Amino acid consumption in naïve and recombinant CHO cell cultures: producers of a monoclonal antibody

Потребление аминокислот в наивных и рекомбинантных культурах клеток CHO — продуцентах моноклонального антитела
Leydi Maribel Carrillo‐Cocom, T. Genel-Rey, Diana Araíz-Hernández, Felipe López‐Pacheco, Julián López-Meza, María del Refugio Rocha-Pizaña, Alicia Ramírez‐Medrano, Mario Moisés Álvarez
2014-05-05

CHO cell culturesamino acid consumptionamino acid supplementationfed-batch cultivationmonoclonal antibody production
Most commercial media for mammalian cell culture are designed to satisfy the amino acid requirements for cell growth, but not necessarily those for recombinant protein production. In this study, we analyze the amino acid consumption pattern in naïve and recombinant Chinese hamster ovary (CHO) cell cultures. The recombinant model we chose was a CHO-S cell line engineered to produce a monoclonal antibody. We report the cell concentration, product concentration, and amino acid concentration profiles in naïve and recombinant cell cultures growing in CD OptiCHO™ medium with or without amino acid supplementation with a commercial supplement (CHO CD EfficientFeed™ B). We quantify and discuss the amino acid demands due to cell growth and recombinant protein production during long term fed batch cultivation protocols. We confirmed that a group of five amino acids, constituting the highest mass fraction of the product, shows the highest depletion rates and could become limiting for product expression. In our experiments, alanine, a non-important mass constituent of the product, is in high demand during recombinant protein production. Evaluation of specific amino acid demands could be of great help in the design of feeding/supplementation strategies for recombinant mammalian cell cultures.
1
Alanine showed high demand during recombinant protein production despite contributing little to the antibody’s mass composition.
2
Amino acid demand was quantified separately for cell growth and recombinant monoclonal antibody production using CD OptiCHO™ medium with or without CHO CD EfficientFeed™ B supplementation.
3
Five amino acids comprising the antibody’s highest mass fractions exhibited the greatest depletion rates and could limit recombinant product expression.
4
Specific amino acid demand measurements can guide feeding and supplementation strategies for recombinant mammalian cell cultures.
5
The study characterizes amino acid consumption in naïve and monoclonal-antibody-producing recombinant CHO-S cell cultures during long-term fed-batch cultivation.

Naïve and recombinant Chinese hamster ovary (CHO) cell cultures, including a CHO-S cell line producing a monoclonal antibody

Amino acid consumption and specific demands associated with cell growth and recombinant monoclonal-antibody production, including depletion and potential nutrient limitation

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2014-05-05
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Leydi Maribel Carrillo‐Cocom
T. Genel-Rey
Diana Araíz-Hernández
Felipe López‐Pacheco
Julián López-Meza
María del Refugio Rocha-Pizaña
Alicia Ramírez‐Medrano
Mario Moisés Álvarez
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