Canine parvovirus resistant genetic engineering antibody and application thereof
A technology of genetically engineered antibodies and canine parvovirus, applied in genetic engineering, antiviral agents, antiviral immunoglobulins, etc., can solve problems such as reducing curative effect, and achieve the effect of inhibiting agglutination and good neutralizing activity
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Embodiment 1
[0035] Canineization of murine CPV monoclonal antibody
[0036] (1) Construction of canine CPV genetic engineering antibody
[0037] 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.
[0038] The heavy chain variable region gene sequence of the obtained mouse CPV monoclonal antibody was assembled with the canine antibody heavy chain constant region gene sequence (as shown ...
Embodiment 2
[0040] Expression and activity of canine CPV genetically engineered antibody
[0041] (1) Eukaryotic expression of canine CPV genetic engineering antibody
[0042] Take 1×10 6 Cells / well cell density HEK-293 adherent cells were seeded in six-well plates and grown for 12 hours before transient transfection experiments. According to light chain:heavy chain = 1:1 mixed with PEI (1 μg / μL) at a ratio of 1:3, it was transfected into HEK-293 cells for expression, and the supernatant was collected 72 hours after transfection for activity identification.
[0043] (2) Hemagglutination inhibition method was used to measure the resistance effect of the canine-derived CPV genetically engineered antibody in the supernatant on the binding of canine parvovirus and porcine red blood cells
[0044] Add 25 μL / well of PBS (0.15M pH6.5) to the 96-well V-type hemagglutination plate, add 25 μL of canine CPV genetic engineering antibody to the first column, and do 3 replicates for each antibody, an...
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