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Method for inactivating bacterium exotoxin antigen

An exotoxin and inactivation technology, which is applied in the field of exotoxin antigen inactivation and bacterial exotoxin antigen inactivation, can solve the problems of long detoxification time and incomplete detoxification effect, so as to retain immunogenicity and improve detoxification efficiency Effect

Inactive Publication Date: 2016-06-22
LIAONING CHENGDA BIOTECH
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  • Summary
  • Abstract
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AI Technical Summary

Problems solved by technology

Formalin is the most commonly used inactivation of bacterial exotoxins, but detoxification with formalin alone has the disadvantages of long detoxification time and incomplete detoxification effect for some bacterial exotoxins

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0018] Subculture and amplification of strains: Pseudomonas aeruginosa was inoculated from the NA slant stored at 2-8°C to a fresh NA slant, and cultured at 37°C for 1 day. Pick colonies from the cultured blood plate and inoculate them into shake flasks containing fermentation medium, culture at 100-300r / min, 32-35°C for 1-2 days. The bacterial concentration of the obtained seed solution was detected by spectrophotometer method after sampling from the shake flask obtained from cultivation, and was used for fermenter cultivation after passing Gram staining microscope inspection.

[0019] Fermentation tank culture: composition of fermentation medium, beef extract 9~17g / L, bacteriological peptone 17~26g / L, sodium glutamate 6~12g / L, NaCl3~8g / L, glycerin 4~15ml / L , glucose 1~4g / L, the beef extract and bacteriological peptone were prepared in proportion to make the basal medium, which was concentrated by ultrafiltration in a 1kd membrane bag. Add sodium glutamate, sodium chloride, ...

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Abstract

The invention relates to a method for inactivating bacterium exotoxin antigen. The method is characterized by including the steps of strain passage and amplification, fermentation tank culture, exotoxin separation and purification, exotoxin inactivation and toxoid purification. Formalin, lysine and hydrogen peroxide are all used as detoxification agents, exotoxin detoxification can be completed within a shorter period of time, detoxification efficiency is improved, the immunogenicity of toxoid is kept to a larger extent, and the method has the advantages of being reliable in detoxification effect, little in influence on exotoxin structure, and capable of being widely used for detoxifying bacterium exotoxin and preparing toxoid antigen.

Description

technical field [0001] The invention relates to a method for inactivating exotoxin antigens, in particular to a method for inactivating bacterial exotoxin antigens, and belongs to the field of biotechnology. Background technique [0002] Pseudomonas aeruginosa (P.Aeruginosa) was formerly known as Pseudomonas aeruginosa. It is widely distributed in nature and is one of the most common bacteria in soil. All kinds of water, air, skin, respiratory tract and intestinal tract of normal people have this bacteria. Exotoxin A (ExoA) secreted by Pseudomonas aeruginosa is the most important pathogenic and lethal substance. After entering sensitive cells, it is activated and exerts toxic effects, hindering protein synthesis in mammals and causing tissue necrosis, causing local or Systemic disease process. Animal models have shown that liver cell necrosis, pulmonary hemorrhage, kidney necrosis and shock can occur after injection of exotoxin A to animals, and injection of exotoxin A an...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P21/00C07K1/14C07K1/34C12R1/385
CPCC12P21/00C07K1/14C07K1/34
Inventor 周荔葆王瀛杨文腰修雪亮王琦张建戴泽文
Owner LIAONING CHENGDA BIOTECH
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