A kind of preparation method of Haemophilus influenzae type b polysaccharide conjugate vaccine
A Haemophilus influenzae and conjugate vaccine technology, which is applied in the field of preparation of Haemophilus influenzae type B polysaccharide conjugate vaccine, can solve the problems of unstable quality, complicated process and high cost
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Embodiment 1
[0093] vaccine preparation
[0094] 1. Culture of Haemophilus influenzae type B;
[0095] The formula of the culture medium is that each liter of culture fluid contains the following components:
[0096]
[0097] The culture method is: shaking culture at 37 degrees Celsius and 150 rpm, after entering the logarithmic growth phase, adjust the pH to 7.4 with NaOH every 2 hours, co-cultivate for 24 hours, and sterilize with 0.6% formaldehyde after the culture.
[0098] 2. Extraction of polysaccharide extract of Haemophilus influenzae type B;
[0099] A, Extraction of PRP crude sugar: 1) Haemophilus influenzae type B culture solution 8000rpm10min to remove bacterial sediment, collect the supernatant; 2) pass the supernatant through a 0.22um hollow fiber, and harvest the filtrate; 3) Use the hollow fiber filtrate 50KDa membrane bag ultrafiltration concentration; 4) The membrane bag concentrated solution is subjected to graded alcohol precipitation: a. 25% low concentration alco...
Embodiment 2
[0104] Same as the method of Example 1, but in the cultivation of Haemophilus influenzae type B;
[0105] The formula of the culture medium is that each liter of culture fluid contains the following components:
[0106]
[0107] The culture method is: shake culture at 35°C, adjust the pH to 7 with NaOH every 1 hour after entering the logarithmic growth phase, co-cultivate for 24 hours, and sterilize after the culture is completed.
Embodiment 3
[0109] Same as the method of Example 1, but in the cultivation of Haemophilus influenzae type B;
[0110] The formula of the culture medium is that each liter of culture fluid contains the following components:
[0111]
[0112]
[0113] The culture method is: shake culture at 40°C, adjust the pH to 8 with NaOH every 3 hours after entering the logarithmic growth phase, co-cultivate for 24 hours, and sterilize after the culture is completed.
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