Dataset Information

Kinetic simulation of a centrifugal bioreactor for high population density hybridoma culture.

ABSTRACT: Demand for increasingly complex post-translationally modified proteins, such as monoclonal antibodies (mAbs), necessitates the use of mammalian hosts for production. The focus of this article is a continuous centrifugal bioreactor (CCBR) capable of increasing volumetric productivity for mAb production through high density hybridoma culture, exceeding 10(8) cells/mL. At these extreme densities, environmental conditions such as substrate and inhibitor concentrations rapidly change dramatically affecting the growth rate. The development of a kinetic model predicting glucose, mAb, lactate, and ammonium concentrations based on dilution rate and cell density is shown in this article. Additionally, it is found that pH affects both growth rate and viability, and a range of 6.9-7.4 is needed to maintain growth rate above 90% of the maximum. Modeling shows that operating an 11.4 mL CCBR inoculated with 2.0 x 10(7) cells/mL at a dilution rate of 1.3 h(-1), results in a predicted growth rate 82% of the maximum value. At the same dilution rate increasing density to 6.0 x 10(7) cells/mL decreases the predicted growth rate to 60% of the maximum; however, by increasing dilution rate to 6.1 h(-1) the growth rate can be increased to 86% of the maximum. Using the kinetic model developed in this research, the concentration of glucose, mAb, lactate, and ammonium are all predicted within 13% of experimental results. This model and an understanding of how RPM impacts cell retention serve as valuable tools for maintaining high density CCBR cultures, ensuring maximum growth associated mAb production rates.

SUBMITTER: Detzel CJ

PROVIDER: S-EPMC2796705 | biostudies-literature | 2009 Nov-Dec

REPOSITORIES: biostudies-literature

ACCESS DATA

Publications

Kinetic simulation of a centrifugal bioreactor for high population density hybridoma culture.

Detzel Christopher J CJ Mason Derek J DJ Davis William C WC van Wie Bernard J BJ

Biotechnology progress 20091101 6

Demand for increasingly complex post-translationally modified proteins, such as monoclonal antibodies (mAbs), necessitates the use of mammalian hosts for production. The focus of this article is a continuous centrifugal bioreactor (CCBR) capable of increasing volumetric productivity for mAb production through high density hybridoma culture, exceeding 10(8) cells/mL. At these extreme densities, environmental conditions such as substrate and inhibitor concentrations rapidly change dramatically aff ...[more]

PMID: 19806634

Similar Datasets

Project description:The rat pancreatic progenitor cell line B-13 is of interest for research on drug metabolism and toxicity since the cells trans-differentiate into functional hepatocyte-like cells (B-13/H) when treated with glucocorticoids. In this study we investigated the trans-differentiation and liver-specific functions of B-13/H cells in a three-dimensional (3D) multi-compartment bioreactor, which has already been successfully used for primary liver cell culture. Undifferentiated B-13 cells were inoculated into the bioreactor system and exposed to dexamethasone to promote hepatic trans-differentiation (B-13/HT). In a second approach, pre-differentiated B-13 cells were cultured in bioreactors for 15 days to evaluate the maintenance of liver-typical functions (B-13/HP). During trans-differentiation of B-13 cells into hepatocyte-like cells in the 3D bioreactor system (approach B-13/HT), an increase in glucose metabolism and in liver-specific functions (urea and albumin synthesis; cytochrome P450 [CYP] enzyme activity) was observed, whereas amylase - characteristic for exocrine pancreas and undifferentiated B-13 cells - decreased over time. In bioreactors with pre-differentiated cells (approach B-13/HP), the above liver-specific functions were maintained over the whole culture period. Results were confirmed by gene expression and protein analysis showing increased expression of carbamoyl-phosphate synthase 1 (CPS-1), albumin, CYP2E1, CYP2C11 and CYP3A1 with simultaneous loss of amylase. Immunohistochemical studies showed the formation of 3D structures with expression of liver-specific markers, including albumin, cytokeratin (CK) 18, CCAAT/enhancer-binding protein beta (CEBP-β), CYP2E1 and multidrug resistance protein 2 (MRP2). In conclusion, successful culture and trans-differentiation of B-13 cells in the 3D bioreactor was demonstrated. The requirement for only one hormone and simple culture conditions to generate liver-like cells makes this cell type useful for in vitro research using 3D high-density culture systems.

Dataset Information

Kinetic simulation of a centrifugal bioreactor for high population density hybridoma culture.

Publications

Kinetic simulation of a centrifugal bioreactor for high population density hybridoma culture.

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure

Tweets