High-Throughput Micro-Bioreactors (MBR)

Designing and building a high-throughput microbioreactor (MBR) system for mammalian cells is an exceedingly complex task. To make it feasible within the budget- and time-constraints conferred by a publicly funded project, the new MBR system will be built up based on key features of the BioLector Pro System developed and marketed by m2p-labs for microbial cultivations.

The product design requirements for the new system have been defined in cooperation with the pharmaceutical companies in the consortium representing a future user base of the new system. pH regulation in the wells will be achieved through individual gassing of each bioreactor with CO2. Introduction of additional gases but CO2 will also be possible. Feeding of the cultivated cells will be accomplished through a dedicated microfluidic system integrated into the MBR or through a supporting liquid handler. Online monitoring of vital conditions of the cells will be accomplished through a combination of classical optode-technology with an integrated spectroscopic system. The potential of 2D Fluorescence Spectroscopy and Near-Infrared Spectroscopy as online monitoring tools for mammalian cell culture have already been published before. Still, most details of these promising approaches have remained uncharted territory so far. Accordingly, it will be a focus of workpackage 5 to investigate corresponding algorithms to use spectroscopy to quantify biomass and concentrations of small molecules vital for cultivations. In WP 2 the most promising technology will be integrated into the MBR.

The volumes of the microbioreactor and the throughput of the microbioreactor system are normally inversely correlated. Thus, it will be a critical design goal for the MBR system to miniaturise the culture volume and allow for powerful online analysis minimising sampling need. Miniaturised reactors present specific challenges concerning individual supply with gas and nutrition while keeping stress conferred by agitation or stirring below a critical level. Assuring a correct function of small bioreactors will require an elaborate design and detailed characterisation of the miniaturised culture vessels to grant throughput and scalability.

Harvesting soluble proteins secreted by cultured cells through batch-wise collecting of the cell supernatant can impair cell growth and create poorly reproducible culture conditions. Accordingly, one focus of the activities in work package 2 will focus on generation consumables to allow for perfusion of liquids in a continuous stream into the micro reactor vessel and out of the vessel. All of these features will be integrated into a single chip for maximum throughput and ease of use.

Finally, the developed system will be validated under real screening and culture optimisation conditions by the technology developing institutions together with the partners from the pharmaceutical industry.

Workpackage Objectives

to develop on-line and at-line detections of physical factors and components necessary for a thorough knowledge of the culture
to create a new tool for High-Throughput (HTP) process development by high performing parallel micro-bioreactors (MBR) for fed-batch or for perfusion operations
to integrate the on-line/at-line sensing tools together and with the MID, as well as feed-back controls of the MBR or of the stirred tank bioreactors for process analytical technology (PAT)

Workpackage Achievements

New advancements in high-throughput process monitoring

Technical specifications of the microbioreactor system identified

Design of the new microbioreactor system

Gassing for the microbioreactors realized

Microbioreactor suitable for mammalian cell cultivations

Fully working preliminary model of MBR

Leader and Co-Leader

Anna Kress, m2p-labs GmbH

Roland Wagner, Rentschler Biopharma

Partners

Kungliga Tekniska Hoegeskolan, Sweden