Method for producing porcine reproductive and respiratory syndrome virus (PRRSV) antigen through whole suspension cell culture
A technology for respiratory syndrome and cell culture, applied in the field of full suspension cell culture to produce porcine reproductive and respiratory syndrome virus antigens, to achieve the effects of lower production costs, easy quality control, simple operation and easy expansion
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Embodiment 1
[0029] ——Production of porcine reproductive and respiratory syndrome virus antigen by whole suspension cell culture
[0030] 1. Preparation of porcine reproductive and respiratory syndrome virus antigen
[0031] (1) Cultivation and passage of seed cells
[0032] Seed cells MARC-145 were cultured and grown in shake flasks, samples were taken for cell counting, and the cell density reached 2.7×10 6 cells / ml, after adding serum-free cell culture medium MARC401, according to 0.5×10 6 The density of individual / ml is added in the reactor, and subculture under the condition of 37 ℃, 120r / min rotating speed;
[0033]The culture volume of the seed cell MARC-145 in the reactor was 5L of the final culture solution volume of the reactor.
[0034] (2) Inoculation of seed cells
[0035] The cell density of the seeded cell MARC-145 reaches 3.1×10 6 cells / ml, when the average diameter of the cells is 16.99 μm, the PRRSV-N3544 (CGMCC No.10398) venom of 5% of the final culture medium volum...
Embodiment 2
[0050] ——Production of porcine reproductive and respiratory syndrome virus antigen by whole suspension cell culture
[0051] 1. Preparation of porcine reproductive and respiratory syndrome virus antigen
[0052] (1) Cultivation and passage of seed cells in shake flasks
[0053] Seed cells MARC-145 were cultured and grown in shake flasks, samples were taken for cell counting, and the cell density reached 3.3×10 6 cells / ml, after adding serum-free cell culture medium MARC401, according to 1.0×10 6 The density of each / ml is added in the reactor, and subculture under the condition of 37 ℃, 100r / min rotating speed;
[0054] The culture volume of the seed cell MARC-145 in the reactor was 5L of the final culture solution volume of the reactor.
[0055] (2) Inoculation of seed cells
[0056] The cell density of the seeded cell MARC-145 reaches 3.4×10 6 Individual / ml, when the average cell diameter is 17.21 μm, the PRRSV-N3544 (CGMCC No.10398) kind of venom of 4% of the final cult...
Embodiment 3
[0071] ——Production of porcine reproductive and respiratory syndrome virus antigen by whole suspension cell culture
[0072] 1. Preparation of porcine reproductive and respiratory syndrome virus antigen
[0073] (1) Cultivation and passage of seed cells in shake flasks
[0074] Seed cells MARC-145 were cultured and grown in shake flasks, samples were taken for cell counting, and the cell density reached 2.9×10 6 cells / ml, after adding serum-free cell culture medium MARC401, according to 1.0×10 6 The density of each / ml is added in the reactor, and subculture under the condition of 37 ℃, 80r / min rotating speed;
[0075] The culture volume of the seed cell MARC-145 in the reactor was 5L of the final culture solution volume of the reactor.
[0076] (2) Inoculation of seed cells
[0077] The cell density of the seeded cell MARC-145 reaches 2.6×10 6 Individual / ml, when the average cell diameter is 17.03 μm, the PRRSV-N3544 (CGMCC No.10398) kind of venom of 3% of the final cultu...
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