A versatile, non genetically modified organism (GMO)-based strategy for controlling low-producer mutants in Bordetella pertussis cultures using antigenic modulation
A versatile, non genetically modified organism (GMO)-based strategy for controlling low-producer mutants in Bordetella pertussis cultures using antigenic modulation
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DOI：
10.1002/biot.201400539
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发表时间：
2015-08-01
影响因子：
4.7
通讯作者：
Dehottay, Philippe
中科院分区：
文献类型：
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作者：
Goffin, Philippe;Slock, Thomas;Dehottay, Philippe
非生产者突变体的不受控制的存在对生物过程产生负面影响。在百日咳博德特氏菌培养物中，无毒力突变体自发出现并积累。我们使用高通量生长试验和强毒和无毒（bvg（-））分离株之间的竞争实验来表征积累的动态。在高毒力因子生产条件下，bvg（-）细胞的适应性优势被确定为bvg（-）积累的主要驱动力。相反，在减少其表达（抗原调节）的条件下，可以避免bvg（-）接管。一种控制策略，它包括在应用调制条件时，毒力因子的生产是不需要的。它具有广泛的应用范围，从常规实验室操作到疫苗生产，其中通过在调节条件下进行早期预培养步骤，百日咳毒素产量增加了1.4倍。因为它只需要对培养基进行细微的修改，不涉及基因修饰，所以这种策略适用于任何B。百日咳分离株，并应促进疫苗生产工艺变更的监管接受。基于同一概念的战略，可用于其他工业相关微生物。这项研究说明了如何将对生理学原理的合理科学理解转化为生物工艺行业的实际应用，并与设计质量原则保持一致。
The uncontrolled presence of non-producer mutants negatively affects bioprocesses. In Bordetella pertussis cultures, avirulent mutants emerge spontaneously and accumulate. We characterized the dynamics of accumulation using high-throughput growth assays and competition experiments between virulent and avirulent (bvg(-)) isolates. A fitness advantage of bvg(-) cells was identified as the main driver for bvg(-) accumulation under conditions of high virulence factor production. Conversely, under conditions that reduce their expression (antigenic modulation), bvg(-) takeover could be avoided. A control strategy was derived, which consists in applying modulating conditions whenever virulence factor production is not required. It has a wide range of applications, from routine laboratory operations to vaccine manufacturing, where pertussis toxin yields were increased 1.4-fold by performing early pre-culture steps in modulating conditions. Because it only requires subtle modifications of the culture medium and does not involve genetic modifications, this strategy is applicable to any B. pertussis isolate, and should facilitate regulatory acceptance of process changes for vaccine production. Strategies based on the same concept, could be derived for other industrially relevant micro-organisms. This study illustrates how a sound scientific understanding of physiological principles can be turned into a practical application for the bioprocess industry, in alignment with Quality by Design principles.