Preparation method and application of novel CpGISS vaccine adjuvant
A vaccine adjuvant and a new technology, applied in the field of preparation of new CpGISS vaccine adjuvants, can solve the problems that CpG preparations are difficult to enter cells, easily degraded by nucleases, and lose activity, so as to improve the survival rate and immune protection rate. The level of cellular immune response, the effect of rapid replication
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Embodiment 1
[0019] A preparation method of a novel CpGISS vaccine adjuvant, the preparation steps are:
[0020] 1) After the activation culture, primary shake flask culture and secondary shake flask culture are performed on the preserved Pichia pastoris, the strain is inoculated into a sterilized fermenter. The components of the fermentation medium and their parts by weight are: 50-60 parts of sucrose, 60-80 parts of yeast powder, 10-15 parts of peptone, 30-45 parts of urea, 6-9 parts of wort juice, KH 2 PO 4 8~12 copies, K 2 HPO 4 ·3H 2 O 5~7 parts, MgSO 4 ·7H 2 O 0.2~0.3 parts, ZnSO 4 ·7H 2 O 5~7.8 parts, MnSO 4 4H 2 O 24~35 parts, biotin 0.016~0.02 parts, riboflavin sodium phosphate 0.003~0.005 parts, eicosapentaenoic acid ethyl ester 0.02~0.06 parts and deionized water 1700~2000 parts. Adjust the rotation speed and air flow, and control the dissolved oxygen content in the culture medium to 30-38%. During the fermentation process, the pH of the culture medium is controlled b...
Embodiment 2
[0025] A kind of preparation method of novel CpGISS vaccine adjuvant, the most preferred steps are:
[0026] 1) After the activation culture, primary shake flask culture and secondary shake flask culture are performed on the preserved Pichia pastoris, the strain is inoculated into a sterilized fermenter. The components of the fermentation medium and the most preferred parts by weight are: 56 parts of sucrose, 72 parts of yeast powder, 13 parts of peptone, 40 parts of urea, 8 parts of wort, KH 2 PO 4 10 copies, K 2 HPO 4 ·3H 2 O 6 parts, MgSO 4 ·7H 2 O 0.25 parts, ZnSO 4 ·7H 2 O 7.2 parts, MnSO 4 4H 2 33 parts of O, 0.019 parts of biotin, 0.005 parts of riboflavin sodium phosphate, 0.04 parts of ethyl eicosapentaenoate and 1800 parts of deionized water. Adjust the rotation speed and air flow rate, and control the dissolved oxygen content in the culture medium to 36%. During the fermentation process, the pH of the culture medium is controlled to be 5.0 by adding ammon...
Embodiment 3
[0031] A preparation method of a novel CpGISS vaccine adjuvant, the preparation steps are:
[0032] 1) After the activation culture, primary shake flask culture and secondary shake flask culture are performed on the preserved Pichia pastoris, the strain is inoculated into a sterilized fermenter. The components of the fermentation medium and their parts by weight are: 54 parts of sucrose, 72 parts of yeast powder, 13 parts of peptone, 41 parts of urea, 7 parts of wort, KH 2 PO 4 10 copies, K 2 HPO 4 ·3H 2 O6 parts, MgSO 4 ·7H 2 O 0.2 parts, ZnSO 4 ·7H 2 O 6.5 parts, MnSO 4 4H 2 30 parts of O, 0.017 parts of biotin, 0.003 parts of riboflavin sodium phosphate, 0.04 parts of ethyl eicosapentaenoate and 1800 parts of deionized water. Adjust the rotating speed and air flow, and control the dissolved oxygen content in the culture medium to 35%. During the fermentation process, the pH of the culture medium is controlled to be 5.0 by adding ammonia water, and the culture tem...
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