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Process for activating Haemophilus influenzae type b (Hib) polysaccharide conjugate vaccine

A Haemophilus influenzae and conjugate vaccine technology is applied in the field of Haemophilus influenzae type b polysaccharide conjugate vaccine activation technology, which can solve problems such as use, and achieve the effects of avoiding harm and improving safety.

Active Publication Date: 2012-08-08
CHENGDU OLYMVAX BIOPHARM
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The disadvantage of this process is the use of cyanogen bromide in the bonding process

Method used

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  • Process for activating Haemophilus influenzae type b (Hib) polysaccharide conjugate vaccine
  • Process for activating Haemophilus influenzae type b (Hib) polysaccharide conjugate vaccine
  • Process for activating Haemophilus influenzae type b (Hib) polysaccharide conjugate vaccine

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Experimental program
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Embodiment 1

[0025] Embodiment 1: This embodiment is the best embodiment of the present invention.

[0026] Haemophilus influenzae type b polysaccharide conjugated vaccine activation process, it comprises the following steps:

[0027] A, prepare Hib polysaccharide, it comprises the following substeps:

[0028] A1. Centrifuge the sterilized culture solution at 10,000 rpm for 30 minutes to precipitate the bacteria, and collect the supernatant;

[0029] A2. Add CTAB to the supernatant, the weight-to-volume ratio (w / v) of CTAB to the supernatant is 1:1000, stir at room temperature for 1 h, and collect the composite of CTAB and Hib capsular polysaccharide after centrifuging the obtained mixture at 10,000 rpm for 30 min precipitation;

[0030] A3, the complex precipitate was dissolved with 1M sodium chloride solution, and stirred at room temperature for 2 hours to dissociate the Hib capsular polysaccharide from CTAB; add 4°C pre-cooled absolute ethanol until the final concentration of ethanol ...

Embodiment 2

[0043] Haemophilus influenzae type b polysaccharide conjugated vaccine activation process, it comprises the following steps:

[0044] A, prepare Hib polysaccharide, it comprises the following substeps:

[0045] A1. Centrifuge the sterilized culture solution at 8000 rpm for 40 minutes to precipitate the bacteria, and collect the supernatant;

[0046] A2, CTAB is added in the supernatant, the weight-to-volume ratio (w / v) of CTAB and supernatant is 1: 800, stir at room temperature for 0.5h, and the obtained mixture is centrifuged at 8000rpm for 40min to collect the mixture of CTAB and Hib capsular polysaccharide complex precipitation;

[0047] A3. Dissolve the complex precipitate with 0.5M sodium chloride solution, stir at room temperature for 1 hour to dissociate Hib capsular polysaccharide from CTAB; add 3°C pre-cooled absolute ethanol until the final concentration of ethanol is 20% (v / v) , stirred overnight at 3°C;

[0048] A4. Centrifuge at 800rpm for 40min, collect the su...

Embodiment 3

[0060] Haemophilus influenzae type b polysaccharide conjugated vaccine activation process, it comprises the following steps:

[0061] A, prepare Hib polysaccharide, it comprises the following substeps:

[0062] A1. Centrifuge the sterilized culture solution at 12000 rpm for 20 minutes to precipitate the bacteria, and collect the supernatant;

[0063] A2. Add CTAB to the supernatant, the weight-to-volume ratio (w / v) of CTAB to the supernatant is 1:1200, stir at room temperature for 1.5h, and collect the mixture of CTAB and Hib capsular polysaccharide after centrifuging at 12000rpm for 20min. complex precipitation;

[0064] A3. Dissolve the complex precipitate with 1.5M sodium chloride solution, stir at room temperature for 3 hours to dissociate Hib capsular polysaccharide from CTAB; add 5°C pre-cooled absolute ethanol until the final concentration of ethanol is 30% (v / v) , stirred overnight at 5°C;

[0065] A4. Centrifuge at 12000rpm for 20min, collect the supernatant, conti...

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Abstract

The invention discloses a process for activating a Haemophilus influenzae type b (Hib) polysaccharide conjugate vaccine, which comprises the following steps of: A, preparing Hib polysaccharide; B, dissolving 1-Cyano-4-dimethylaminopyridinium tetrafluoroborate (CDAP) by using acetonitrile into a solution; C, preparing the Hib polysaccharide into a solution; D, adding the CDAP solution to the Hib polysaccharide solution, and stirring for 2-5 minutes at the room temperature; E, dissolving adipic dihydrazide (ADH) by using NaHCO3 into a solution, adding the ADH solution to a mixed solution, and stirring for 0.5-2 hours at the room temperature; and F, collecting eluent of the void volume from a loading solution after the reaction is ended at a SephadexG-25 gel chromatography column balanced in advance by water for injection, and performing freeze drying to obtain a Hib polysaccharide-ADH derivative. The process for activating the Hib polysaccharide conjugate vaccine has the beneficial effects that the quality index of the prepared Hib polysaccharide-ADH can reach the industrial standard, and moreover, the safe and nontoxic CDAP is adopted to serve as an activating agent instead of cyanogen bromide which is greatly harmful to human and environment, and therefore, the safety is enhanced, and the harm to the human and the environment are avoided.

Description

technical field [0001] The invention relates to the technical field of polysaccharide activation methods, in particular to the activation process of Haemophilus influenzae type b polysaccharide conjugated vaccine. Background technique [0002] Haemophilus influenzae is still the main pathogenic bacteria causing human invasive diseases so far, most of which are caused by Haemophilus influenzae type b (Hib), which is the main cause of meningitis and bacterial pneumonia in infants under 2 years old . In 2006, the World Health Organization (WHO) reported that at least 3 million cases of severe diseases and 400,000 to 700,000 deaths were caused worldwide each year, which has become a major global public health problem. [0003] Hib capsular polysaccharide is one of the main virulence factors of Hib. It is a polymer composed of repeating units with phosphoribosyl ribitol (PRP) as the main component. It has good immunogenicity and can induce Produces potent protective bactericida...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K39/385A61K39/102A61P31/04C08B37/00
Inventor 吴强伍长华魏新陈爱民
Owner CHENGDU OLYMVAX BIOPHARM
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