Development of a multiplexed microbioreactor system for high-throughput bioprocessing

Abstract

A multiplexed microbioreactor system for parallel operation of multiple microbial fermentation is described. The system includes miniature motors for magnetic stirring of the microbioreactors and optics to monitor the fermentation parameters optical density (OD), dissolved oxygen (DO), and pH, in-situ and in real time. The microbioreactors are fabricated out of poly(methylmethacrylate)(PMMA) and poly(dimethylsiloxane)(PDMS), and have a working volume of 150 microl. Oxygenation of the cells occurs through a thin PDMS membrane at the top of the reactor chamber. Stirring is achieved with a magnetic spin bar in the reactor chamber. Parallel microbial fermentations with Escherichia coli are carried out in four stirred microbioreactors and demonstrate the reproducible performance of the multiplexed system. The profiles for OD, DO, and pH compare favourably to fermentations performed in bioreactor systems with multiple bench-scale reactors. Finally, the multiplexed system is used to compare two different reactor designs, demonstrating that the reproducibility of the system permits the quantification of microbioreactor performance.

Assessment of AMBR<sup>TM</sup> as a model for high-throughput cell culture process development strategy

“…Recent literature have demonstrated availability of various designs of micro-scale reactors from a simpler standard plate with integrated sensors called Microtiter plate based bioreactors (MTB) [6][7][8][9][10][11][12] to more advanced cell culture systems such as the Stirred Mini Bioreactors (MBR) with control capabilities close to bench-top bioreactors for high throughput cell culture screening and optimization studies [13][14][15][16][17]. MBRs possess the advantage of being able to closely mirror the bench-scale bio-reactors in their monitoring and control capabilities and hence are considered as a preferred alternative vessel over shake flasks or MTBs for early stage process development.…”

Nanomechanical identification of liquid reagents in a microfluidic channel

“…Significant and intriguing applications, such as drug discovery, 29 direct monitoring of products from micro-bioreactors 22 and the study of chemical composition of microbes/cells 30 are anticipated by the future integration of more sophisticated microfluidics into our system. …”

“…22 Moreover, the BMC can be a potential candidate in analyzing cellular contents of microalgae where real-time monitoring of production of different chemicals from microalgae is required. 23 Another strongly anticipated use of the BMC is the detection of circulating microvesicles (100 nm structures shed from tumor cells) which exist in low concentration in blood but have very distinct molecular signatures.…”

Smartphone-based Wireless Operated Milliliter-scale Bioreactor

“…Szita et al. [4] utilized a GPIB interface card (PCI-GPIB, National Instruments, TX, US) for the optical measurement of four 150 μL microbioreactors where multiplexing of the signals for the LEDs and the photodetectors were done using the RS232 ports. Additional I/O card i.e.…”