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Haemophilus influenzac type B polysaccharide conjugate vaccine preparation method

A Haemophilus influenzae-conjugate vaccine technology, which is applied in the field of preparation of Haemophilus influenzae type B polysaccharide conjugate vaccine, can solve the problems of high cost, complicated process and unstable quality

Active Publication Date: 2014-03-12
TASLY BIOPHARMACEUTICALS CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The existing Hib conjugate vaccine production technology mainly includes the selection and cultivation of strains, the extraction, separation and purification of capsular polysaccharide, the cultivation, separation and purification of carrier protein, the combination of capsular polysaccharide and carrier protein, and the separation and purification of vaccine. , where the key steps have defects such as complex process, high cost, and unstable quality

Method used

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  • Haemophilus influenzac type B polysaccharide conjugate vaccine preparation method
  • Haemophilus influenzac type B polysaccharide conjugate vaccine preparation method
  • Haemophilus influenzac type B polysaccharide conjugate vaccine preparation method

Examples

Experimental program
Comparison scheme
Effect test

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 at 8000rpm for 10min to remove the bacterial precipitate, and collect the supernatant; 2) Pass the supernatant through a 0.22um hollow fiber, and harvest the filtrate; 3) Hollow fiber filtrate Concentrate with 50KDa membrane bag ultrafiltration; 4) Carry out graded alcohol precipitation of the membrane bag concentrated solution: a. 25% low concent...

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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Abstract

The present invention discloses a Haemophilus influenzac type B polysaccharide conjugate vaccine preparation method, which comprises: A, preparing a Haemophilus influenzac type B polysaccharide extract (PRP), adopting ultrapure water or water for injection to dissolve the PRP polysaccharide at a room temperature until achieving a concentration of 20 mg / ml, adding CDAP according to a mass ratio of PRP to CDAP of 1, maintaining for 1.5 min, adjusting the pH value to 9.5 with 0.3 M NaOH, maintaining for 3 min, adding a solid ADH according to a mass ratio of ADH to the PRP polysaccharide of 4:1, adjusting the pH value to 9.5 with 0.3 M NaOH, maintaining for 1 h, and carrying out dialysis at a temperature of 2-8 DEG C with 0.2 M NaCl; and B, mixing the obtained PRP-AH and a 20 mg / ml TT solution according to a mass ratio of 1:2, adjusting the pH value to 5.0 with 0.1 M HCL, adding a solid EDAC according to a mass ratio of EDAC to PRP of 10:1, maintaining the pH value of 5.0 for 60 min at a room temperature, adjusting the pH value to 7.5 with 0.3 M NaOH, and carrying out a reaction for 30 min.

Description

technical field [0001] The invention relates to a preparation method of a vaccine, in particular to a preparation method of a Haemophilus influenzae type B polysaccharide-conjugated vaccine. Background technique [0002] Haemophilus influenzae type b (Hib) is a highly invasive type of Haemophilus influenzae. Almost 90% of the serious infections caused by Haemophilus influenzae are caused by type B, mainly causing meningitis, Pneumonia, pericarditis, arthritis, bacteremia, epiglottitis and other diseases, the disease makes 15% to 35% of survivors suffer from permanent disability, typical symptoms include mental retardation or deafness. Meningitis and pneumonia caused by Haemophilus influenzae type b are the leading causes of infant mortality. [0003] Years of epidemiological studies have shown that the use of Hib vaccine is an effective measure to control Hib invasive disease. Therefore, it is necessary and important to develop a Hib vaccine with good effect and low cost. ...

Claims

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Application Information

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IPC IPC(8): A61K39/385A61P31/04C08B37/00C12N1/20C12P21/02C07K14/34C07K1/36C07K1/34C07K1/30C07K1/20C07K1/18A61K39/102C12R1/16
Inventor 李军强孙倩赵秋敏李亚冰刘贺军金永杰
Owner TASLY BIOPHARMACEUTICALS CO LTD
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