Cell retention–chemostat studies of hybridoma cells—analysis of hybridoma growth and metabolism in continuous suspension culture in serum‐free medium

Hiller, Gregory; Clark, Douglas S.; Blanch, Harvey W.

doi:10.1002/bit.260420206

Cited by 86 publications

(73 citation statements)

References 26 publications

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“…This computational method is quite reliable because the experimental error is not amplified in the calculation. On the other hand, constant and variable specific production rates have been reported by other investigators (Chang et al, 1995;Hiller et al, 1993;Linardos et al, 1991). In some cases, the change in specific production rate might be caused by a short culture span and experimental errors.…”

Section: Communication To the Editor Discussionmentioning

confidence: 88%

High cell density and high monoclonal antibody production through medium design and rational control in a bioreactor

Xie¹,

Wang²

2000

Biotechnol. Bioeng.

102

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A simple feeding strategy was developed and successfully employed for nutritional control in a 2‐L fed‐batch culture of hybridoma cells. A previously developed stoichiometric model for animal cell growth was used to design a supplemental medium for feeding. Undialyzed fetal bovine serum and trace metals (Fe2+, SeO 32−, Li+, Zn2+, and Cu2+) were fed to the cells periodically in addition to the automatic feeding of other nutrients in the supplemental medium. In this study, the maximum viable cell density was increased from 6.3 × 106 to 1.7 × 107 cells/mL, and the culture span was extended from 340 to 550 hours. The final monoclonal antibody titer achieved was 2400 mg/L. The specific production rates for ammonia and lactate were further reduced from 0.0045 and 0.0048 in our previous fed‐batch experiments to 0.0028 and 0.0036 mmol/109 cell h, respectively. Only 3.4% of the total glucose consumption was converted into lactate, compared to 67% in a conventional batch culture.

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Section: Communication To the Editor Discussionmentioning

confidence: 88%

High cell density and high monoclonal antibody production through medium design and rational control in a bioreactor

Xie¹,

Wang²

2000

Biotechnol. Bioeng.

102

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“…These results are similar to those reported by Hiller et al for this cell line at low growth rates. 32 The production rate of ammonia, 0.032 mmol/109 cells/h (see Table I), was only 70% of the consumption rate of glutamine, indicating that X-D hybridomas may have significant glutamine amidotransferase activity.…”

Section: Results and Discussion L-13c Glucose Infusionmentioning

confidence: 98%

Examination of primary metabolic pathways in a murine hybridoma with carbon‐13 nuclear magnetic resonance spectroscopy

Mancuso

Sharfstein

Tucker

et al. 1994

Biotech & Bioengineering

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Primary metabolism of a murine hybridoma was probed with (13)C nuclear magnetic resonance (NMR) spectroscopy. Cells cultured in a hollow fiber bioreactor were serially infused with [1-(13)C] glucose, [2-(13)C] glucose, and [3-(13)C] glutamine. In vivo spectroscopy of the culture was used in conjunction with off-line spectroscopy of the medium to determine the intracellular concentration of several metabolic intermediates and to determine fluxes for primary metabolic pathways. Intracellular concentrations of pyruvate and alanine were very high relative to levels observed in normal quiescent mammalian cells. Estimates made from labeling patterns in lactate indicate that 76% of pyruvate is derived directly from glycolysis; some is also derived from the malate shunt, the pyruvate/melate shuttle associated with lipid synthesis and the pentose phosphate pathway. The rate of formation of pyruvate from the pentose phosphate pathway was estimated to be 4% of that from glycolysis; This value is a lower limit and the actual value may be higher. Incorporation of pyruvate into the tricarboxylic acid (TCA) cycle appears to occur through only pyruvate dehydrogenase; no pyruvate carboxylase activity was detected. The malate shunt rate was approximately equal to the rate of glutamine uptake. The rate of incorporation of glucosederived acetyl-CoA into lipids was 4% of the glucose uptake rate. The TCA cycle rate between isocitrate and alpha-ketoglutarate was 110% of the glutamine uptake rate.

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“…High molecular weight products were reported to be retained by the membranes, inducing problems of product stability (Hiller et al, 1993). In several studies, cell bleeding was used to limit cell density (Buntenmeyer et al, 1994;Heath, 1995, 1996).…”

Section: Hollow Fibermentioning

confidence: 99%

“…Commercial modules (cellulose-ester fibers, 500-600 cm 2 , Microgon, Laguna Hills, CA, USA) were used by several investigators as retention devices for suspended cultures, either externally (Hiller et al, 1993;Zhang et al, 1993) or internally (Kyung et al, 1994), or even as microbioreactor for immobilized cultures (Tanase et al, 1997). The largest module available to date by Microgon for perfusion culture has a surface of 3.3 m 2 and is applicable to bioreactors ranging from 90-250 L. Another company proposes an original technology of Alternating Tangential Flow (ATF, Refine Technology Co., Edison, NJ, USA).…”

Section: Hollow Fibermentioning

confidence: 99%

Potential of cell retention techniques for large‐scale high‐density perfusion culture of suspended mammalian cells

Voisard

Meuwly

Ruffieux

et al. 2003

Biotech & Bioengineering

245

150

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This review focuses on cultivation of mammalian cells in a suspended perfusion mode. The major technological limitation in the scaling-up of these systems is the need for robust retention devices to enable perfusion of medium as needed. For this, cell retention techniques available to date are presented, namely, cross-flow filters, hollow fibers, controlled-shear filters, vortex-flow filters, spin-filters, gravity settlers, centrifuges, acoustic settlers, and hydrocyclones. These retention techniques are compared and evaluated for their respective advantages and potential for large-scale utilization in the context of industrial manufacturing processes. This analysis shows certain techniques have a limited range of perfusion rate where they can be implemented (most microfiltration techniques). On the other hand, techniques were identified that have shown high perfusion capacity (centrifuges and spin-filters), or have a good potential for scale-up (acoustic settlers and inclined settlers). The literature clearly shows that reasonable solutions exist to develop large-scale perfusion processes.

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Cell retention–chemostat studies of hybridoma cells—analysis of hybridoma growth and metabolism in continuous suspension culture in serum‐free medium

Cited by 86 publications

References 26 publications

High cell density and high monoclonal antibody production through medium design and rational control in a bioreactor

High cell density and high monoclonal antibody production through medium design and rational control in a bioreactor

Examination of primary metabolic pathways in a murine hybridoma with carbon‐13 nuclear magnetic resonance spectroscopy

Potential of cell retention techniques for large‐scale high‐density perfusion culture of suspended mammalian cells

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