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FPV (feline parvovirus)-resistant cat-derived genetic engineering antibody

A technology of genetically engineered antibody and feline parvovirus, which is applied in the field of antibody engineering, can solve problems such as allergic reactions, reduced therapeutic effect, and inability to continuously inject, and achieve the effects of reducing immunogenicity and neutralizing the activity of FPV virus

Inactive Publication Date: 2019-09-27
CHANGCHUN SR BIOLOGICAL TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Anti-mouse antibodies are easily produced when mouse monoclonal antibodies are injected into animals of other species, which greatly reduces the therapeutic effect and limits clinical application
Moreover, most of the commercially available anti-FPV antibodies are immune sera of heterogeneous animals. Although they can protect young animals and adult animals from infection, they cannot be injected continuously, otherwise severe allergic reactions will be caused, and in severe cases, the animals may die

Method used

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  • FPV (feline parvovirus)-resistant cat-derived genetic engineering antibody
  • FPV (feline parvovirus)-resistant cat-derived genetic engineering antibody
  • FPV (feline parvovirus)-resistant cat-derived genetic engineering antibody

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0020] Embodiment 1 Feline transformation of murine CPV monoclonal antibody

[0021] (1) Construction of feline CPV antibody

[0022] Using the method of the patent application description example of the publication number CN109627331A, the mouse CPV monoclonal antibody was obtained, and the nucleotide sequence of its heavy chain variable region was sequenced as shown in SEQ ID NO.1, and the nucleotide sequence of its light chain variable region was The nucleotide sequence is shown in SEQ ID NO.2, connected to the cloning vector, the clones were picked for sequencing, and the sequencing results were analyzed online using IMGT to obtain the heavy chain amino acid sequence of the antibody sequence as shown in SEQ ID NO.3, and the light chain The amino acid sequence is shown in SEQ ID NO.4.

[0023] The heavy chain variable region gene sequence of the obtained mouse CPV monoclonal antibody was assembled with the felinized antibody heavy chain constant region gene sequence (as sh...

Embodiment 2

[0024] Example 2 Expression and Activity Research of Feline-derived CPV Genetic Engineering Antibody

[0025] (1) Eukaryotic expression of feline-derived CPV genetic engineering antibody

[0026] Take 1×10 6 cells / well cell density HEK-293 adherent cells were seeded in a six-well plate and grown for 12 hours before transient transfection experiments. The light chain:heavy chain=1:1 mixed with PEI (1 μg / μl) was transfected at a ratio of 1:3 and expressed in HEK-293 cells. The supernatant was collected 72 hours after transfection for activity identification.

[0027] (2) Hemagglutination inhibition method was used to measure the resistance effect of the feline-derived CPV genetically engineered antibody in the supernatant on the combination of feline parvovirus and red blood cells

[0028] Prepare the diluted FPV as eight units of virus (that is, the FPV virus solution with three hemagglutination units). Add 15 mm PBS with pH 6.5 to the 96-well V-type hemagglutination plate, 2...

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Abstract

The invention discloses an FPV (feline parvovirus)-resistant cat-derived genetic engineering antibody, and belongs to the technical field of antibody engineering. The antibody comprises a mouse-derived variable region and a cat-derived constant region, the nucleotide sequence of a heavy chain variable region of the antibody is represented as SEQ ID NO.1, the gene sequence of a heavy chain constant region of the antibody is represented as SEQ ID NO.5, the nucleotide sequence of a light chain variable region of the antibody is represented as SEQ ID NO.2, and the gene sequence of a light chain constant region of the antibody is represented as SEQ ID NO.6. The variable region sequence of a monoclonal antibody of a canine parvovirus is assembled with the cat-derived constant region, and the FPV-resistant cat-derived genetic engineering antibody is obtained, shows good FPV neutralization activity, has an effect of inhibiting red cell agglutination of the FPV, can be applied to the fields of cat-derived research for a canine parvovirus monoclonal antibody and the like and has great significance for promoting development of the cat-derived monoclonal antibody drugs.

Description

technical field [0001] The invention relates to the technical field of antibody engineering, in particular to a feline-derived genetic engineering antibody against feline parvovirus. Background technique [0002] Feline parvovirus (Feline parvovirus, FPV), which belongs to the family Parvoviridae and the genus Parvovirus, is the causative agent of panleukopenia in cats and felines, and has the characteristics of susceptibility and high mortality to young cats. With the gradual increase of pet owners, the number of pet cats is increasing year by year, and the prevention and treatment of cat parvovirus has attracted much attention. [0003] At present, the main clinical treatment method for feline parvovirus is to inject a large number of antiviral monoclonal antibodies to infected animals. Because mouse monoclonal antibody is easy to produce anti-mouse antibody when injected into animals of other species, the therapeutic effect is greatly reduced, which limits clinical appli...

Claims

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Application Information

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IPC IPC(8): C07K16/08A61K39/42A61P31/20
CPCA61K2039/505A61P31/20C07K16/081C07K2317/52C07K2317/56
Inventor 石晶李雪殷玉和李希辰赵健付玉张馨月
Owner CHANGCHUN SR BIOLOGICAL TECH
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