The pitched-blade system is traditionally the choice for stirred tank bioreactors in large-scale production of animal cells in suspension culture. The packed-bed basket technology developed by New Brunswick Scientific provides a shear-free environment for large-scale (up to 100 L) production of animal cells. At present, little information is available on the utility of this system for the production of secreted proteins, especially in perfusion mode of operation. The perfusion process provides a homeostatic environment for optimal cell growth similar to that experienced by cells in vivo. In contrast, the batch culture approach does not appropriately model this homeostatic environment due to the depletion of nutrients and accumulation of waste products in the culture system. Thus, the objective of this study was to compare the growth and productivity of alkaline phosphatase (ALKP)-secreting Chinese hamster ovary (CHO) cells cultured in these two bioreactor types: pitched-blade bioreactors operated in batch mode versus packed-bed bioreactors operated in perfusion mode. CHO cells cultured in the packed-bed bioreactor, operated in perfusion mode, produced greater amounts of ALKP compared to cells cultured in the pitched-blade system run in batch mode. These observations suggest that continuous exposure of cells to fresh culture media and the shear-free culture environment provided by the Fibra-Cel growth matrix disks offered more favorable growth conditions for CHO cells, allowing for either greater cell proliferation (higher density) or greater protein production on a per-cell basis. Overall, the results of this comparison study suggest that packed-bed bioreactors provide significant advantages for moderate-scale production of cells. The benefits of this bioreactor system may translate to large-scale cell culture for generating secreted protein products useful in medical applications…
Tag: <span>cho cells</span>
In the last few decades, laboratory and therapeutic applications of cell culture-derived biologicals have expanded from their use in diagnostic and research fields to the prevention and treatment of infectious diseases, certain forms of cancer, immunological and congenital conditions, and cell and gene therapy. While significant therapeutic benefits obtained from the use of cell culture-derived biologics (e.g., recombinant proteins, monoclonal antibodies [mAb], and vaccines) are unequivocal, the complexities associated with the manufacture of such products is acknowledged. Primary and continuous cell lines used in the manufacture can be associated with risk of contamination with endogenous retroviruses, latent viruses, or new and emerging agents. Some cell lines, such as Chinese hamster ovary (CHO) cells, have an excellent safety record with no documented safety risks…
Human granulocyte colony-stimulating factor (GCSF) is produced by biotech laboratories and production facilities for reducing neutropenia duration and sequels in patients with myelosuppressor treatments, among other applications. However, real challenges for these laboratories started in 2015 when the PEGylated-GCSF patent expired, opening alternatives for developing biomanufacturing processes and new applications. Thus, the purpose of this study was to analyze downstream process controls designed to ensure recombinant human GCSF (rh-GCSF) quality and to provide some evidence of the downstream process validation status. Study outcomes proved that the rh-GCSF expression system was stable and chromatographic profiles were reproducible among samples.
Cation exchange chromatography is typically utilized in bind-and-elute mode for monoclonal antibody purification. However, during purification process development for a novel monoclonal antibody (MAb) intended for clinical use, it was determined that bind-and-elute conditions were not sufficient for removing significant levels of antibody aggregate. Based on preliminary purification data, an alternative purification method, operation of the cation exchange process in flow-through mode, was investigated.
Raman spectroscopy offers an attractive solution for monitoring key process parameters and predictive modelling in cell culture processes using transgenic Chinese hamster ovary (CHO) cells. Frequent in-line measurements offer the potential for advanced control strategies. However, an erroneous value created by analytical signal noise is a significant issue that can affect process controls negatively. One such challenge is to differentiate the signal reflecting process changes, ranging from random to gross error, in a timely manner so the process control system can respond to these changes and maintain adequate control.
Cells cultured in 2D can differ in terms of both physiology and cellular responses compared with cells in vivo. This has led to a surge in the popularity of using 3D culture techniques as mounting evidence suggests that culturing cells in 3D is more representative of the in vivo environment, even to the extent that the gene expression profiles of cells from 3D cultures more accurately reflect clinical expression profiles than those observed in 2D cultures. 3D culture offers the potential for more accurate models of drug delivery and efficacy, as well as numerous clinical and research applications, and is becoming increasingly capable of integrating into high-throughput activities. Spheroids, or sphere cultures, have become an especially exciting area of 3D in vitro culture due to their great potential for use in studies that investigate growth and function of both malignant and normal tissues. These sphere cultures have contributed considerably to our knowledge of cellular responses thanks to the accuracy with which they reflect the in vivo system.