肺炎链球菌4型菌株冷冻真空干燥工艺保护剂配方的筛选

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Abstract Objective To screen the formula of the protective agent for the freezing and vacuum drying process of Streptococcus pneumoniae type 4 strain through orthogonal experiments,and to analyze the activity,genetic stability and growth characteristics of the strains under the formula,and find the long-term and stable freeze-dried protective agent formulation.Methods Streptococcus pneumoniae type 4 strain was selected as the test strain.The survival rate of the strain before and after freeze-drying was used as an evaluation index.The freeze-dried protective agent formula was determined by using a three-factor orthogonal test.After freeze-drying,the activity,genetic stability and growth characteristics of the bacteria under the formula were analyzed.Results Through orthogonal experiment,the best freeze-drying protective agent formula was determined as:5% skimmed milk powder+10% trehalose+10% sodium glutamate.The concentration of the bacterial solution before lyophilization was 2.35×108 cfu/mL.After lyophilization,the strains were stored for different times(after lyophilization,storage 90,180,360 days)to test their activity,genetic stability,and growth characteristics.The activities were 81.2%,80.4%,80.1% and 79.8%.The homology analysis was 100%,and there was no difference in growth characteristics.This formulation compatible with types of 6A,18C and 19F Streptococcus pneumoniae also have good protection under the freeze-vacuum drying process,and the survival rate of the three batches of samples at day 0 was all above 78%.Conclusion After the optimization of the freeze-dried protective agent formulation,the survival rate and stability of the strains maintain a high level,and the formulation process can be used for freeze-vacuum drying and storage of different serotypes of Streptococcus pneumoniae,providing stable and reliable strains for the production of Streptococcus pneumoniae polysaccharide vaccines.

Key words： Streptococcus pneumoniae Freeze vacuum drying Protective agent Survival rate Genetic stability

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	Articles by authors
	HAO Yi-nan
	ZHANG Jing-jing
	GUO Bing-feng
	WANG Ming-hua
	WANG Ya-ping
	WANG Bin

Cite this article:

HAO Yi-nan,ZHANG Jing-jing,GUO Bing-feng, et al. Screening of protective agent formula for freezing and vacuum drying process of Streptococcus pneumoniae type 4 strain[J]. 中国当代医药, 2021, 28(14): 22-26.

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http://dangdaiyiyao.com/EN/ OR http://dangdaiyiyao.com/EN/Y2021/V28/I14/22

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