Bioreactor engineering using disposable technology for enhanced production of hCTLA4Ig in transgenic rice cell cultures

Abstract

Two kinds of disposable bioreactors, air-lift disposable bioreactors (ADB) and wave disposable bioreactors (WDB) were compared with stirred-tank reactors (5-L STR). These bioreactors were successfully applied to transgenic rice cell cultures for the production of recombinant human cytotoxic T-lymphocyte antigen 4-immunoglobulin (hCTLA4Ig). In both systems, a fed-batch culture method was used to produce hCTLA4Ig efficiently by feeding concentrated amino acids and production levels were enhanced when dissolved oxygen (DO) level was regulated at 30% using pure oxygen sparging. Agitation and aeration rate during cultivation in ADB and WDB were determined by the same mixing time. The results in both disposable bioreactors showed similar values in maximum cell density (11.9 gDCW/L and 12.6 gDCW/L), doubling time (4.8- and 5.0-day), and maximum hCTLA4Ig concentration (43.7 and 43.3 mg/L). Relatively higher cell viability was sustained in the ADB whereas hCTLA4Ig productivity was 1.2-fold higher than that in WDB. The productivity was improved by increasing aeration rate (0.2 vvm). Overall, our experiments demonstrate pneumatically driven disposable bioreactors are applicable for the production of recombinant proteins in plant cell cultures. These results will be useful for development and scale-up studies of disposable bioreactor systems for transgenic plant cell cultures.

Efficient increase of the novel recombinant human plasminogen activator expression level and stability through the use of homozygote transgenic rabbits

“…The rhPA protein is shown as two bands with molecular weight of 39 and 37 kda in SdS-PAGE gels (Fig. 7), which is in accordance with the observations of a previous study (38). Lane 8 represents the protein Marker (SN123, Nanjing Shengxing Biology company, Nanjing, china), Lanes 2 and 5 are reteplase National Institute for Biological Standards and control, 20 and 1 µg, respectively.…”

Rapid expansion in the WAVE bioreactor of clinical scale cells for tumor immunotherapy

“…[38][39][40] In order to rapidly expand these T cells under GMP conditions for clinical trials in the future, the WAVE bioreactor, according to our analysis, might be a preferred method of cell expansion for some cell subsets or phenotypes. WAVE Bioreactors are attractive for wide range of specialized applications, including production of clinical-grade viral vectors, 41 human mesenchymal stem cells, 26 plant cell culture, 42 and culture of malarial parasites. 25,43 However, they have some disadvantages, such as, extra cost of disposable cellbags, size limit of 500 L, and lack of distant-control or monitoring system.…”

N-glycan Remodeling Using Mannosidase Inhibitors to Increase High-mannose Glycans on Acid α-Glucosidase in Transgenic Rice Cell Cultures

“…Cell mass was estimated by measuring dry cell weight. Assay of relative cell viability was performed through measurement of absorbance at 485 nm using triphenyl tetrazolium chloride solution 21 .…”