关键词: 发酵, 细胞培养, 平行培养, 高通量, 合成生物学, 实验设计（DoE）

Abstract:

At the beginning of this century, high-throughput in-parallel fermentation (cell culture) technology and its related apparatus based on microbioreactor and miniature bioreactor were developed and widely applied. This was to solve the challenges encountered by biopharmaceutical process researches in terms of cultivation throughput, R&D efficiency, and the continuous rising on the cost for microbial fermentation and mammalian cell cultures of multiple clones, and more importantly, the urgent needs for conducting design of experiment (DoE) driven by the principle of quality by design (QbD). In the recent years, the rapid advance of microbial metabolic engineering and synthetic biology has put forward a rather strong demands for the high-throughput screening on high-performance strain libraries and the early evaluation of strain phenotypic potential performance under industrial cultivation conditions. This has further expanded the application range of in-parallel fermentation technology with various culture volumes in the field of modern industrial biotechnology. Up to now, integrated systems containing in-parallel fermentation setups with multiple microbioreactors, online probes, operating software, and data processing units, which can simulate industrial cultivation conditions and implement accurate control of process parameters, have become a powerful tool for accelerating bioprocesses engineering R&D. They play a central role in transforming basic research achievements, such as drug discovery, metabolic engineering, and synthetic biology, into industrial technologies. Especially in the field of synthetic biology, based on the principle of "industrial similarity", in-parallel fermentation technology addresses key limitations of the conventional microplate and shake flask based high throughput screening, which cannot characterize the significant impact of culture conditions on the phenotypic performance of the selected clones at a large scale. This enables a process-oriented, high throughput, and efficient screening and evaluation if microbial strain libraries. This review provides an overview on the recent development in high throughput in-parallel fermentation & cell culture technology and its application scenarios of synthetic biology researches. In particular, it emphasizes the value of in-parallel fermentation technology in high-throughput strain screening complying with the three-stage strategy, and how in-parallel fermentation technology makes the implementation of the industrial similarity principle of strain screening possible, and how the technology combines DoE experimental tactic to significantly improve the efficiency of bioprocess development. It is also discussed the general procedure to build up a multivariate batch model of bioprocess based on in-parallel fermentation technology. In the end the approach of using in-parallel culture to establish a process scale down model is also explored.

Key words: fermentation, cell culture, in-parallel cultivation, high throughput, synthetic biology, design of experiment (DoE)