Polyvalent pneumococcal polysaccharide combination vaccine
A technology of Streptococcus pneumoniae and Streptococcus polysaccharide, applied in the field of immunology, can solve problems such as poor immunogenicity
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preparation example Construction
[0063] The invention also provides a preparation method of the Streptococcus pneumoniae polysaccharide-carrier protein conjugate. It includes steps:
[0064] A. Bacterial culture:
[0065] The culture techniques and conditions of Streptococcus pneumoniae can adopt conventional techniques and conditions in the art. For example, pick the strains from the production seed batch, inoculate in 5 ml of culture solution, expand to 500 ml of culture solution, and then expand to 5 liters of culture solution. The culture conditions are the conditions for culturing streptococcus in general.
[0066] B. Polysaccharide extraction:
[0067] When the bacteria were cultured to OD ≥ 1.5, the culture was stopped and sodium deoxycholate was added for sterilization.
[0068] Centrifuge to remove the cells and collect the supernatant.
[0069] Add ethanol to the supernatant to a final volume concentration of 70-90%, and centrifuge to collect precipitated crude polysaccharides. In other report...
Embodiment 1
[0091] Example 1 Selection of Streptococcus pneumoniae subtype
[0092]In my country, from May 1981 to July 1985, 712 strains of pneumococcus were isolated from different specimens of patients, and were typed according to the Danish method and standards recommended by WHO. They were divided into 42 types (groups), and no 40, 42 and 47 strains were found. Among them, type 5 was the most, followed by 6, 1, 19, 23, 2, 3 and 8, a total of 8 types (groups) with 453 strains, accounting for 63.6%. The main serotypes of different diseases are different. Pneumonia is mainly 1, 5, 14 and 6; meningitis is mainly 2, 5, 6, 1, 27 and 14; otitis media is 19, 6, 5, 23 , 3 and 14 types are common. The main epidemic strains of pneumococcus sometimes change. Therefore, bacterial type monitoring is a very important task. It is an important scientific basis for determining the combination of pneumococcal vaccine serotypes and actively preventing pneumococcal infection.
[0093] In practical appli...
Embodiment 2
[0095] Example 2 Preparation of Streptococcus pneumoniae capsular polysaccharide
[0096] A. Bacterial culture:
[0097] Streptococcus pneumoniae of subtypes 4, 6B, 9V, 14, 18C, 19F and 23F were selected respectively. Bacteria were selected from the production seed batch, inoculated in 5 milliliters of nutrient solution, expanded to 500 milliliters of nutrient solution, and then expanded to 5 liters of nutrient solution. The culture conditions are the conditions for culturing streptococcus in general.
[0098] B. Polysaccharide extraction:
[0099] (b1) When the bacteria are cultured to OD ≥ 1.5, stop the culture and add sodium deoxycholate to sterilize.
[0100] (b2) Centrifuge at 14000g, remove the bacteria, and collect the supernatant.
[0101] (b3) Add ethanol to the supernatant to concentrations of 70%, 80% and 90%, respectively.
[0102] (b4) centrifuging to remove impurities such as pigments and inorganic salts, and collecting precipitated crude polysaccharides.
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