Assessment of long-term cryopreservation for production of hCTLA4Ig in transgenic rice cell suspension cultures

Kwon, Jun-Young; Jeong, Sun-Hee; Choi, Ji-Won; Pak, Yun-Young; Kim, Dong-Il

doi:10.1016/j.enzmictec.2013.05.008

Cited by 8 publications

(4 citation statements)

References 23 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…목적단백질 생산유도 4일 차 이후에는 세포 내부의 hCTLA4Ig의 양이 점차 감소하였다. 이는 세포의 생존도 감소와 세포 용해로 인해 세포 내부의 hCTLA4Ig가 용출되었기 때문이다 [21]. 하지만 세포가 용해 되면 세포내의 프로테아제가 배지내로 용출되어 목적단백질 의 분해가 일어나기 때문에 적절한 시기에 목적단백질이 분 비되는 것이 단백질의 품질에 있어 매우 중요하다.…”

Section: Resultsunclassified

Enhanced Production of hCTLA4Ig through Increased Permeability in Transgenic Rice Cell Cultures

Choi¹,

Cheon²,

Kwon³

et al. 2016

KSBB Journal

Self Cite

View full text Add to dashboard Cite

In this system, rice cells were genetically modified to express human cytotoxic T-lymphocyte antigen 4-immunoglobulin (hCTLA4Ig) using RAmy3D promoter induced by sugar depletion. Even though the target protein fused with signal sequence peptide, plant cell wall can be a barrier against secretion of recombinant proteins. Therefore, hCTLA4Ig can be trapped inside cell wall or remained in intracellular space. In this study, to enhance the secretion of hCTLA4Ig from cytoplasm and cell walls into the medium, permeabilizing agents, such as dimethyl sulfoxide (DMSO), Triton X-100 and Tween 20, were applied in transgenic rice cell cultures. When 0.5% (v/v) of DMSO was added in sugar-free medium, intracellullar hCTLA4Ig was increased, on the other hand, the secreted extracellular hCTLA4Ig was lower than that of control. DMSO did not give permeable effects on transgenic rice cell cultures. And Triton X-100 was toxic to rice cells and also did not give enhancing permeability of cells. When 0.05% (v/v) Tween 20 was added in rice cell cultures, however, intracellular hCTLA4Ig was lower than that of control cultures. And the maximum 44.76 mg/L hCTLA4Ig was produced for 10 days after induction, which was 1.4-fold increase compared to that of control cultures. Especially, Tween 20 at 0.05% (v/v) showed the positive effect on the secretion of hCTLA4Ig though the decrease of intracellular hCTLA4Ig. Also, Tween 20 as a non-toxic surfactant did not affect the cell growth, cell viability and protease activity. In conclusion, secretion of hCTLA4Ig could be increased by enhancing permeability of cells regardless of the cell growth, cell viability and protease activity.

show abstract

Section: Resultsunclassified

Enhanced Production of hCTLA4Ig through Increased Permeability in Transgenic Rice Cell Cultures

Choi¹,

Cheon²,

Kwon³

et al. 2016

KSBB Journal

Self Cite

View full text Add to dashboard Cite

show abstract

“…Furthermore, when transgenic cell lines are cultivated and undergo periodic subculturing for extended periods, the production levels of heterologous proteins may decrease or even cease completely because of gene silencing or recombination events. To cope with this issue, the generation of cell banks is a viable approach 43 , 44 , 45 . Therefore, although no notable changes on the expression of VP2 have been observed during the long-term propagation of the SMC-1 line, the establishment of cryopreservation approaches for carrot cell cultures could allow the generation of a cell bank to ensure productivity in the long term.…”

Section: Discussionmentioning

confidence: 99%

Establishment of the Daucus carota SMC-1 Cell Suspension Line for Poliovirus Vaccine Development

Morales-Aguilar,

Bolaños-Martínez,

Maldonado

et al. 2023

Planta Med

View full text Add to dashboard Cite

The development of virus-free, oral vaccines against poliovirus capable of inducing mucosal protective immunity is needed to safely combat this pathogen. In the present study, a carrot cell line expressing the poliovirus VP2 antigen was established at the level of callus and cell suspensions, exploring the effects of culture media (MS and B5), supplementation with urea, phytoregulators (2,4-D : KIN), and light conditions (continuous light, photoperiod, and total darkness). The best callus growth was obtained on B5 medium supplemented with 2 mg/L of 2,4-D + 2 mg/L kinetin and 0.0136 g/L of urea and in continuous light conditions. Suspension cultures of the SMC-1 line in 250 mL Erlenmeyer flasks had a maximum growth of 16.07 ± 0.03 g/L DW on day 12 with a growth rate of µ=0.3/d and a doubling time of 2.3 days. In a 2 L airlift bioreactor, the biomass yield achieved was 25.6 ± 0.05 g/L DW at day 10 with a growth rate of µ= 0.58/d and doubling time of 1.38 d. Cell growth was 1.5 times higher in bioreactors than in shake flasks, highlighting that both systems resulted in the accumulation of VP2 throughout the time in culture. The maximum VP2 yield in flasks was 387.8 µg/g DW at day 21, while in the reactor it was 550.2 µg/g DW at day 18. In conclusion, bioreactor-based production of the VP2 protein by the SMC-1 suspension cell line offers a higher productivity when compared to flask cultures, offering a key perspective to produce low-cost vaccines against poliomyelitis.

show abstract

“…Thus, Protalix's process provides an additional reference point to justify the feasibility of the process described in this Total Equipment Costs ($ million) Seed Train Production Reactor Extraction Clarification UF/DF DEAE Hupresin UF/DF 2 Unlisted F I G U R E 6 Total equipment costs for a greenfield single-product facility producing rrBChE in two-stage semicontinuous and two-stage batch mode operation broken down by manufacturing section. DEAE, Diethylethanolamine; rrBChE, rice recombinant butyrylcholinesterase; UF/DF, ultrafiltration/diafiltration [Color figure can be viewed at wileyonlinelibrary.com] (Kwon et al, 2013;Mustafa, de Winter, van Iren, & Verpoorte, 2011), but there is no protocol that can be universally applied to all species (Santos, Abranches, Fischer, Sack, & Holland, 2016). There is also an ongoing literature debate as to the potential immunogenicity of plant glycan structures.…”

Section: Alternate Case 1: New Facilitymentioning

confidence: 99%

Technoeconomic analysis of semicontinuous bioreactor production of biopharmaceuticals in transgenic rice cell suspension cultures

Corbin

McNulty

Macharoen

et al. 2020

Biotech & Bioengineering

View full text Add to dashboard Cite

Biopharmaceutical protein production using transgenic plant cell bioreactor processes offers advantages over microbial and mammalian cell culture platforms in its ability to produce complex biologics with simple chemically defined media and reduced biosafety concerns. A disadvantage of plant cells from a traditional batch bioprocessing perspective is their slow growth rate which has motivated us to develop semicontinuous and/or perfusion processes. Although the economic benefits of plant cell culture bioprocesses are often mentioned in the literature, to our knowledge no rigorous technoeconomic models or analyses have been published. Here we present technoeconomic models in SuperPro Designer® for the large‐scale production of recombinant butyrylcholinesterase (BChE), a prophylactic/therapeutic bioscavenger against organophosphate nerve agent poisoning, in inducible transgenic rice cell suspension cultures. The base facility designed to produce 25 kg BChE per year utilizing two‐stage semicontinuous bioreactor operation manufactures a single 400 mg dose of BChE for $263. Semicontinuous operation scenarios result in 4–11% reduction over traditional two‐stage batch operation scenarios. In addition to providing a simulation tool that will be useful to the plant‐made pharmaceutical community, the model also provides a computational framework that can be used for other semicontinuous or batch bioreactor‐based processes.

show abstract

Assessment of long-term cryopreservation for production of hCTLA4Ig in transgenic rice cell suspension cultures

Cited by 8 publications

References 23 publications

Enhanced Production of hCTLA4Ig through Increased Permeability in Transgenic Rice Cell Cultures

Enhanced Production of hCTLA4Ig through Increased Permeability in Transgenic Rice Cell Cultures

Establishment of the Daucus carota SMC-1 Cell Suspension Line for Poliovirus Vaccine Development

Technoeconomic analysis of semicontinuous bioreactor production of biopharmaceuticals in transgenic rice cell suspension cultures

Contact Info

Product

Resources

About