Optimization method of Hib polysaccharide and protein binding process

An optimization method and polysaccharide technology, applied in the direction of carrier binding antigen/hapten components, pharmaceutical formulations, medical preparations containing active ingredients, etc., can solve the problem of low yield of qualified conjugates and achieve an increase in the yield of qualified conjugates , Conducive to large-scale production, the effect of improving the binding rate

Inactive Publication Date: 2012-08-15
CHENGDU OLYMVAX BIOPHARM
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  • Abstract
  • Description
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Problems solved by technology

However, the yield of qualified conjugates of this binding method is very low. At present, the yield of Hib capsular polysaccharide of this binding method i...

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  • Optimization method of Hib polysaccharide and protein binding process

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Embodiment 1

[0022] A kind of optimization method of Hib polysaccharide and protein binding technology, it comprises following polysaccharide derivation, polysaccharide-TT (tetanus toxoid protein) conjugate preparation two steps, wherein, described polysaccharide derivation comprises following sub-steps:

[0023] S11: Add water for injection to the Hib capsular polysaccharide, and dilute to a solution with a final concentration of 1 mg / ml;

[0024] S12: adding CNBr 0.1 times the mass of the polysaccharide for activation treatment;

[0025] S13: After the activation is completed, add SDH (sebacic acid dihydrazide) 5 times the mass of the polysaccharide to carry out the coupling reaction;

[0026] S14: Divide the reactants obtained from the coupling reaction into dialysis bags, and dialyze in 0.02mol / L NaCl solution for 3 times, each time for 8 hours, to remove residual cyanide, and then concentrate to the original volume by ultrafiltration , to obtain polysaccharide-SDH (sebacic acid dihyd...

Embodiment 2

[0039]A kind of optimization method of Hib polysaccharide and protein binding technology, it comprises following polysaccharide derivation, polysaccharide-TT (tetanus toxoid protein) conjugate preparation two steps, wherein, described polysaccharide derivation comprises following sub-steps:

[0040] S11: Add water for injection to the Hib capsular polysaccharide, and dilute to a solution with a final concentration of 1.5 mg / ml;

[0041] S12: adding CNBr 0.4 times the mass of the polysaccharide for activation treatment;

[0042] S13: After the activation is completed, add SDH (sebacic acid dihydrazide) 4 times the mass of the polysaccharide to carry out the coupling reaction;

[0043] S14: Divide the reactants obtained from the coupling reaction into dialysis bags, and dialyze twice in 0.08mol / L NaCl solution, each time for 12 hours, to remove residual cyanide, and then concentrate to the original volume by ultrafiltration , to obtain polysaccharide-SDH (sebacic acid dihydrazi...

Embodiment 3

[0050] A kind of optimization method of Hib polysaccharide and protein binding technology, it comprises following polysaccharide derivation, polysaccharide-TT (tetanus toxoid protein) conjugate preparation two steps, wherein, described polysaccharide derivation comprises following sub-steps:

[0051] S11: Add water for injection to the Hib capsular polysaccharide, and dilute to a solution with a final concentration of 2 mg / ml;

[0052] S12: adding CNBr 0.25 times the mass of the polysaccharide for activation treatment;

[0053] S13: After the activation is completed, add SDH (sebacic acid dihydrazide) 3 times the mass of the polysaccharide to carry out the coupling reaction;

[0054] S14: Divide the reactants obtained from the coupling reaction into dialysis bags, and dialyze in 0.05mol / L NaCl solution for 2.5 times, each time for 10 hours, to remove residual cyanide, and then concentrate to the original volume by ultrafiltration , to obtain polysaccharide-SDH (sebacic acid d...

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Abstract

The invention discloses an optimization method of Haemophilus Influenzae Serotype B polysaccharide and protein binding process, and the method comprises the following steps of: derivating polysaccharide, and preparing polysaccharide-TT (tetanus toxoid) conjugate. The invention provides a novel bridging substance combined by polysaccharide and vector protein, and the novel bridging substance can be used for more effectively performing binding reaction by changing a bridge molecule based on a conventional method, so that the binding rate of polysaccharide and vector protein is improved, the binding reaction time is shortened, the yield of qualified conjugate of the binding reaction is improved to a certain extent, the binding reaction has high repeatability, large-scale production is facilitated, and the production cost is reduced.

Description

technical field [0001] The invention relates to a method for optimizing the binding process of Hib polysaccharide and protein. Background technique [0002] Haemophilus influenzae (Haemophilus Influenzae, Hi) is still the main pathogen causing human invasive diseases so far, most of which are caused by Haemophilus Influenzae type b (Hib). Hib meningitis ranks first among bacterial meningitis, and has the characteristics of high morbidity, high fatality rate, and high disability rate. The age of onset is young, and the infection rate of infants is high. The age of onset is mainly concentrated under 5 years old, especially 2 years old. the following. According to the report of the World Health Organization in 2006, at least 3 million cases of severe diseases are caused every year in the world, and about 386,000 of them die, which has become a major global public health problem. In developing countries, Hib pneumonia plays an important role in children's infectious diseases, ...

Claims

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Application Information

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IPC IPC(8): C08B37/00A61K39/385A61K39/102A61P31/04
Inventor 伍长华吴强关晓峰陈克平
Owner CHENGDU OLYMVAX BIOPHARM
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