Increased CHO cell fed-batch monoclonal antibody production using the autophagy inhibitor 3-MA or gradually increasing osmolality

Nasseri, S. Soroush; Ghaffari, Navid; Braasch, Katrin; Jardon, Mario A.; Butler, Michael; Kennard, Malcolm L.; Gopaluni, R. Bhushan; Piret, James M.

doi:10.1016/j.bej.2014.06.027

Cited by 22 publications

(22 citation statements)

References 38 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…ChK2 437.89.56 referred to as CHO‐K1SV, from Pfizer Inc., derived from Lonza's CHO‐K1SV cell line producing human monoclonal anti‐interleukin 1 β IgG1 antibody . The CHO‐S cell line producing a human IgG1 was from Australian Institute for Bioengineering and Nanotechnology …”

Section: Methodsmentioning

confidence: 99%

Effects of cysteine, asparagine, or glutamine limitations in Chinese hamster ovary cell batch and fed‐batch cultures

Ghaffari

Jardon

Krahn

et al. 2019

Biotechnology Progress

Self Cite

View full text Add to dashboard Cite

Amino acid availability is a key factor that can be controlled to optimize the productivity of fed‐batch cultures. To study amino acid limitation effects, a serum‐free chemically defined basal medium was formulated to exclude the amino acids that became depleted in batch culture. The effect of limiting glutamine, asparagine, and cysteine on the cell growth, metabolism, antibody productivity, and product glycosylation was investigated in three Chinese hamster ovary (CHO) cell lines (CHO‐DXB11, CHO‐K1SV, and CHO‐S). Cysteine limitation was detrimental to both cell proliferation and productivity for all three CHO cell lines. Glutamine limitation reduced growth but not cell specific productivity, whereas asparagine limitation had no significant effect on either growth or cell specific productivity. Neither glutamine nor asparagine limitation significantly affected antibody glycosylation. Replenishing the CHO‐DXB11 culture with cysteine after 1 day of cysteine limitation allowed the cells to partially recover their growth and productivity. This recovery was not observed after 2 days of cysteine limitation. Based on these findings, a fed‐batch protocol was developed using single or mixed amino acid supplementation. Although cell density and antibody concentration were lower compared to a commercial feed, the feeds based on cysteine supplementation yielded comparable cell specific productivity. Overall, this study showed that different amino acid limitations have varied effects on the performance of CHO cell cultures and that maintaining cysteine availability is a critical process parameter for the three cell lines investigated.

show abstract

Section: Methodsmentioning

confidence: 99%

Effects of cysteine, asparagine, or glutamine limitations in Chinese hamster ovary cell batch and fed‐batch cultures

Ghaffari

Jardon

Krahn

et al. 2019

Biotechnology Progress

Self Cite

View full text Add to dashboard Cite

show abstract

“…For CHO cells, specific cell growth rate has been reported to decrease linearly with increasing expansion medium osmolality between 316–450 mOsm/kg . However, gradual increase of osmolality up to ~450 mOsm/kg has also been shown to significantly increase specific productivity and mAb production in a CHO cell process . Therefore, when designing cell culture media, the final osmolality should be taken into consideration in relation to the overall salt and bulk ion supply.…”

Section: Key Components Of Mammalian Cell Culture Mediamentioning

confidence: 99%

Cell culture media for recombinant protein expression in Chinese hamster ovary (CHO) cells: History, key components, and optimization strategies

Ritacco

Khetan

2018

Biotechnology Progress

182

138

View full text Add to dashboard Cite

The culture of Chinese Hamster Ovary (CHO) cells for modern industrial applications, such as expression of recombinant proteins, requires media that support growth and production. Such media must support high viable cell densities while also stimulating the synthesis and extracellular transport of biologic products. Early media development efforts in this area yielded basic formulations to sustain growth, viability, and cellular function, albeit comprising animal sourced components, and complex constituents used in batch culture mode. Subsequent improvements included the development of serum‐free and chemically defined (CD) media, the identification of critical nutrients, growth factors, and potentially inhibitory or toxic cellular metabolites, and the use of fed‐batch and perfusion culture techniques to optimize nutrient delivery while minimizing accumulation of unwanted waste products. This review is comprised of sections covering milestones in the evolution of mammalian cell culture media, nutrient composition and formulation requirements, optimization strategies, consistency and scalability of powder and liquid media preparation for industrial applications, and key recent advances driving progress in CHO cell culture medium design and development. © 2018 American Institute of Chemical Engineers Biotechnol. Prog., 34:1407–1426, 2018

show abstract

“…A particularly successful example was the transfection of two inducible anti‐apoptotic genes into Chinese Hamster Ovary (CHO) cells that enabled increased fed‐batch viable cell concentrations as well as recombinant protein production (Figueroa et al, ). More recently, factors that modulate autophagy have also been used to increase CHO cell protein production up to ∼3‐fold in fed‐batch, in particular using 3‐methyladenine (a putative inhibitor that can also induce autophagy with prolonged exposure) (Baek, Kim, Kim, Lee, & Lee, ; Jardon et al, ; Nasseri et al, ). Combined strategies to inhibit apoptosis and induce autophagy have also been developed (Lee, Ha, Park, & Lee, ).…”

Section: Introductionmentioning

confidence: 99%

Types of cell death and apoptotic stages in Chinese Hamster Ovary cells distinguished by Raman spectroscopy

Rangan

Kamal

Konorov

et al. 2017

Biotech & Bioengineering

Self Cite

View full text Add to dashboard Cite

Cell death is the ultimate cause of productivity loss in bioreactors that are used to produce therapeutic proteins. We investigated the ability of Raman spectroscopy to detect the onset and types of cell death for Chinese Hamster Ovary (CHO) cells-the most widely used cell type for therapeutic protein production. Raman spectroscopy was used to compare apoptotic, necrotic, autophagic, and control CHO cells. Several specific nucleic acid-, protein-, and lipid-associated marker bands within the 650-850 cm spectral region were identified that distinguished among cells undergoing different modes of cell death; supporting evidence was provided by principal component analysis (PCA) of the full spectral data. In addition to comparing the different modes of cell death, normal cells were compared to cells sorted at several stages of apoptosis, in order to explore the potential for early detection of apoptosis. Different stages of apoptosis could be distinguished via Raman spectroscopy, with multiple changes observed in nucleic acid peaks at early stages whereas an increase in lipid signals was a feature of late apoptosis/secondary necrosis.

show abstract

Increased CHO cell fed-batch monoclonal antibody production using the autophagy inhibitor 3-MA or gradually increasing osmolality

Cited by 22 publications

References 38 publications

Effects of cysteine, asparagine, or glutamine limitations in Chinese hamster ovary cell batch and fed‐batch cultures

Effects of cysteine, asparagine, or glutamine limitations in Chinese hamster ovary cell batch and fed‐batch cultures

Cell culture media for recombinant protein expression in Chinese hamster ovary (CHO) cells: History, key components, and optimization strategies

Types of cell death and apoptotic stages in Chinese Hamster Ovary cells distinguished by Raman spectroscopy

Contact Info

Product

Resources

About