Method for preparing pseudorabies virus
A technology of pseudorabies virus and pseudorabies virus, applied in biochemical equipment and methods, viruses/bacteriophages, microorganisms, etc., can solve the problems of low virus yield, large batch-to-batch difference, low virus titer, etc., and increase vaccine production , Improved safety, and genetically stable effects
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Embodiment 1
[0056] Embodiment 1 Utilizes Vero cells to continuously produce pseudorabies virus in a tidal bioreactor
[0057] Bioreactor: TideCell bioreactor, purchased from Saiyu Cell Technology Co., Ltd.
[0058] Carrier: BioNOC II carrier, purchased from Saiyu Cell Technology Co., Ltd.
[0059] CVD Nucleus Counting Kit: purchased from Saiyu Cell Technology Co., Ltd., Cat. No. AB0100, Lot No. 3007.
[0060] Glucose analyzer: purchased from Saiyu Cell Technology Co., Ltd.
[0061] Pseudorabies virus: Bartha-K61 strain (purchased from China Veterinary Drug Control Institute).
[0062] Medium: DMEM medium was purchased from GIBCO Company (product number: 12800-082, lot number: 1309922).
[0063]Trypsin: purchased from BD-Difco (product number: 215250, lot number: 5213753).
[0064] Fetal bovine serum: purchased from PAA company (product number: A 15-151, batch number: A64905-0972)
[0065] Glucose: purchased from SIGMA Company (article number: 7021, batch number: MFCD00063774).
[00...
Embodiment 2
[0073] Example 2 BHK-21 cells continuously produce pseudorabies virus in a tidal bioreactor
[0074] Cell culture medium: MEM medium was purchased from GIBCO Company (product number: 61100-087, batch number: 678277), and prepared into cell culture medium as described in the product manual.
[0075] Bovine serum: purchased from Hyclone Company (article number: SV30087.02, batch number: NSK0069)
[0076] Pseudorabies virus: Fa gene deletion strain (TK - / gE - / gI - ) (see Chinese patent CN101186902A for this strain, which is deposited in the China Center for Type Culture Collection (CCTCC) with the accession number V200002).
[0077] Other materials and reagents are the same as in Example 1.
[0078] The BHK-21 cells used in this example are deposited in the American Type Culture Collection (ATCC) with the accession number CCL-10. For details, see the ATCC web page http: / / www.atcc.org / ATCCAdvancedCatalogSearch / ProductDetails / tabid / 452 / Default. aspx? ATCCNum=CCL-10&Template...
Embodiment 3
[0080] Example 3 MDBK cells continuously produce pseudorabies virus in a tidal bioreactor
[0081]Cell culture medium: MEM medium was purchased from GIBCO Company (product number: 61100-087, batch number: 678277), and prepared into cell culture medium as described in the product manual.
[0082] Neonatal bovine serum (manufactured by GIBCO, product number 16010-14, batch number 693745)
[0083] Other material and reagent are with embodiment 1.
[0084] In this example, the MDBK cell (NBL-1) was deposited in the American Type Culture Collection (ATCC) with the accession number CCL-22. For details, please refer to the ATCC webpage. http: / / www.atcc.org / ATCCAdvancedCatalogSearch / ProductDetails / tabid / 452 / Default.aspx? ATCCNum=CCL-22&Template=cellBiology. In this embodiment, a tidal bioreactor with a carrier tank of 10 L is adopted. The steps, methods and parameters adopted are basically the same as those in Example 1, and the cell growth solution and cell maintenance solution us...
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