High-density fermentation method of recombinant human thymalfasin
A new technology of high-density fermentation and thymus method, which is applied in the new high-density and high-expression fermentation field of recombinant human thymus method, can solve the problem that the yield cannot meet the scale requirements, fermentation scale, cell concentration and fusion protein expression Problems such as not being able to present simultaneously at a high level, low cell concentration and fusion protein expression, to achieve the effect of strong practicability and efficiency
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Embodiment 1
[0034] 1.30L fermenter pilot high-density fermentation:
[0035] 1) Working seed culture
[0036]Take a strain from the working seed bank at -80°C. After it is completely melted, take 100 μL of the bacterial solution and inoculate it in the primary seed medium (50 mL in a 250 mL Erlenmeyer bottle, containing 50 μg / mL kanamycin), and place in a shaker. The bed was cultivated, the temperature was controlled at 37°C, the rotating speed was 230rpm, and cultured for 8h to obtain the primary seed solution; the primary seed solution was inoculated into the secondary seed medium (800mL in 2L Erlenmeyer flasks containing 50μg / mL kanamycin), placed on a shaker for cultivation, controlled temperature at 37°C, rotation speed at 230rpm, and cultivated for 15-17h to obtain fermentation working seed liquid.
[0037] 2) Inoculate in a 30L fermenter for fermentation
[0038] Inoculate the prepared fermented working seed solution with 10% inoculum in a 30L fermenter containing 14L fermentati...
Embodiment 2
[0070] 1. 100L fermenter pilot high-density fermentation:
[0071] 1) Working seed culture
[0072] Take a strain from the working seed bank at -80°C. After it is completely melted, take 200 μL of the bacterial solution and inoculate it into the primary seed medium (100 mL in a 500 mL Erlenmeyer bottle, containing 50 μg / mL kanamycin), and place in a shaker. The bed is cultivated, the temperature is controlled at 37°C, the rotating speed is 230rpm, and it is cultivated for 8 hours to obtain the first-level seed solution; the first-level seed solution is inoculated on the second-level seed medium with an inoculation amount of 3% (4 2L Erlenmeyer flasks are respectively filled with 800mL, containing 50 μg / mL kanamycin), placed in a shaker for cultivation, controlled temperature at 37° C., rotation speed at 230 rpm, and cultivated for 15-17 hours to obtain fermentation working seed liquid.
[0073] 2) Inoculate in a 100L fermenter for fermentation
[0074] Inoculate the prepared...
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