Haemophilus influenzae type b (Hib) polysaccharide and refined tetanus toxoid coupling process

A technology of tetanus toxoid and polysaccharide, which is applied in the directions of inactive medical preparations, carrier-binding antigen/hapten components, antibacterial drugs, etc., can solve the problems of wasting time and solution, easily polluted, etc. Dosage, reduce the chance of pollution, improve work efficiency

Active Publication Date: 2013-01-09
CHENGDU OLYMVAX BIOPHARM
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Many pharmaceutical companies use traditional dialysis methods to remove hydrogen bromide, adipic hydrazide and carbodiimide, but dialysis usually ta

Method used

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  • Haemophilus influenzae type b (Hib) polysaccharide and refined tetanus toxoid coupling process
  • Haemophilus influenzae type b (Hib) polysaccharide and refined tetanus toxoid coupling process

Examples

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Embodiment 1

[0021] A Hib polysaccharide and refined tetanus toxoid coupling process, which includes two steps of Hib polysaccharide-AH derivative generation and Hib polysaccharide-TT conjugate preparation,

[0022] The generation of the Hib polysaccharide-AH derivative comprises the following substeps:

[0023] A1, dissolving the polysaccharide, weighing the Hib polysaccharide, adding sterilized water for injection, so that the final concentration is 1mg / ml;

[0024] A2, polysaccharide activation, according to the amount of 0.3 times Hib polysaccharide, add CNBr to activate, and keep the pH at 9.8 and react for 25 minutes;

[0025] A3, generate, add ADH 3 times the amount of Hib polysaccharide for connection, and keep the pH at 8.0 for 20-30 minutes;

[0026] A4. For purification, use a 30KD membrane bag, first dilute with 0.05mol / L NaCl at a volume ratio of 1:1, then ultrafilter to 1 / 2 volume, repeat dilution and ultrafiltration 13 times. After completion, filter with a 0.45 μm filter ...

Embodiment 2

[0035] A Hib polysaccharide and refined tetanus toxoid coupling process, which includes two steps of Hib polysaccharide-AH derivative generation and Hib polysaccharide-TT conjugate preparation,

[0036] The generation of the Hib polysaccharide-AH derivative comprises the following substeps:

[0037] A1, dissolving the polysaccharide, weighing the Hib polysaccharide, adding sterilized water for injection, so that the final concentration is 2 mg / ml;

[0038] A2, polysaccharide activation, add CNBr according to the amount of 0.2 times Hib polysaccharide to activate, and keep the pH at 9.5 and react for 30 minutes;

[0039] A3, generate, add ADH 5 times the amount of Hib polysaccharide for connection, and keep the pH at 8.5 for 20 minutes;

[0040] A4. For purification, use a 50KD membrane bag, first dilute with 0.02mol / L NaCl at a volume ratio of 1:1, then ultrafilter to 1 / 2 volume, repeat dilution and ultrafiltration 15 times. After completion, filter with a 0.45 μm filter mem...

Embodiment 3

[0049] A Hib polysaccharide and refined tetanus toxoid coupling process, which includes two steps of Hib polysaccharide-AH derivative generation and Hib polysaccharide-TT conjugate preparation,

[0050] The generation of the Hib polysaccharide-AH derivative comprises the following substeps:

[0051] A1, dissolving the polysaccharide, weighing the Hib polysaccharide, adding sterilized water for injection, so that the final concentration is 1mg / ml;

[0052] A2, polysaccharide activation, add CNBr to activate according to the amount of 0.6 times Hib polysaccharide, and keep the pH at 10.5 for 20 minutes;

[0053] A3, generate, add ADH 2 times the amount of Hib polysaccharide for connection, and keep the pH at 7.5 for 30 minutes;

[0054] A4. For purification, use a 40KD membrane bag, first dilute with 0.03mol / L NaCl at a volume ratio of 1:1, then ultrafilter to 1 / 2 volume, repeat dilution and ultrafiltration 10 times. After completion, filter with a 0.45 μm filter membrane to o...

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Abstract

The invention discloses a haemophilus influenzae type b (Hib) polysaccharide and refined tetanus toxoid coupling process which includes steps of Hib polysaccharide-AH derivant generation and Hib polysaccharide-tetanus toxoid (TT) combination preparation. The Hib polysaccharide-AH derivant generation includes steps of A1, polysaccharide dissolving, A2, polysaccharide activation, A3, antidiuretic hormone addition for connection and A4, CNBr and antidiuretic hormone (ADH) removing. The Hib polysaccharide-TT combination preparation includes the following steps: B1, adding TT solution, B2, adding carbodiimide (EDAC), B3, removing the EDAC to obtain polysaccharide-TT carbodiimide; and B4, separating and pourifying the polysaccharide-TT carbodiimide, eluting, degerming and filtering to obtain the polysaccharide-TT combination. The Hib polysaccharide and refined tetanus toxoid coupling process has the advantages of improving working efficiency, saving time and labor and simultaneously saving using amount of NaCl solution and reduces the contaminated rate of samples compared with dialysis adopted by the traditional process.

Description

technical field [0001] The invention relates to a coupling technology of Hib polysaccharide and refined tetanus toxoid. Background technique [0002] Haemophilus influenzae type b Hib (Hamephilus influenzae type b, referred to as Hib) polysaccharide vaccine has produced a good immune effect on older children since it was launched in the United States in 1985, but it is not effective in inducing infants under the age of 18 months. Bactericidal antibodies can not induce immune memory, and the actual immune efficacy is very low, which hinders the popularization and use of Hib polysaccharide vaccine. The first Hib polysaccharide vaccine was launched in the United States in April 1985. This vaccine is suitable for children aged 2 to 5. For children younger than 18 months, the effective rate of this vaccine using PRP is actually zero, which has become the biggest obstacle in the use of this vaccine. According to epidemiological surveys, children under the age of 6 months need pr...

Claims

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Application Information

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IPC IPC(8): A61K39/385A61K47/48A61P31/04C08B37/00
Inventor 陈庚伍长华罗力心吴强关晓峰陈克平
Owner CHENGDU OLYMVAX BIOPHARM
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