Rehfeld Johanna S, Kuhnke Louis M, Ude Christian, John Gernot T, Beutel Sascha
Institute of Technical Chemistry Leibniz University Hannover Hannover Germany.
PreSens Precision Sensing GmbH Regensburg Germany.
Eng Life Sci. 2023 Aug 23;23(9):e2300204. doi: 10.1002/elsc.202300204. eCollection 2023 Sep.
In the field of bioprocess development miniaturization, parallelization and flexibility play a key role reducing costs and time. To precisely meet these requirements, additive manufacturing (3D-printing) is an ideal technology. 3D-printing enables rapid prototyping and cost-effective fabrication of individually designed devices with complex geometries on demand. For successful bioprocess development, monitoring of process-relevant parameters, such as pH, dissolved oxygen (DO), and biomass, is crucial. Online monitoring is preferred as offline sampling is time-consuming and leads to loss of information. In this study, 3D-printed cultivation vessels with optical prisms are evaluated for the use in upstream processes of different industrially relevant microorganisms and cell lines. It was shown, that the 3D-printed optically modified well (OMW) is of benefit for a wide range of biotechnologically relevant microorganisms and even for mammalian suspension cells. Evaluation tests with , , , and Chinese hamster ovary (CHO) cells were performed, providing highly reproducible results. Growth behavior of OMW cultures was comparable to behavior of shake flask (SF) cultivations and the signal to noise ratio in online biomass measurement was shown to be reduced up to 95.8% by using the OMW. Especially the cultivation phases with low turbidity respective optical densities below 1.0 rel.AU could be monitored accurately for the first time. Furthermore, it was demonstrated that the 3D-printed optics are transferable to different well geometries and sizes, enabling efficient biomass monitoring for individual requirements with tailor-made 3D-printed cultivation vessels in small scale.
在生物工艺开发的小型化领域,并行化和灵活性对于降低成本和缩短时间起着关键作用。为了精确满足这些要求,增材制造(3D打印)是一项理想技术。3D打印能够实现快速原型制作,并按需经济高效地制造具有复杂几何形状的个性化设计设备。对于成功的生物工艺开发而言,监测与工艺相关的参数,如pH值、溶解氧(DO)和生物量,至关重要。由于离线采样耗时且会导致信息丢失,因此在线监测更为可取。在本研究中,对带有光学棱镜的3D打印培养容器在不同工业相关微生物和细胞系的上游工艺中的应用进行了评估。结果表明,3D打印的光学修饰孔(OMW)对广泛的生物技术相关微生物甚至哺乳动物悬浮细胞都有益处。对大肠杆菌、枯草芽孢杆菌、酿酒酵母和中国仓鼠卵巢(CHO)细胞进行了评估测试,结果具有高度可重复性。OMW培养物的生长行为与摇瓶(SF)培养的行为相当,并且通过使用OMW,在线生物量测量中的信噪比降低了高达95.8%。特别是首次能够准确监测低浊度或光密度低于1.0相对吸收单位(rel.AU)的培养阶段。此外,还证明了3D打印光学元件可转移到不同的孔几何形状和尺寸,从而能够使用定制的小规模3D打印培养容器根据个体需求进行高效的生物量监测。