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Preparation method of group A meningococcus capsular polysaccharide conjugate

A technology of meningococcal and capsular polysaccharides, applied in the field of vaccine conjugates, can solve the problems of long production cycle, cumbersome preparation process, and long time consumption, and achieve increased stability, simplified preparation process, polysaccharide yield and protein yield high effect

Pending Publication Date: 2021-12-03
ZHEJIANG VACIN BIO PHARMA LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

As a result, the entire preparation process of the group A meningococcal capsular polysaccharide conjugate is cumbersome and time-consuming, which in turn makes the production cycle of the group A meningococcal conjugate vaccine longer

Method used

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Examples

Experimental program
Comparison scheme
Effect test

preparation example Construction

[0039] Examples of Preparation of Raw Materials and / or Intermediates

[0040] Extraction of Capsular Polysaccharide of Group A Meningococcus

[0041] The strains of group A meningococcus were purchased from a strain bank, and after subculture for 6 generations, the culture solution containing group A meningococcus was harvested. Take 1L of the above-mentioned culture solution and sterilize it with formaldehyde to ensure the safety of sterilization without damaging the polysaccharides of the bacteria. Centrifuge the sterilized culture solution to remove bacteria, collect the supernatant, concentrate the harvested supernatant to 1 / 8 of the original volume with an ultrafiltration membrane with a molecular weight cut-off of 50-100KD, and add hexadecyltrimethyl bromide Mix with ammonium chloride, stir at room temperature for 60 minutes, let stand at 4°C for 8 hours, collect the precipitate by high-speed centrifugation at 15,000 r / min, add calcium chloride solution to the precipita...

Embodiment 1

[0046] A method for preparing a group A meningococcal capsular polysaccharide conjugate, comprising the following steps:

[0047] ①. Preparation of polysaccharide derivatives

[0048] Weigh 1.0 g of group A meningococcal capsular polysaccharide, put it in a beaker, add 200 mL of normal saline (25°C), stir with a magnetic stirrer at room temperature for 30 min at a stirring speed of 400 r / min, and dissolve it into polysaccharide solution a;

[0049] Use a sodium hydroxide solution with a molar concentration of 0.1mol / L to adjust the pH of the polysaccharide solution a to 10.5, and the pH is allowed to fluctuate in the range of ±0.2 (fluctuations within this range have less impact on product performance), adding a mass concentration of Activate 1.2L of 1g / L hydrogen bromide solution to obtain polysaccharide solution b;

[0050] Add 3.0L of adipic hydrazide solution with a molar concentration of 1g / L to the polysaccharide solution b to derivatize for 15min. During this period, t...

Embodiment 2-4

[0065] Embodiment 2-4 On the basis of the method in Embodiment 1, the process parameters in step ② are adjusted, and the specific adjustment is shown in Table 1 below.

[0066] Table 1 The process parameter adjustment table of embodiment 1-4

[0067] Example 1 Example 2 Example 3 Example 4 Reaction concentration of polysaccharide derivatives (mg / mL) 1.2 0.5 1.6 5.0 Weight ratio of polysaccharide derivatives to diphtheria CRM197 protein 1:1 1:0.8 1:1.5 1:2.5

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Abstract

The invention relates to the technical field of vaccine conjugates, in particular to a preparation method of a group A meningococcus capsular polysaccharide conjugate, which comprises the steps of polysaccharide derivative preparation, mixing proportioning, conjugation reaction, aging treatment, ultrafiltration and sterilization. According to the method, the processes of pH adjustment in the activation process and conjugate purificationare omitted through optimization of all technological parameters, but the quality of the harvested polysaccharide conjugate still meets the national standard, so that the preparation process of the polysaccharide conjugate is simplified, and the preparation period of the polysaccharide conjugate is shortened.

Description

technical field [0001] This application relates to the technical field of vaccine conjugates, more specifically, it relates to a preparation method of a group A meningococcal capsular polysaccharide conjugate. Background technique [0002] Epidemic meningitis is an epidemic of meningeal inflammation caused by Neisseria meningitidis (n.meninyitidis). The types of strains that cause human lesions are mainly group A, group B, group C, group Y and group W135. More than 90% of meningitis cases in my country are caused by group A Neisseria meningitidis. [0003] Vaccine is an effective way to prevent and control meningococcal infection. Group A meningococcal conjugate vaccine is to extract the bacterial outer capsule polysaccharide of group A strain meningococcus, and couple the capsular polysaccharide with specific protein or polypeptide to produce into a polysaccharide conjugate, and then formulated into a vaccine to prevent the invasion and infection of this strain of meningoco...

Claims

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Application Information

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IPC IPC(8): A61K39/385A61P25/00A61P31/04C08B37/00
CPCA61K39/385C08B37/00A61P31/04A61P25/00A61K2039/6037
Inventor 马伟朱楠
Owner ZHEJIANG VACIN BIO PHARMA LTD
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