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Streptococcus pneumoniae freeze-drying protective agent

A freeze-drying protective agent, a technology for Streptococcus pneumoniae, which is applied in the preservation of microorganisms, microorganisms, biochemical equipment and methods, etc., can solve the problems of slow growth in the fermentation process, mutation or degradation of Streptococcus pneumoniae species, loss of immunogenicity, etc.

Active Publication Date: 2020-10-30
SHANGHAI RONGSHENG BIOLOGICAL PHARM CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, Streptococcus pneumoniae strains can mutate or degenerate due to inappropriate storage environments, resulting in slow growth during fermentation, low capsule yield, and even loss of immunogenicity

Method used

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  • Streptococcus pneumoniae freeze-drying protective agent
  • Streptococcus pneumoniae freeze-drying protective agent
  • Streptococcus pneumoniae freeze-drying protective agent

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] Streak pneumoniae type 1 bacteria on the blood plate, culture overnight at 37°C and 5% carbon dioxide environment, inoculate the grown single colony into 30ml THB medium and culture for 5 hours, then divide into 2ml sterile EP tubes , 1ml per tube. Put it into a centrifuge at 12000rpm, centrifuge for 5min, remove the supernatant, and obtain the thalline that needs to be freeze-dried. After formulating the lyophilized protective agent according to the lyophilized protective agent formula in the table below, take 1ml of the protective agent and resuspend the bacteria in the EP tube to make a bacterial suspension, and resuspend 2 EP tubes for each lyophilized protective agent . Before freeze-drying, take one EP tube resuspended with various freeze-dried powder protective agents, and spread the plate after gradient dilution to calculate the number of colonies; resuspend each freeze-dried powder with 1ml PBS, spread gradient dilution Count the number of colonies on the pla...

Embodiment 2

[0033] Preparation of 100ml lyoprotectant: Weigh 1g of sucrose, 2g of glycerin, 1g of sorbitol, 10g of skim milk powder, 0.5g of sodium glutamate, 5g of trehalose, 0.1g of vitamin C, 1g of mannitol, 1.5g of defibrated sheep blood, After dissolving with water for injection, dilute to 100ml, and then use a 0.22μm filter to filter and sterilize.

[0034] Streptococcus pneumoniae type 1 was rejuvenated on the Columbia blood plate, and the bacterial lawn was picked and put into the freeze-dried protective agent configured above to make a bacterial suspension (bacterial density of about 10 9 ~10 10 cfu), and then divided into ampoules, each ampoule 0.3ml bacterial solution.

[0035] Take out an aliquoted ampoule, dilute the bacterial solution in an appropriate gradient, spread it on a plate, put it in a carbon dioxide incubator at 37°C for overnight culture, and count the bacteria after they grow out.

[0036] Put other ampoules in a freeze dryer to freeze-dry, take a freeze-dried...

Embodiment 3

[0040] Preparation of 100ml lyoprotectant: Weigh 5g of sucrose, 2g of glycerin, 1g of sorbitol, 12g of skim milk powder, 0.5g of sodium glutamate, 5g of trehalose, 0.1g of vitamin C, 1g of mannitol, 1.5g of defibrated sheep blood, After dissolving with water for injection, dilute to 100ml, and then use a 0.22μm filter to filter and sterilize.

[0041]Streptococcus pneumoniae type 14 was rejuvenated on the Columbia blood plate, and the bacterial lawn was picked and put into the freeze-dried protective agent configured above to make a bacterial suspension (bacterial density of about 10 9 ~10 10 cfu), and then divided into ampoules, 0.3ml bacterial solution in each ampoule.

[0042] Take out an aliquoted ampoule, dilute the bacterial solution in an appropriate gradient, spread it on a plate, put it in a carbon dioxide incubator at 37°C for overnight culture, and count the bacteria after they grow out.

[0043] Put other ampoules into a freeze dryer to freeze-dry, take a freeze-...

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Abstract

The invention discloses a streptococcus pneumoniae freeze-drying protective agent which comprises skim milk powder with the concentration of 1 wt%-20 wt% and cane sugar with the concentration of 1 wt%-10 wt%. The survival rate of thalli in a streptococcus pneumoniae freeze-drying tube prepared from the freeze-drying protective agent reaches 80% or above, the character is stable, and the freeze-drying protective agent is suitable for preparing strains for streptococcus pneumoniae vaccine production.

Description

technical field [0001] The invention relates to a composition for protecting biological products, in particular to a composition for protecting biological products during the freeze-drying process to improve the properties of biological products. Background technique [0002] Streptococcus pneumoniae is a Gram-positive bacterium that causes a variety of diseases including pneumonia, meningitis, and bacteremia. According to the statistics of the World Health Organization (WHO), children under the age of 5 die from Streptococcus pneumoniae infection worldwide every year, accounting for 11% of the total number of deaths. At present, there are 93 known serotypes of Streptococcus pneumoniae, and the distribution of serotypes varies in different regions, but the infection rate of more than 10 serotypes accounts for more than 2 / 3 of Streptococcus pneumoniae diseases. Pneumococcal vaccine has become the best way to prevent and treat pneumococcus, and it is widely used in the world ...

Claims

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Application Information

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IPC IPC(8): C12N1/04C12R1/46
CPCC12N1/04
Inventor 詹晓朱绍荣张成林
Owner SHANGHAI RONGSHENG BIOLOGICAL PHARM CO LTD
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