A-group C-group Neisseria meningitidis polysaccharide conjugate vaccine activating process

Abstract

The invention discloses an A-group C-group Neisseria meningitidis polysaccharide conjugate vaccine activating process. The problem that highly-toxic chemical reagents, such as cyanogen bromide, are used in the vaccine manufacturing process in the prior art and accordingly are harmful to protection of human health and environment is solved. The A-group C-group Neisseria meningitidis polysaccharide conjugate vaccine activating process comprises the following steps of 1 cultivating Neisseria meningitidis, 2 utilizing the cultivated Neisseria meningitidis to extract crude polysaccharide, 3 purifying the crude polysaccharide to prepare refined Neisseria meningitidis polysaccharide and 4 activating and deriving the refined polysaccharide. The A-group C-group Neisseria meningitidis polysaccharide conjugate vaccine activating process has the advantages that the extraction rate of the polysaccharide from a capsule is higher, the precipitation rate of the polysaccharide is higher, the recovery rate and purity of the refined polysaccharide are high, the derivation effect is better and the like.

Description

technical field

[0001] The invention relates to a method for cultivating strains, in particular to an activation process for group A and group C meningococcal polysaccharide-conjugated vaccines. Background technique

[0002] Meningococcal meningitis (referred to as meningitis, the same below) is an acute respiratory infectious disease caused by Neisseria meningitidis. For more than one hundred years, it has been popular or sporadic all over the world. It can cause sepsis and meningitis after infection with pathogenic bacteria. The susceptible population is mainly children, and the case fatality rate of fulminant type is the highest, which can reach 40% to 60%. The incidence rate in all continents of the world today is 1 / 100,000 to 10 / 100,000, and the total case fatality rate is 5% to 15%. Up to 20% of meningitis patients will have neurological sequelae, including mental impairment and deafness. Serological classification according to the type of capsular polysaccharide can...

Examples

Embodiment 1

[0045] The activation process of group A group C meningococcal polysaccharide conjugate vaccine comprises the following steps:

[0046] (1) Cultivate meningococci;

[0047] (11) Preparation of purified working seed batch strains: Inoculate group A Neisseria meningitidis strains on the purified medium, incubate at 38°C for 20 hours, select robust and free strains of bacteria and inoculate again Carry out secondary culture on the purified medium, and then cultivate at 38°C for 12 hours, select robust and non-contaminated strains and add them to aseptic skim milk, mix and freeze-dry to prepare purified working seed batch strains ;

[0048] (12) Dissolve the working seed batch strains into sterile water, inoculate them on 10% sheep blood agar medium, and culture them at 35-37°C for 16-20 hours, then select the robust and impurity-free strains. Bacteria-contaminated strains; inoculate the opened purified seeds into a semi-comprehensive liquid medium for activation culture; the co...

Embodiment 2

[0072] The only difference between this example and Example 1 is that the weight ratio of polysaccharide to CDAP in step (42) in this example is 1:0.6; the weight ratio of polysaccharide to ADH in step (43) is 1:4.

Embodiment 3

[0074] The difference between this embodiment and embodiment 1 is only that the proportion and concentration of each chemical substance in this embodiment are added to the reaction, and the specific settings are as follows:

[0075] The concentration of the polysaccharide solution in step (41) is 10 mg / ml, and the pH is adjusted to 9.0 with sodium hydroxide;

[0076] The concentration of CDAP solution in step (42) is 80mg / ml, polysaccharide:CDAP=1:0.7;

[0077] The concentration of ADH solution in step (43) is 80mg / ml ADH solution, polysaccharide: ADH=1:5.

technical field

[0001] The invention relates to a method for cultivating strains, in particular to an activation process for group A and group C meningococcal polysaccharide-conjugated vaccines. Background technique

[0002] Meningococcal meningitis (referred to as meningitis, the same below) is an acute respiratory infectious disease caused by Neisseria meningitidis. For more than one hundred years, it has been popular or sporadic all over the world. It can cause sepsis and meningitis after infection with pathogenic bacteria. The susceptible population is mainly children, and the case fatality rate of fulminant type is the highest, which can reach 40% to 60%. The incidence rate in all continents of the world today is 1 / 100,000 to 10 / 100,000, and the total case fatality rate is 5% to 15%. Up to 20% of meningitis patients will have neurological sequelae, including mental impairment and deafness. Serological classification according to the type of capsular polysaccharide can...

Examples

Embodiment 1

[0045] The activation process of group A group C meningococcal polysaccharide conjugate vaccine comprises the following steps:

[0046] (1) Cultivate meningococci;

[0047] (11) Preparation of purified working seed batch strains: Inoculate group A Neisseria meningitidis strains on the purified medium, incubate at 38°C for 20 hours, select robust and free strains of bacteria and inoculate again Carry out secondary culture on the purified medium, and then cultivate at 38°C for 12 hours, select robust and non-contaminated strains and add them to aseptic skim milk, mix and freeze-dry to prepare purified working seed batch strains ;

[0048] (12) Dissolve the working seed batch strains into sterile water, inoculate them on 10% sheep blood agar medium, and culture them at 35-37°C for 16-20 hours, then select the robust and impurity-free strains. Bacteria-contaminated strains; inoculate the opened purified seeds into a semi-comprehensive liquid medium for activation culture; the co...

Embodiment 2

[0072] The only difference between this example and Example 1 is that the weight ratio of polysaccharide to CDAP in step (42) in this example is 1:0.6; the weight ratio of polysaccharide to ADH in step (43) is 1:4.

Embodiment 3

[0074] The difference between this embodiment and embodiment 1 is only that the proportion and concentration of each chemical substance in this embodiment are added to the reaction, and the specific settings are as follows:

[0075] The concentration of the polysaccharide solution in step (41) is 10 mg / ml, and the pH is adjusted to 9.0 with sodium hydroxide;

[0076] The concentration of CDAP solution in step (42) is 80mg / ml, polysaccharide:CDAP=1:0.7;

[0077] The concentration of ADH solution in step (43) is 80mg / ml ADH solution, polysaccharide: ADH=1:5.

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