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Corynebacterium diphtheriae culture medium and method for preparing diphtheria toxoid by applying same

A diphtheria bacillus and culture medium technology applied in the biological field to achieve the effects of simple operation, avoiding potential hazards, and easy material collection

Inactive Publication Date: 2015-01-07
CHENGDU OLYMVAX BIOPHARM
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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These ingredients have certain potential risks

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  • Corynebacterium diphtheriae culture medium and method for preparing diphtheria toxoid by applying same
  • Corynebacterium diphtheriae culture medium and method for preparing diphtheria toxoid by applying same
  • Corynebacterium diphtheriae culture medium and method for preparing diphtheria toxoid by applying same

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] 1. Preparation of diphtheria culture medium

[0031] The culture medium is composed of basal culture medium, No. 1 solution and No. 2 solution, and the weight ratio is 100:0.3:0.3;

[0032] The basal medium is composed of raw materials of the following concentrations: HyPep5630: 10g / L, Amisoy: 10g / L, Na 2 HPO 4 12H 2 O: 1g / L, lactic acid: 2g / L, yeast powder: 0.1g / L, calcium chloride: 5g / L;

[0033] The No. 1 solution is an L-cystine solution with a concentration of 150g / L;

[0034] The No. II liquid is composed of raw materials with the following concentrations: magnesium sulfate: 100g / L, β-alanine: 0.5g / L, nicotinic acid: 0.5g / L, pimelic acid: 0.04g / L, pentahydrate Copper sulfate: 0.3g / L, zinc sulfate heptahydrate: 0.2g / L, manganese chloride tetrahydrate: 0.1g / L.

[0035] 2. The method for preparing diphtheria toxoid, it comprises the following steps

[0036] S1. Bacterial opening: open the diphtheria bacterial strain, inoculate it on the diphtheria bacillus cult...

Embodiment 2

[0042] 1. Preparation of diphtheria culture medium

[0043] The culture medium is composed of basal culture medium, No. 1 solution and No. 2 solution, and the weight ratio is 100:0.6:0.6;

[0044] The basal medium is composed of the following concentrations of raw materials: HyPep5630: 20g / L, Amisoy: 20g / L, Na 2 HPO 4 12H 2 O: 5g / L, lactic acid: 3g / L, yeast powder: 0.5g / L, calcium chloride: 15g / L;

[0045] The No. 1 solution is an L-cystine solution with a concentration of 250g / L;

[0046] The No. II liquid is composed of raw materials with the following concentrations: magnesium sulfate: 120g / L, β-alanine: 1.5g / L, nicotinic acid: 1.5g / L, pimelic acid: 0.15g / L, pentahydrate Copper sulfate: 0.8g / L, zinc sulfate heptahydrate: 0.8g / L, manganese chloride tetrahydrate: 0.2g / L.

[0047] 2. The method for preparing diphtheria toxoid, it comprises the following steps

[0048] S1. Bacterial opening: open the diphtheria bacterial strain, inoculate it on the diphtheria bacillus cul...

Embodiment 3

[0054] 1. Preparation of diphtheria culture medium

[0055] The culture medium is composed of basal culture medium, No. 1 solution and No. 2 solution, and the weight ratio is 100:0.4:0.5;

[0056] The basal medium is composed of the following concentrations of raw materials: HyPep5630: 13g / L, Amisoy: 13g / L, Na 2 HPO 4 12H 2 O: 1.5g / L, lactic acid: 2.3g / L, yeast powder: 0.25g / L, calcium chloride: 6g / L;

[0057] The No. 1 solution is an L-cystine solution with a concentration of 180g / L;

[0058] The No. II liquid is composed of raw materials with the following concentrations: magnesium sulfate: 105g / L, β-alanine: 0.8g / L, nicotinic acid: 0.8g / L, pimelic acid: 0.05g / L, pentahydrate Copper sulfate: 0.4g / L, zinc sulfate heptahydrate: 0.3g / L, manganese chloride tetrahydrate: 0.13g / L.

[0059] 2. The method for preparing diphtheria toxoid, it comprises the following steps

[0060] S1. Bacterial opening: open the diphtheria bacterial strain, inoculate it on the diphtheria bacillu...

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Abstract

The invention discloses a corynebacterium diphtheriae culture medium. The corynebacterium diphtheriae culture medium consists of a basic culture medium, an I liquid and an II liquid, wherein the basic culture medium consists of the materials of HyPep5630, Amisoy, Na2HPO4.12H2O, lactic acid, yeast powder and calcium chloride; the I liquid is L-cystine solution; the II liquid consists of the materials of magnesium sulfate, beta-alanine, niacin, pimelic acid, copper sulfate pentahydrate, zinc sulfate heptahydrate and manganese chloride tetrahydrate. The method for preparing diphtheria toxoid by applying the corynebacterium diphtheriae culture medium is culture enabling, two-generation culturing, three-generation culturing, four-generation culturing and fermentation tank culturing. The culture medium is the animal-free culture medium; good culture recovery and multiplication culture are carried out during culture passage; and the toxin production can also reach the requirement of the existing pharmacopoeia criterion. The method for preparing the diphtheria toxoid by using the culture medium has the advantages of being simple to operate, convenient to produce, easy in taking of materials and low in cost.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a diphtheria bacillus culture medium and a method for preparing diphtheria toxoid by using it. Background technique [0002] Diphtheria, an acute respiratory infection caused by the exotoxin-producing Corynebacterium diphtheriae, which can cause respiratory disturbances and even suffocation, has historically been one of the most feared childhood diseases, with devastating outbreaks. In 2006, the World Health Organization (WHO) reported that before the extensive use of diphtheria toxoid in the 1980s, there were about 1 million cases and 50,000 to 60,000 deaths in developing countries every year. Even in recent years, in endemic areas, the reported case fatality rate of diphtheria is still more than 10%. [0003] Humans are the only natural host of diphtheria bacteria. Diphtheria is only transmitted by droplets and close contact, so it is important to maintain high vaccin...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12P21/00C12R1/16
CPCC12N1/20C07K14/34
Inventor 马礼耕陈元芬
Owner CHENGDU OLYMVAX BIOPHARM
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