and resistant to CDV strains in a wide range, and has evident treatment effect on CDattached dogs, minks or foxes in clinical treatment, and is safe without adverse side effects. The monoclonal antibody therapeutic agent prepared with the hybrid tumor cell strain has stable neutralizing potency after two years of storage, and is high in stability.

Description

technical field [0001] The invention relates to a hybridoma cell 1D7 strain secreting high neutralizing active canine distemper virus (CDV) monoclonal antibody, belongs to the field of biotechnology, and relates to antibody engineering technology. Specifically, it is a CDV monoclonal antibody with high neutralization activity, which has high neutralization activity against CDV strains from different animal sources, and can be used for clinical treatment of various canine distemper (CD)-affected animals such as dogs, minks and foxes. Background technique [0002] Canine distemper (CD) is a highly contagious disease caused by canine distemper virus (CDV) in dogs and various animals in the order Carnivora. The incidence rate can reach almost 100%, and the mortality rate is more than 80%. The morbidity and fatality rate of canine distemper in our country all have the rising trend, and the clinical manifestations are also different from the past. Typical CD is clinically charact...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/20C07K16/10A61K39/42A61P31/14C12R1/91
Inventor 王永山毕振威夏兴霞孙婧范红结赵艳兵温海秦海斌张汇东
Owner JIANGSU ACAD OF AGRI SCI
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