Bioprocess Control in Microscale: Scalable Fermentations in Disposable and User-Friendly Microfluidic Systems
Bioprocess Control in Microscale: Scalable Fermentations in Disposable and User-Friendly Microfluidic Systems
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DOI：
10.1186/1475-2859-9-86
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发表时间：
2010-11-13
影响因子：
6.4
通讯作者：
Buechs, Jochen
中科院分区：
文献类型：
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作者：
Funke, Matthias;Buchenauer, Andreas;Buechs, Jochen
背景资料：生物技术生产过程的效率取决于选择性能最佳的微生物菌株和最佳的培养条件。因此，必须进行许多实验，这与加快药物开发进程的要求相冲突。因此，非常需要允许快速和可靠的生物过程开发的高通量装置。这种需要例如通过光纤在线监测系统BioLector来解决，该系统利用振荡微量滴定板（MTP）的威尔斯孔作为小规模发酵罐。为了进一步提高微生物反应器的应用，本文将BioLector技术与微流控生物过程控制相结合。为了实现一个用户友好的系统，用于常规的实验室工作，一次性的微流控MTP利用这是由一个用户友好的气动hardware.Results驱动：这种新型的微发酵系统进行了测试，在pH值控制的批次，以及在流加发酵的大肠杆菌。通过向每个培养孔中连续加入氨溶液和磷酸，可以将培养液中的pH值保持在pH设定点附近的0.03的窄死区内。此外，进行了500 g/L葡萄糖溶液的线性和指数补料的补料分批培养。最后，通过将获得的结果与在2L实验室规模发酵罐（工作体积为1 L）中完全控制的发酵的结果进行比较，评价微规模发酵的放大潜力。通过保持体积传质系数k（L）恒定在460 l/h的值来实现放大。E.结论：在微流控MTP中，成功地进行了pH控制的分批发酵以及补料分批发酵。液体剂量以及生物量的生长动力学的过程控制发酵同意以及在微型和实验室规模。总之，可以建立一种用户友好的一次性微流体系统，其允许在低于1毫升的工作体积中进行可缩放的、完全受控的和完全监测的发酵。
Background: The efficiency of biotechnological production processes depends on selecting the best performing microbial strain and the optimal cultivation conditions. Thus, many experiments have to be conducted, which conflicts with the demand to speed up drug development processes. Consequently, there is a great need for high-throughput devices that allow rapid and reliable bioprocess development. This need is addressed, for example, by the fiber-optic online-monitoring system BioLector which utilizes the wells of shaken microtiter plates (MTPs) as small-scale fermenters. To further improve the application of MTPs as microbioreactors, in this paper, the BioLector technology is combined with microfluidic bioprocess control in MTPs. To realize a user-friendly system for routine laboratory work, disposable microfluidic MTPs are utilized which are actuated by a user-friendly pneumatic hardware.Results: This novel microfermentation system was tested in pH-controlled batch as well as in fed-batch fermentations of Escherichia coli. The pH-value in the culture broth could be kept in a narrow dead band of 0.03 around the pH-setpoint, by pneumatically dosing ammonia solution and phosphoric acid to each culture well. Furthermore, fed-batch cultivations with linear and exponential feeding of 500 g/L glucose solution were conducted. Finally, the scale-up potential of the microscale fermentations was evaluated by comparing the obtained results to that of fully controlled fermentations in a 2 L laboratory-scale fermenter (working volume of 1 L). The scale-up was realized by keeping the volumetric mass transfer coefficient k(L)a constant at a value of 460 1/h. The same growth behavior of the E. coli cultures could be observed on both scales.Conclusion: In microfluidic MTPs, pH-controlled batch as well as fed-batch fermentations were successfully performed. The liquid dosing as well as the biomass growth kinetics of the process-controlled fermentations agreed well both in the microscale and laboratory scale. In conclusion, a user-friendly and disposable microfluidic system could be established which allows scaleable, fully controlled and fully monitored fermentations in working volumes below 1 milliliter.