Simplifying CHO Fed-Batch and Perfusion Culture Media Evaluation from Expansion to Production

Today’s modern CHO based production processes employ multiple technologies to achieve optimal results. Increasingly, this includes a mix of fed-batch and perfusion-based cell culture operations. For instance, there are benefits to utilizing perfusion for seed train expansion as it allows higher cell density inoculation in production bioreactors. It also permits increases in capacity while reducing footprint.

Though providing much needed flexibility and efficiency, a mix of operations can make some aspects of the production process more complex. One area where this is particularly true is cell culture media evaluation, as different process steps and culture types require different media for best performance. Perfusion media should be optimized for perfusion processes, as fed-batch media does not support these unique requirements, such as low cell specific perfusion rates (CSPR) or high volumetric productivities In these situations, there is no “one size fits all” media choice.

Media evaluation is critical to successful cell culture from expansion to production. However, the process of media evaluation and selection can be complex, time consuming and usually requires a large range of skills and expertise. Finding the best media for the cell line, application and culture method at each process step requires thorough assessment.

Streamlined Media Evaluation Kit for Expansion through Production

To make media evaluation more efficient, MilliporeSigma has launched the 4CHO media sample kit to streamline media evaluation and provide easy to implement solutions at each stage of the production process. All the components of the kit are compatible by design so there is no need to adapt cells.

This medium kit can be used to improve current fed-batch processes or for considering perfusion as an alternative or in augmenting existing workflows. It supports evaluation from cell expansion to production in fed-batch or perfusion mode and provides quick access to evaluating the MilliporeSigma media platform. To assist in evaluating complex workflows, the basal media formulations support a diverse group of CHO cell lines, have neutral pH, require simple hydration and are paired with concentrated feeds. In addition, all components of the kit are compatible by design, so there is no need to adapt cells.

Media for all stages of the production process

The medium kit includes six samples for R&D evaluation purposes with compatible media and feeds (Figure 1).

Media kit provides evaluation options across the production process.
Figure 1: Media kit provides evaluation options across the production process.

Perfused Seed Train and Cryopreservation

The kit contains Cellvento® 4CHO-Expansion Medium designed to support cell growth at low cell specific perfusion rates (CSPR) and was developed for perfused seed train applications to spike high cell densities prior to production bioreactor innoculation. The medium can be used from vial thaw, through the entire expansion process and offers enhanced performance in subsequent fed-batch or perfusion production processes. The medium can also be used as a cryomedium when supplemented with DMSO.

Fed-batch Applications

For fed-batch processes, the kit contains EX-CELL® Advanced medium plus EX-CELL® Advanced Feed as well as Cellvento® 4CHO plus Cellvento® 4Feed. Basal media EX-CELL® Advanced and Cellvento® 4CHO can be combined with Cellvento® 4Feed or a feed mix. Cellvento® 4Feed offers the benefit of higher concentration and incorporation of modified amino acids.

Protocols for evaluating various fed-batch systems are included with the kit. One option includes the use of Cellvento® 4CHO plus Cellvento® 4Feed with a recommended culture and feed strategy. Another option includes EX-CELL® Advanced medium plus EX-CELL® Advanced Feed with a recommended culture and feed strategy. As it is recommended to evaluate several different mix levels to find the optimal feed level for each diverse cell line, a feed mixture protocol is included.

Perfusion Applications

For perfusion applications, the kit also includes EX-CELL® Advanced HD Perfusion medium to support perfusion in production scale. This medium can reach and maintain high cell densities at low CSPR, while supporting high volumetric productivities, and is designed to support the nutrient requirements of perfusion processing.

To learn more and receive a sample kit, please visit 4CHO media sample kit.

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