Glutamine substitution: the role it can play to enhance therapeutic protein production

Ha, Tae Kwang; Lee, Gyun Min

doi:10.4155/pbp.15.6

Cited by 11 publications

(4 citation statements)

References 97 publications

(95 reference statements)

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…Glutamate, as the replacement of glutamine, plays an important role in cell growth and cell survival. It is a major nitrogen source and also a main energy source in CHO cell cultures [ 76 , 77 ]. Our result of increased q Glu with increase of glucose feed concentration was consistent with previous observations, where glutamate was used as a glutamine replacement [ 68 ].…”

Section: Discussionmentioning

confidence: 99%

High glucose and low specific cell growth but not mild hypothermia improve specific r-protein productivity in chemostat culture of CHO cells

et al. 2018

View full text Add to dashboard Cite

In the biopharmaceutical sector, Chinese hamster ovary (CHO) cells have become the host of choice to produce recombinant proteins (r-proteins) due to their capacity for correct protein folding, assembly, and posttranslational modification. However, the production of therapeutic r-proteins in CHO cells is expensive and presents insufficient production yields for certain proteins. Effective culture strategies to increase productivity (qp) include a high glucose concentration in the medium and mild hypothermia (28–34 °C), but these changes lead to a reduced specific growth rate. To study the individual and combined impacts of glucose concentration, specific growth rate and mild hypothermia on culture performance and cell metabolism, we analyzed chemostat cultures of recombinant human tissue plasminogen activator (rh-tPA)-producing CHO cell lines fed with three glucose concentrations in feeding media (20, 30 and 40 mM), at two dilution rates (0.01 and 0.018 1/h) and two temperatures (33 and 37 °C). The results indicated significant changes in cell growth, cell cycle distribution, metabolism, and rh-tPA productivity in response to the varying environmental culture conditions. High glucose feed led to constrained cell growth, increased specific rh-tPA productivity and a higher number of cells in the G2/M phase. Low specific growth rate and temperature (33 °C) reduced glucose consumption and lactate production rates. Our findings indicated that a reduced specific growth rate coupled with high feed glucose significantly improves r-protein productivity in CHO cells. We also observed that low temperature significantly reduced qp, but not cell growth when dilution rate was manipulated, regardless of the glucose concentration or dilution rate. In contrast, we determined that feed glucose concentration and consumption rate were the dominant aspects of the growth and productivity in CHO cells by using multivariate analysis.

show abstract

Section: Discussionmentioning

confidence: 99%

High glucose and low specific cell growth but not mild hypothermia improve specific r-protein productivity in chemostat culture of CHO cells

et al. 2018

View full text Add to dashboard Cite

show abstract

“…[16,21,[23][24][25][26][27] Academic and industrial research groups have spent over 30 years developing methods to minimize ammonia and waste product accumulation with better progress made reducing lactate than ammonia accumulation. [9,[28][29][30][31][32] Most commonly, exogenous ammonia is utilized to simulate ammonia stress to disconnect the metabolic effects. [6,7,12,33] Yet, the basic rules of thumb to minimize ammonia accumulation are to use alternative carbon and nitrogen sources and improve process control of the carbon/nitrogen concentrations.…”

Section: Introductionmentioning

confidence: 99%

Transient ammonia stress on Chinese hamster ovary (CHO) cells yield alterations to alanine metabolism and IgG glycosylation profiles

Synoground

McGraw

Elliott

et al. 2021

Biotechnology Journal

View full text Add to dashboard Cite

Background Ammonia concentrations typically increase during mammalian cell cultures, mainly due to glutamine and other amino acid consumption. An early ammonia stress indicator is a metabolic shift with respect to alanine. To determine the underlying mechanisms of this metabolic shift, a Chinese hamster ovary (CHO) cell line with two distinct ages (standard and young) was cultured in parallel fed‐batch bioreactors with 0 mM or 10 mM ammonia added at 12 h. Reduced viable cell densities were observed for the stressed cells, while viability was not significantly affected. The stressed cultures had higher alanine, lactate, and glutamate accumulation. Interestingly, the ammonia concentrations were similar by Day 8.5 for all cultures. We hypothesized the ammonia was converted to alanine as a coping mechanism. Interestingly, no significant differences were observed for metabolite profiles due to cell age. Glycosylation analysis showed the ammonia stress reduced galactosylation, sialylation, and fucosylation. Transcriptome analysis of the standard‐aged cultures indicated the ammonia stress had a limited impact on the transcriptome, where few of the significant changes were directly related metabolite or glycosylation reactions. These results indicate that mechanisms used to alleviate ammonia stress are most likely controlled post‐transcriptionally, and this is where future research should focus.

show abstract

“…Aunque el medio de cultivo empleado en las fermentaciones contiene una gran cantidad de componentes entre aminoácidos, fuentes de carbono, sales minerales y otros, los únicos componentes del medio de cultivo que incluye el modelo cinético de Kontoravdi, (2007) son la glucosa y la glutamina. Estos son consumidos rápidamente por las células, y el metabolismo de ambos genera productos inhibidores del crecimiento y la síntesis de productos como el lactato y el amonio (Jang & Barford, 2000;Kwang & Lee, 2015;Ozturk, 2006).…”

Section: Ecuaciones Asociadas Al Crecimiento Celularunclassified

Heurística Y Simulación Computacional Para El Diseño De Un Sistema De Cultivo Celular

Vargas

Salazar

2023

HYW

View full text Add to dashboard Cite

Un sistema fermentativo en etapa de desarrollo tecnológico va dirigido a la producción del ingrediente farmacéuticamente activo (IFA) de una vacuna contra la Peste Porcina Clásica empleando la línea celular HEK293. El objetivo fue proponer una estrategia de optimización aplicada al diseño del sistema de cultivo celular empleando la heurística sobre la base de la analogía entre el sistema experimental real y la simulación computacional. Para realizar las simulaciones del proceso se seleccionó el modelo cinético (Kontoravdi et al., 2007). Se mostró que los sistemas de fermentación reales han sido operados en condiciones muy alejadas de las óptimas y que el comportamiento de estos responde al errado régimen de alimentación al biorreactor que provoca la acumulación de productos tóxicos del metabolismo y el apagado del biorreactor. A partir de estos resultados se propone una estrategia de optimización que tiene en cuenta las características específicas del sistema de cultivo celular, la complejidad del modelo cinético seleccionado y que aprovecha las prestaciones del software MATLAB.

show abstract

Glutamine substitution: the role it can play to enhance therapeutic protein production

Cited by 11 publications

References 97 publications

High glucose and low specific cell growth but not mild hypothermia improve specific r-protein productivity in chemostat culture of CHO cells

High glucose and low specific cell growth but not mild hypothermia improve specific r-protein productivity in chemostat culture of CHO cells

Transient ammonia stress on Chinese hamster ovary (CHO) cells yield alterations to alanine metabolism and IgG glycosylation profiles

Heurística Y Simulación Computacional Para El Diseño De Un Sistema De Cultivo Celular

Contact Info

Product

Resources

About