…your decision may depend on cell culture technology
The transient expression of monoclonal antibodies is an essential step for early characterization studies but to produce 500 – 1000 mg of mAb requires a huge quantity of plasmid and transfection agent. The inherent heterogeneity of the cells means there’s no guarantee that post-translational modifications will be identical or maintained when repeating the process. This disadvantage might significantly impact translational research projects, for example.
Advances in technology are rendering the generation of stably expressing pools of cells and subsequent clone selection much easier than in the past. Combining stable expression with the use of a compact high cell density culture system enables any modestly equipped laboratory to conduct repeated gram-scale productions over many years with no batch-to-batch variation. A well-designed hollow fibre bioreactor is the best solution for such laboratories. Read more about hollow fibre technology here : https://www.kdbio.com/fibercell-hollow-fibre-technology/
For example, a FiberCell bioreactor will comfortably fit inside a standard 180L incubator and can be purchased on a small equipment budget. Around 50 mg per week of concentrated recombinant protein can be generated from a 70ml cartridge over a 3 to 6 month period. A single bioreactor pump unit will accommodate two such cartridges which are conveniently purchased as sterile gamma-irradiated, double-bagged, single-use consumables with all tubing and connections included.
Transient transfections may require specialised and costly media. When growing cells in a Fibercell bioreactor the use of a simplified, economical chemically defined media is sufficient because natural cell-secreted growth factors are present at high concentration. To sustain cell growth without serum or complex media the recommended media would be CDM-HD. This is a powder that is made up to 1 litre with distilled water and added at 10% to basal DMEM. Only around 350 grams are needed to produce 100 Litres of final bioreactor media.
As well as media simplification, the stress-free conditions in a hollow fibre bioreactor are favourable to achieving complete and uniform post-translational modifications. This topic is explained in more detail by John Cadwell, CEO of FiberCell Systems in this article: A hollow fibre bioreactor enables any lab to efficiently express recombinant proteins in mammalian cells
Example:
Increased phosphorylation of HexM improves lysosomal uptake and potential for managing GM2 gangliosidoses: Benzie, G. et al. BBA Advances Vol.2, 2022 [open access]
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