Recombinant feline herpesvirus type 1 gb-gd protein and its application
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A feline herpes virus, gb-gd technology, applied in the direction of recombinant DNA technology, virus, application, etc., can solve the problem of not being able to prevent infection or disease development in cats with virus-carrying diseases, so as to improve stability and immunogenicity, reduce Production cost, effect of prolonging protein half-life
Active Publication Date: 2022-05-31
苏州沃美生物有限公司
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Summary
Abstract
Description
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Problems solved by technology
Existing FHV-1 vaccines confer reasonable protection against disease in naïve cats but do not prevent infection or disease development in infected cats
Method used
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Experimental program
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Effect test
Embodiment 1
[0065]
[0070]
[0072]
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[0078] Take 10 μl of the ligation product obtained in step (3) and add 100 μl of DH5α competent cells, mix well, and heat shock at 42° C. for 90
Embodiment 2
[0085]
[0086]
[0091]
[0093]
[0097]
[0098]
[0100] Take 10 μl of the ligation product obtained in step (3), add 100 μl of DH5α competent cells, mix well, and heat shock at 42° C. for 90
Embodiment 3
[0115] 2.3 When the well of a single cell in the 96-well plate grows up, discard the medium, wash once with PBS, 100 μl 0.25%
trypsin-EDTA, digest at room temperature for about 2 minutes, add 2mL CHO-WM medium (containing 10% FBS+50μM MSX) to stop the digestion
the reaction, and the cells were blown off with a pipette. The cells were transferred to a 12-well plate, and when the 12-well plate was full, the supernatant was removed, and the Elisa assay was performed.
Check whether the clones are positive, and the positive clones with high expression continue to be expanded and cultured and cryopreserved.
3. Cell shake flask fermentation
3.1 Configuration of subculture medium: use CHO-WM substratum to add 50 μM MSX as subculture medium, place at 37
°C water bath preheated to 37 °C.
3.2 From CO
2
Remove the cells from the shake flask and count them.
3.3 Dilute cells to 2.5-3.5×10
5
Cells / mL were seeded in 30 mL cultures based on a 125 mL shake flask. thin
Pla...
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Abstract
The invention discloses a recombinant feline herpesvirus type 1 gB-gD protein and application thereof. The recombinant feline herpesvirus type 1 gB-gD protein comprises: the first protein having the sequence shown in SEQ ID NO:2; and / or, the second protein having the sequence shown in SEQ ID NO:6. The vaccine comprises the recombinant feline herpesvirus type 1 gB-gD protein and a pharmaceutically acceptable carrier. The recombinant feline herpesvirus type 1 gB-gD protein provided by the invention has a long half-life, high molecular stability, can form dimers, especially heterodimers, and has strong immunogenicity. The preparation of large-scale serum-free suspension culture in bioreactor not only has high expression level and good protein immunogenicity, but also has the advantages of easy quality control, batch-to-batch stability, and low production cost.
Description
Recombinant feline herpesvirus type 1 gB-gD protein and its application technical field The present invention relates to a kind of genetic engineering vaccine, be specifically related to a kind of recombinant feline herpesvirus 1 type gB-gD protein and its The application, such as the application in the preparation of a feline herpesvirus type 1 recombinant subunit vaccine, belongs to the technical field of animal immune medicine. Background technique Feline Herpesvirus type 1 (Feline Herpesvirus type 1, FHV-1) is feline infectious rhinotracheitis The causative agent of Feline Infectious Rchiotracheitis (FIR) is the acute upper respiratory system of felines. One of the main pathogens of disease. FHV‑1 infects only felines, of which kittens are the most susceptible, with an incidence rate of 100%. After FHV‑1 infection, it mainly affects the upper respiratory tract and eyes of cats, and the sick cats will have elevated body temperature, depression, coughing, and snee...
Claims
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Application Information
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