A method for producing recombinant human interferon beta 1b protein by large-scale fermentation
A recombinant human interferon and fermenter technology, which is applied in the field of large-scale fermentation and production of recombinant human interferon β1b protein, can solve the problems of low yield of fermentation products and the inability to apply β interferon industrial production, etc., to increase fermentation yield, Improve the effect of cell harvest and high-efficiency expression
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Embodiment 1
[0018] 1. Prepare 300L 2YT medium in a 500L fermenter, sterilize at 121°C and cool down to 35°C.
[0019] 2. Inoculate 15L of mature recombinant Escherichia coli seed solution for cultivation. During the entire fermentation and cultivation stage, the temperature is maintained at 35±1°C, the rotation speed is 200r / min, the ventilation rate is 200L / min, and the pH is 5.0.
[0020] 3. During the fermentation and cultivation process, take samples regularly to measure the OD600 value of the cell density of the bacterial liquid; cultivate until the OD600 of the cell density reaches 2, and then add isopropylthiogalactopyranoside (IPTG) to a final concentration of 0.2mM for induction.
[0021] 4. Start feeding after induction, and the feeding medium is also 2YT medium. Feeding rate Feed-feeding was carried out at a rate of 5% of the original medium volume per hour. Continue to cultivate for 4 hours and the fermentation ends.
[0022] 5. Sampling was used to determine the expression ...
Embodiment 2
[0024] 1. Prepare 300L 2YT medium in a 500L fermenter, sterilize at 121°C and cool down to 37°C.
[0025] 2. Inoculate 30L of mature recombinant Escherichia coli seed solution for cultivation. During the entire fermentation and cultivation stage, the temperature is maintained at 37±1°C, the rotation speed is 300r / min, the ventilation rate is 300L / min, and the pH is 8.0.
[0026] 3. During the fermentation and cultivation process, take samples regularly to measure the OD600 value of the cell density of the bacterial liquid; cultivate until the OD600 of the cell density reaches 8, and then add isopropylthiogalactopyranoside (IPTG) to a final concentration of 0.8mM for induction.
[0027] 4. Start feeding after induction, and the feeding medium is also 2YT medium. Feeding rate Feed-feeding was carried out at a rate of 10% of the original medium volume per hour. Continue to cultivate for 2 hours and the fermentation ends.
[0028] 5. Sampling was used to determine the expression...
Embodiment 3
[0030] 1. Prepare 300L 2YT medium in a 500L fermenter, sterilize at 121°C and cool down to 40°C;
[0031] 2. Inoculate 45L of mature recombinant Escherichia coli seed solution for cultivation, and maintain the temperature at 40±1°C, rotating speed at 500r / min, ventilation rate at 500L / min, and pH at 7.0 during the entire fermentation period;
[0032] 3. During the fermentation and cultivation process, take samples regularly to measure the OD600 value of the cell density of the bacterial liquid; cultivate until the cell density OD600 reaches 5, and then add isopropylthiogalactopyranoside (IPTG) to a final concentration of 1.5mM for induction.
[0033] 4. Start feeding after induction, and the feeding medium is also 2YT medium. The feeding rate was fed by feeding at a rate of 1% of the original medium volume per hour. Continue to cultivate for 8 hours and the fermentation ends.
[0034] 5. Sampling was used to determine the expression level of the bacteria by SDS-PAGE electrop...
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