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Culture medium for preparing tetanus toxin and application thereof

A technology of tetanus toxin and culture medium, applied in the biological field, can solve the problems of long toxin production cycle and low toxin production yield, etc., and achieve the effects of being conducive to large-scale production, simple material collection, and good bacterial growth.

Inactive Publication Date: 2017-01-04
CANSINO BIOLOGICS INC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] The object of the present invention is to provide a kind of culture medium for preparing tetanus toxin, which solves the defects that the toxin production cycle is long when the tetanus toxin is prepared, the toxin production is low, and the requirements of current standards cannot be met.

Method used

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  • Culture medium for preparing tetanus toxin and application thereof
  • Culture medium for preparing tetanus toxin and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0043] Preparation of Clostridium tetani culture medium

[0044] The culture medium is prepared and mixed from the base medium, the first growth factor and the second growth factor respectively; and the weight ratio is: 100:0.1:0.1; the base medium is composed of the following raw materials: yeast extract: 5g / L, peptone: 0g / L, crystalline sodium acetate: 2g / L, potassium dihydrogen phosphate: 0.5g / L, disodium hydrogen phosphate dodecahydrate: 0.5g / L, glycerin: 10ml / L, glucose: 0.5 g / L, biotin: 5 μg / L, uracil: 5 μg / L; the first growth factor is composed of the following raw materials: Nicotinic acid: 0.05g / L, cystine: 15g / L, hydrochloric acid: 20ml / L , zinc sulfate: 1g / L, magnesium sulfate heptahydrate: 15g / L; the second growth factor is made up of the following raw materials: thiamine: 0.05g / L, riboflavin: 0.05g / L, pyridoxine hydrochloride : 0.05g / L, calcium pantothenate: 0.2g / L, absolute ethanol: 20ml / L.

[0045] The preparation of tetanus toxin includes the following steps:...

Embodiment 2

[0050] Preparation of Clostridium tetani culture medium

[0051] The culture medium is prepared and mixed with the basic medium, the first growth factor and the second growth factor respectively; and the weight ratio is: 100:1:1; the basic medium is composed of the following raw materials: yeast extract: 25g / L, peptone: 10g / L, crystalline sodium acetate: 8g / L, potassium dihydrogen phosphate: 1.5g / L, disodium hydrogen phosphate dodecahydrate: 1.5g / L, glycerin: 150ml / L, glucose: 20g / L, biotin: 20 μg / L, uracil: 20 μg / L; the first growth factor is composed of the following raw materials: nicotinic acid: 0.15g / L, cystine: 35g / L, hydrochloric acid: 80ml / L, Zinc sulfate: 5g / L, magnesium sulfate heptahydrate: 50g / L; the second growth factor consists of the following raw materials: thiamine: 0.15g / L, riboflavin: 0.15g / L, pyridoxine hydrochloride: 0.15g / L, calcium pantothenate: 0.6g / L, absolute ethanol: 140ml / L.

[0052] The preparation of tetanus toxin includes the following steps: ...

Embodiment 3

[0057] Preparation of Clostridium tetani culture medium

[0058] The culture medium is prepared and mixed from the base medium, the first growth factor and the second growth factor respectively; and the weight ratio is: 100:0.2:0.2, and the base medium is composed of the following raw materials: yeast extract: 7.5 / L, peptone: 0g / L, crystalline sodium acetate: 3g / L, potassium dihydrogen phosphate: 0.8g / L, disodium hydrogen phosphate dodecahydrate: 0.8g / L, glycerin: 30ml / L, glucose: 1g / L, biotin: 8μg / L, uracil: 7.5μg / L; the first growth factor is composed of the following raw materials: Nicotinic acid: 0.065g / L, cystine: 20g / L, hydrochloric acid: 35ml / L , zinc sulfate: 2g / L, magnesium sulfate heptahydrate: 20g / L; the second growth factor consists of the following raw materials: thiamine: 0.08g / L, riboflavin: 0.08g / L, pyridoxine hydrochloride : 0.08g / L, calcium pantothenate: 0.3g / L, absolute ethanol: 40ml / L.

[0059] The preparation of tetanus toxin includes the following step...

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Abstract

The invention discloses a culture medium for preparing tetanus toxin and a method for preparing the tetanus toxin from the culture medium. The culture medium contains an organic nitrogen source of a yeast source, the mass percent of the organic nitrogen source of an animal source and / or a plant source in the organic nitrogen source of the culture medium is smaller than 1-50%. According to the culture medium, the risk of introducing zoonotic diseases such as mad cow disease is avoided, strain anabiosis and seed culture can be met, and produced toxin is also far higher than the requirements of the existing standard. The operation of preparing the tetanus toxin by utilizing the culture medium is simple, the cost is low, and the method is favorable for industrial production.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a culture medium for preparing tetanus toxin and its use for seed culture and fermentation to prepare tetanus toxin. Background technique [0002] Tetanus is a disease with high mortality caused by Clostridium tetani. The global incidence is not easy to determine, but it is estimated that 400,000 to 1 million people are infected with tetanus every year. There is no medical evidence to prove that there is an effective treatment option, but for most people, the use of vaccines can provide effective protection. [0003] Tetanus is an infectious disease, not a contagious disease. Tetanus infects wounds, and produces tetanus neurotoxin (referred to as tetanus toxin) under hypoxic conditions, which is brought into the blood system through the blood and diffuses to the nervous system throughout the body. The body absorbs it into the nervous system and acts on the central nervou...

Claims

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Application Information

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IPC IPC(8): C12P21/02
Inventor 代帆朱涛史建明常云松宇学锋邵忠琦毛慧华
Owner CANSINO BIOLOGICS INC
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