Production method for efficiently secreting and expressing porcine epidemic diarrhea virus S1 protein
A technology for porcine epidemic diarrhea and secretion expression, applied in biochemical equipment and methods, viruses, viral peptides, etc., can solve problems such as losses and unsatisfactory effects in the pig industry, save labor and material costs, cost-effective, and improve stability. sexual effect
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Embodiment 1
[0029] Example 1 A production method for efficiently secreting and expressing porcine epidemic diarrhea virus S1 protein
[0030] The production method for the efficient secretion and expression of porcine epidemic diarrhea virus S1 protein comprises the following steps:
[0031] S1. Treatment of the cell bioreactor: Check whether the consumables are damaged, record the batch, inflate the torrent bag and the perfusion bag for leak detection overnight (note that the inflation flow rate is not too large, the filter membrane will be crushed and the bag will be discarded); DO electrode Power on the electrode for more than 6 hours; the next day, after the calibration electrode is sterilized, fill the tank and connect the pipeline. After the system is connected, check the air tightness, inject 2.5L of PBS buffer solution, and soak the paper carrier in the reactor overnight. On the third day, pour out PBS; add 2L DMEM complete medium to the above reactor, pretreat the polyfiber disc ...
Embodiment 2
[0041] Example 2 Comparison experiment of bioreactor expression and cell factory expression
[0042] Petri dish expression method:
[0043] 1) According to the culture conditions of the bioreactor in Example 1, inoculate in the 10-layer cell factory after cultivating enough cells in the cell culture dish, and use 300g / L glucose and 7.5%NaHCO 3 Adjust the reaction conditions in the petri dish to be consistent with the bioreactor;
[0044] 2) After the cells are full, replace the serum-free medium according to the ratio of the number of cells, and use 300g / L glucose and 7.5% NaHCO 3 Adjust the reaction conditions in the petri dish to be consistent with the bioreactor; collect 1 / 2-2 / 3 volume of serum-free medium expression supernatant every 4 days and supplement the corresponding volume of serum-free medium, which is the same as that of the bioreactor every 4 days. Batch comparison of protein expression.
[0045] As a result, only 2 batches of expression samples can be collect...
Embodiment 3
[0046] The impact of embodiment 3 different pH values on protein yield
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