Human papilloma virus L1 protein fermentation method
A technology of human papillomavirus and fermentation method, applied in the direction of microorganism-based method, fermentation, virus, etc., can solve the problems of HPV vaccine L1 protein effect is not obvious, it is difficult to determine the protease related to L1 protein degradation, and achieve the reduction of L1 protein Effect of degradation and cost reduction
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Embodiment 1
[0018] (1) Strain culture: Take 2ml of KM71 / A11-20 glycerol preservation solution and put it into a 250ml shake flask containing 100ml of BMGY seed solution medium, and culture at 30°C and 220r / min for 12h. Then take 2ml of the above-mentioned cultured seed solution and transfer them to three 500ml shake flasks containing 200ml of BMGY secondary seed solution medium, and cultivate them at 30°C and 220r / min for 18h.
[0019] (2) Inoculation: Inject 20L of L-GJY fermentation medium into a 50L fermenter, and then insert the above-mentioned cultured secondary seed liquid at an inoculation amount of 2.5% by volume.
[0020] (3) Fermentation: ①Basic growth stage: the initial rotation speed is 100r / min, the temperature is controlled at 30°C, the ventilation rate is 1vvm, the dissolved oxygen is controlled above 50%, and the pH does not need to be controlled. ②Glycerol supplementation stage: When the dissolved oxygen rebounds to 80%, add glycerin solution (50%, w / v, containing 12ml / L ...
Embodiment 2
[0023] (1) Strain culture: Take 2ml of KM71 / A11-20 glycerol preservation solution and put it into a 250ml shake flask containing 100ml of BMGY seed solution medium, and culture at 30°C and 220r / min for 12h. Then take 2ml of the above-mentioned cultured seed solution and transfer them to three 500ml shake flasks containing 200ml of BMGY secondary seed solution medium, and cultivate them at 30°C and 220r / min for 18h.
[0024] (2) Inoculation: Inject 20L of L-GJY fermentation medium into a 50L fermenter, and then insert the above-mentioned cultured secondary seed liquid at an inoculation amount of 2.5% by volume.
[0025] (3) Fermentation: ①Basic growth stage: the initial rotation speed is 100r / min, the temperature is controlled at 30°C, the ventilation rate is 1vvm, the dissolved oxygen is controlled above 50%, and the pH does not need to be controlled. ②Glycerol supplementation stage: When the dissolved oxygen rebounds to 80%, add glycerin solution (50%, w / v, containing 12ml / L ...
Embodiment 3
[0028] (1) Strain culture: Take 2ml of KM71 / A11-20 glycerol preservation solution and put it into a 250ml shake flask containing 100ml of BMGY seed solution medium, and culture at 30°C and 220r / min for 12h. Then take 2ml of the above-mentioned cultured seed solution and transfer them to three 500ml shake flasks containing 200ml of BMGY secondary seed solution medium, and cultivate them at 30°C and 220r / min for 18h.
[0029] (2) Inoculation: Inject 20LL-GJY fermentation medium into a 50L fermenter, and then insert the above-mentioned cultured secondary seed liquid at an inoculation amount of 2.5% by volume.
[0030](3) Fermentation: ①Basic growth stage: the initial rotation speed is 100r / min, the temperature is controlled at 30°C, the ventilation rate is 1vvm, the dissolved oxygen is controlled above 50%, and the pH does not need to be controlled. ②Glycerol supplementation stage: When the dissolved oxygen rebounds to 80%, add glycerin solution (50%, w / v, containing 12ml / L PTM1 ...
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