Optimization Of The Expanded Culture Technology Of Porcine Interferon-γ And Porcine Interferon-α

In order to meet the requirement of green-farming.Our lab has used the insect baculovirus expression system obtained in such aspects as the structure and function was similar to natural porcine IFN-αIFN-γand determine the IFN-αIFN-γantiviral activities.On this basis.further study on the use of baculovirus expression vector of interferon production process,which realized the large-scale production of interferon in the laboratory as well as the exploration of interferon product storage methods and the method of baculovirus inactivation.It provides the technical support for the actual large-scale production.In this study.the interferon expression was expanded from the original 5 mL culture system to the 200 mL system in the laboratory and the optimal growth conditions for the expanded system of SF9 cells were explored.We found that after 1 day of culture in the shake flask.the cell density reached 1.3×10~6 cells/mL in the logarithmic growth phase.In order to determine the best amount of poison and the best time of poison collection.we added 200μL 400μL 2 mL 4 mL 6 mL 8 mL 12 mL of interferon seed poison into the shake flask,and collected the interference every day after receiving the poison for three days.Sample for antiviral activity test.We found that the amount of IFN-γwas 4mL.which is 0.00053 MOI and the antiviral activity of interferon was the best three days after inoculation.IFN-αhas the best antiviral effect in the dose of 200μL which is 0.00017 MOI and the interferon collected 3days after ingestion has the best antiviral effect.In order to ensure that the expressed interferon has high biological safety.the inactivation conditions of the recombinant baculovirus are optimized.We found that usingβ-propiolactone(BPL)at a dilution of 1:2000 and inactivating at 4℃ for 24 h can completely inactivate the baculovirus in IFN-γ.and then BPL is degraded at 37℃.The BPL can be completely degraded in 12 h without affecting the cells.We explored the storage time and storage conditions of interferon.Store the produced interferon at 4℃-20℃-80℃ We found that the best storage method is-20℃ for 2 months.First of all.this study completed the process optimization of IFN-αand IFN-γ200 mL culture system.Next we explored the process of inactivating baculovirus was explored.and the storage conditions and storage methods were.Provided important technical support for the actual large-scale production and application of interferon clinical.