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BryoTechnology
Scalability
BryoTechnology offers the full range from transient, small scale production to stable process development in a GMP-amenable, several hundred litres wave™-reactor.
Fig. 1: BryoSpeed™. Rapid, transient production of small quantities for feasibility and initial quality assessments. Protein is produced in moss protoplasts which are cultured in multiwell plates. Secreted product is harvested by exchange of culture medium.
 BryoSpeed™ is the rapid production system for quick delivery of initial, high quality product samples. Technically, it is based on transiently transfected protoplasts, i.e. moss cells that have been removed their plant cell wall by enzymatic digestion. After transfection, these cells are cultured in multiwell-plates, where they secrete the produced recombinant protein to the culture medium. Repeated harvest by simple media-exchange finally yields µg-amounts of high-quality, fully glycosylated product within a timeframe of six to ten weeks.
 BryoMaster™ comprises all steps of stable cell line and process development. In contrast to BryoSpeed™, the foreign DNA, embedded in a high-performance
Fig. 2: BryoMaster™.
expression cassette, is stably integrated into the genome of the production strains. These strains, selected highest producers amongst several thousands, are genetically stable without any selection pressure.
BryoMaster™ cell lines are cultivated in a photoautotrophic fermentation process. Culture vessels range from 5L-stirred tanks for process development and feasibility production to 600L-wave™ reactors. Successfully produced pharmaceutical proteins include a growth factor (VEGF), serum proteins (HSA), peptide hormones (EPO), enzymes (Phosphatase), vaccines (HA) and a wide range of oncology mAbs (IgG1 and 4).
Fig. 3: Low host cell background. Moss culture broth (moss crude) with secreted mAb-product is compared to the same mAb, produced in CHO-cells and purified (CHO mAb). All lanes are loaded with 250ng protein.
The Downstream-process is facilitated in the BryoMaster™-system by several factors. Basically, the product is secreted into a purely mineral medium with an extremely low background of host cell proteins (Fig. 3). Therefore, the initial purity of the produced protein is around 80% and the broth is free of any animal-derived substances. After moss removal, the medium is ready for a generic downstream process as applied in conventional systems.
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