A novel high throughput, small scale spin tube cell culture system, which allows many hundreds of experiments to be performed in parallel, enables rapid process development such as parameter optimization, medium screening and development, cell line selection to be performed.
Data generated using this technology is predictive and scalable through shaken square bottles and shake flasks to bioreactors at 200L scale. Final processes are delivered together with all appropriate documentation needed for direct transfer to a GMP manufacturing facility for clinical lot production. Hereunder three case studies that merit the spin tube technology are described.
Selection of stable, protein expressing cell lines based on functionality. The selection of a stable cell line expressing a protein of choice is typically performed based initially on yield. For example 200 cell clones are picked, screened for protein production in a series of small scale well plates, before approx. 20 cell clones are selected for analysis in shake flask batch and/or fed-batch cultures.
The use of tube spin technology combined with high throughput purification technologies makes it possible to screen all 200 clones not only for yield but for other characteristics. These can for example be functionality based (eg: receptor binding or other in-vitro potency assays) or other important parameters such as aggregation (by HP-SEC analysis) or glycan profile (by HPLC analysis).
Medium development programs. High throughput culture system can be combined with Design of Experiment (DOE) approaches to accelerate and expand medium development programs such that many different medium components can be rapidly screened. For example a medium development program was performed on a customer mammalian cell line to develop an optimized cell culture and production medium for the manufacture of a viral vaccine product.
The program involved the screening of multiple medium components in a full factorial design of experiment approach. In total, 64 medium combinations were screened in duplicate and the data used to determine a new series of medium variants for screening in a second round. Optimal medium variants were then selected for confirmation in spinner flask and 2L bioreactors.
The advantage of such a system is that it allows duplicate or triplicate cultures to be performed, increasing the statistical relevance of the study. The volume of each culture (at 10-20ml) also allows sufficient sample volume for multiple analyses including for example metabolite analysis such that relevant decisions can be taken on medium design.
Protocol optimization. Tube spin has proven to be an effective scale down model for processes performed in larger scale bioreactors for cell lines including CHO, 293, VERO and MDCK. As such Batavia Bioservices has used the technology to perform process optimization studies in which the effect of certain parameters on product yield can be investigated.
Moreover, by combination with high throughput purification technologies, the effect of these process optimization steps on product quality (glycosylation, in-vitro efficacy, aggregation etc) can be monitored. As such, it forms a valuable tool for Quality by Design based process development and a valuable first step in setting of the design space in which the effect of the process on key quality attributes of the product can be determined.