基于质量源于设计理念在伤寒沙门菌中试规模培养工艺放大中的应用

The application of quality by design concepts in technological scale-up of pilot culture of Salmonella typhi

目的基于质量源于设计(quality by design, QbD)理念,确定伤寒沙门菌(Salmonella typhi)中试规模(200 L)的培养工艺。方法运用QbD通过分析伤寒沙门菌培养工艺核心指标、培养过程,以确定对核心指标可能产生影响的因素。采用单因素试验对pH控制值、溶氧控制值和补料方式等进行考察,通过中试规模(20 L)培养工艺桥接和中试规模(200 L)培养工艺的放大,分析中试规模培养条件及发酵液中伤寒Vi荚膜多糖含量等。结果连续培养了6批次200 L规模发酵液,培养8 h时发酵液中伤寒Vi荚膜多糖含量均≥43.33μg/mL(期望值);培养条件:(1) pH控制值为7.2;(2)溶氧控制值为35%;(3)补料方式为培养初始补加葡萄糖溶液使其质量浓度为3 g/L,培养过程中补加葡萄糖溶液使其质量浓度保持在1~3 g/L。结论通过分析200 L规模培养过程监测数据,成功在中试规模(200 L)完成伤寒沙门菌培养工艺放大。

Objective To determine the pilot scale(200 L) culture process of Salmonella typhi. Methods Based on the concept of quality by design(QbD), the core indicators in the process of Salmonella typhi culture were analyzed.The factors that affect these core indicators obtained by analyzing the cultivation process. A single factor test method was used to investigate the factors such as pH control value, dissolved oxygen control value and feeding method. Based on the pilot scale(20 L) culture process bridging experiments and 200 L culture process, pilot scale culture conditions and typhoid Vi capsular polysaccharide content in batch fermentation broth were analyzed. Results 6 batches of 200 L scale culture were conducted, the content of typhoid Vi capsular polysaccharides in the batch fermentation broth was more than 43.33 ug/mL(the expected value) at 8 h;culture conditions were as follows :(1) pH control value was 7.2;(2) dissolved oxygen control value was 35%;(3) feeding method: the initial glucose concentration was 3 g/L, and maintained the concentration at(1-3)g/L during the cultivation. Conclusion By analyzing the monitoring data of the 200 L scale culture process, the fermentation culture of Salmonella typhi was successfully scaled up in a pilot scale(200 L).