Analysis of cellular metabolism of hybridoma cells at distinct physiological states

Gambhir, Anshu; Korke, Rashmi; Lee, Jong-Chan; Fu, Peng; Europa, Anna F.; Hu, Wei Shou

doi:10.1016/s1389-1723(03)80062-2

Cited by 95 publications

(57 citation statements)

References 30 publications

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“…Zhou et al [49] reported that cellular metabolism was improved in fed-batch cultures when glucose feeding was controlled by on-line measurement of oxygen-uptake rate. The yield of lactate on glucose (ΔL/ΔG) decreased from 1.16 to 0.16 mol/mol, which dramatically reduced by-product accumulation and improved process performance [47,50]. Similar metabolic shifts were investigated at the transcriptional and translational levels [51,52].…”

Section: Glycolysis and Lactate Productionmentioning

confidence: 91%

Towards dynamic metabolic flux analysis in CHO cell cultures

Ahn

Antoniewicz

2011

Biotechnology Journal

111

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Chinese hamster ovary (CHO) cells are the most widely used mammalian cell line for biopharmaceutical production, with a total global market approaching $100 billion per year. In the pharmaceutical industry CHO cells are grown in fed-batch culture, where cellular metabolism is characterized by high glucose and glutamine uptake rates combined with high rates of ammonium and lactate secretion. The metabolism of CHO cells changes dramatically during a fed-batch culture as the cells adapt to a changing environment and transition from exponential growth phase to stationary phase. Thus far, it has been challenging to study metabolic flux dynamics in CHO cell cultures using conventional metabolic flux analysis techniques that were developed for systems at metabolic steady state. In this paper we review progress on flux analysis in CHO cells and techniques for dynamic metabolic flux analysis. Application of these new tools may allow identification of intracellular metabolic bottlenecks at specific stages in CHO cell cultures and eventually lead to novel strategies for improving CHO cell metabolism and optimizing biopharmaceutical process performance.

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Section: Glycolysis and Lactate Productionmentioning

confidence: 91%

Towards dynamic metabolic flux analysis in CHO cell cultures

Ahn

Antoniewicz

2011

Biotechnology Journal

111

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“…To better control cell metabolism and avoid using of animal-derived raw materials and ill-defined hydrolysates, much progress has been made in understanding cell nutrient requirements and development of chemically defined medium including feed formulations over the last 10 years (Wurm 2004;Franek et al 2000;Huang et al 2010;Young 2013). More recently, metabolic flux analysis (MFA) has become a useful tool in medium development for hybridoma Europa et al 2000;Gambhir et al 2003) and CHO cell cultures (Nyberg et al 1999;Altamirano et al 2006), which can help quantifying nutrient demands and metabolite production in cell culture (Zhao et al 2009). …”

Section: Introductionmentioning

confidence: 99%

Responses of CHO-DHFR cells to ratio of asparagine to glutamine in feed media: cell growth, antibody production, metabolic waste, glutamate, and energy metabolism

Zhang

Wang

et al. 2016

Bioresour. Bioprocess.

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Background: Optimization of chemically defined medium has been a critical way to produce monoclonal antibody. Usually, amount of glutamine was added into the feed medium, but a half of asparagine was added. Our study found that asparagine was important in the antibody production phase. Increasing the ratio of asparagine to glutamine in feed medium for enhancement of antibody production in CHO-DHFR cell culture would be an efficient way. Results:We optimized the total amount and the ratio of the two vital amino acids in feed medium to increase antibody production. In this work, we have demonstrated that feeding medium of high ratio between asparagine and glutamine (FB-H) can enhance the cell density after reaching the stationary phase. Moreover, FB-H was shown to improve cell maintenance, and increased the antibody production. The metabolic flux analysis proved that ratio of asparagine to glutamine had little influence on glycolysis. Furthermore, the TCA cycle of FB-H was enhanced by 20 % compared to that of low ratio of asparagine to glutamine (FB-L). And the energy metabolism of FB-H was 22.6 % higher than that of FB-L. For the later, lactate can be less produced in FB-H. Conclusions:We should improve the ratio between asparagine and glutamine in feed medium properly under the premise of no influence on cell growth to achieve high mAb producing goal.

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“…In particular, animal cell cultures have received considerable attention (Bonarius, 1996;Nyberg, 1999;Gambhir, 2003). Recently, there has been an increasing interest on the application of metabolic flux analysis to plant cell culture (Schwender, 2004;Ratcliffe, 2006).…”

Section: Applications Of Metabolic Flux Analysismentioning

confidence: 99%

“…Since all the constraints are linear equalities, the analytic solution of this minimisation problem can be obtained, resulting in the expressions to estimate v u and v m that are typically seen in literature, (Stephanopoulos, 1998, Gambhir, 2003. Details about TMFA calculations can be found in chapter II, section 2.8.…”

Section: Preliminaries: Metabolic Flux Analysismentioning

confidence: 99%

Interval and Possibilistic Methods for Constraint-Based Metabolic Models

Estrada¹

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In the following site the reader will find updated information regarding this thesis. This includes corrections and clarifications, connections with future works, publications, software and tools, etc.http://science.ensilicio.com/Thesis/Thesis.html AgradecimientosLa tesis doctoral es un viaje emocionante, exigente y no exento de riesgos. Por suerte, resulta mucho mejor, más sencillo y más divertido, cuando uno tiene una lista de agradecimientos tan extensa como la mía.Estoy en deuda con mi director, Jesús Picó, por darme la oportunidad de trabajar en lo que me gusta, por descubrirme un tema fascinante y por dirigirme sabiamente en el desarrollo de esta tesis. También con mis supervisores durante mis estancias en el extranjero, Georges Bastin y Vassily Hatzimanikatis, porque me permitieron aprender de su trabajo, conocer sus instituciones y explorar dos países.Le agradezco a Antonio Sala que se acercase un buen día con una propuesta para colaborar (haciendo así fructíferas las reuniones en la biblioteca); y a Marta Tortajada que fuese mi coautora en varias ocasiones y que sea una de las personas cuyas ideas más me han ayudado. He aprendido mil cosas de otras personas -profesores, compañeros, colegas, revisores, etc.-y a todos les estoy agradecido.A los compañeros que encontré camino al doctorado les agradezco, sobretodo, que hicieran mi trabajo divertido. Agradezco cada café y cada conversación. Mención especial merecen mis principales compañeros de viaje, Pepe y Sergio, por su camaradería en el trabajo y en la vida.Es probable que sin el apoyo de otras personas no hubiese terminado esta tesis... y es seguro que de haberlo hecho sería irrelevante. A mi familia -Juan, María, Juan-les agradezco que sean mi refugio, porque la existencia de ese «lugar» me permite el lujo de ser temerario. A mis amigos en la vecindad les agradezco su interés por las victorias de cada día; y tanto a ellos como al resto, les doy las gracias por creer incondicionalmente que lo que hacía, fuese lo que fuese, era digno de admirar. Mi deuda es especial con Lucia porque ella fue la recompensa en los días de trabajo y sacrificio.Por último, y por encima de todo, quiero agradecer a mis padres todo lo que hicieron y el modo preciso en que lo hicieron. Sé que eso excede el ámbito de esta tesis, pero este párrafo es un buen lugar para mostrarles mi gratitud. Gracias. AbstractThis thesis is devoted to the study and application of constraint-based metabolic models. The objective was to find simple ways to handle the difficulties that arise in practice due to uncertainty (knowledge is incomplete, there is a lack of measurable variables, and those available are imprecise). With this purpose, tools have been developed to model, analyse, estimate and predict the metabolic behaviour of cells.The document is structured in three parts. First, related literature is revised and summarised. This results in a unified perspective of several methodologies that use constraint-based representations of the cell metabolism. Three outstanding methods are discussed i...

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Analysis of cellular metabolism of hybridoma cells at distinct physiological states

Cited by 95 publications

References 30 publications

Towards dynamic metabolic flux analysis in CHO cell cultures

Towards dynamic metabolic flux analysis in CHO cell cultures

Responses of CHO-DHFR cells to ratio of asparagine to glutamine in feed media: cell growth, antibody production, metabolic waste, glutamate, and energy metabolism

Interval and Possibilistic Methods for Constraint-Based Metabolic Models

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