猪支原体肺炎活疫苗(168株)生产工艺关键技术研究

Study on the Key Production Process of Mycoplasma hyopneumoniae, Live Vaccine (168 strain)

关键词:  猪肺炎支原体  改良培养基  发酵工艺  耐热保护剂

Key words:  Mycoplasma hyopneumoniae  live vaccine  modified medium  fermentation technology  heat resisting protective

摘要:

肺炎支原体的培养和保存条件非常苛刻，是疫苗规模化生产的工艺难题。本文针对猪支原体肺炎活疫苗（168株）生产工艺关键技术进行研究。通过培养试验筛选四种培养基配方，CCU测定证明该疫苗株在低血清改良培养基中生长良好；优化发酵培养工艺，使其在发酵罐培养60~70 h的峰值可达到1010 CCU/mL；设计筛选168株耐热保护剂和冻干工艺，37℃下保存10d的耐老化试验结果显示，下降滴度小于10 0.5 CCU/mL。本研究为提供高效、安全和稳定的猪肺炎支原体疫苗产品奠定基础。

Abstract:

It is strictly hard for culturing and storage of Mycoplasma hyopneumoniae (Mhp),which is the technical problem of vaccine integrated production. We screened four kind of culture medias of Mhp, and the color-changing units (CCU) detection results showed that Mhp strain 168 grew well in low-serum modified medium. For the optimization of the fermentation technology, the peak titers of highest growth, of strain 168, were 1010CCU /mL after cultured 60~70h in fermentor. With optimization of heat resisting protective and lyophilization technology, the titers of strain 168 were only reduced 100.5CCU /mL during the aging tolerance test which lasted for 10 days at condition of 37℃. This study built the foundation for the efficient, safety and stabile producing the Mhp live vaccines.