Growth, Metabolic, And Antibody Production Kinetics Of Hybridoma Cell Culture: 1. Analysis Of Data From Controlled Batch Reactors

Öztürk, Sadettin S.; Palsson, Bernhard Ø.

doi:10.1021/bp00012a001

Cited by 68 publications

(39 citation statements)

References 35 publications

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“…This different environment interacts with cell development, altering morphology and membrane permeability, as well as surface tension and osmotic pressure 31,35,39 . Maximum specific rates found in this work were slightly higher than lactate and ammonium specific production rates and glucose specific consumption rate reported by Ozturk and Palsson 40 (0.13, 0.002, and 0.14 h -1 , respectively). On the contrary, the maximum specific consumption rates for glutamine were very similar (≈ 0.03 h -1 ).…”

Section: -Evolution Of Antibody Concentration (Samples Taken From Thecontrasting

confidence: 72%

Diffusion and Inhibition Processes in a Hollow-fiber Membrane Bioreactor for Hybridoma Culture. Development of a Mathematical Model

Legazpi

Laca

Collado

et al. 2016

Chem.Biochem.Eng.Q.

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The performance of a hollow-fiber membrane bioreactor (HFBR) (molecular weight cut-off 30 kD, fiber surface area 2050 cm 2 ) containing a culture of hybridoma cells has been investigated. Experimental data were used as basis to develop a model of general application. Concentrations of fundamental nutrients (glucose and glutamine), inhibitory products (ammonium and lactate), and monoclonal antibodies (MAb) against bovine lactoferrin (IgG 1 ) were monitored over time. Exchange of nutrients and products occurred across the capillary surface, whereas cells and MAb remained in the extra-capillary space (ECS). A protein-free culture medium (Hybrimax) with and without antibiotics was used. In both cases, the final MAb concentration was the same; however, antibiotic presence slowed down the time to achieve this concentration. Diffusion assays have been carried out in order to support the development of a mathematical model that describes the performance of the HFBR, including mass transfer and reaction terms. Inhibition by ammonium and lactate has been considered in the kinetics, providing model results consistent with experimental data. Further research with other cell lines and/or culture media will allow to broaden the field of application of this model for general use in HFBR systems.

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Section: -Evolution Of Antibody Concentration (Samples Taken From Thecontrasting

confidence: 72%

Diffusion and Inhibition Processes in a Hollow-fiber Membrane Bioreactor for Hybridoma Culture. Development of a Mathematical Model

Legazpi

Laca

Collado

et al. 2016

Chem.Biochem.Eng.Q.

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“…The apparent growth rates ( app ) were calculated from the time profiles of viable cells plotted against the integral of viable cells. Death rates (k d ) were calculated from the time profiles of dead cells against the integral of viable cells (49). A ratio of app and k d for the induced cells relative to the noninduced cells was calculated.…”

Section: Construction Of Plasmidsmentioning

confidence: 99%

The Chaperone Function of hsp70 Is Required for Protection against Stress-Induced Apoptosis

Mosser

Caron

Bourget

et al. 2000

Molecular and Cellular Biology

649

479

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Cellular stress can trigger a process of self-destruction known as apoptosis. Cells can also respond to stress by adaptive changes that increase their ability to tolerate normally lethal conditions. Expression of the major heat-inducible protein hsp70 protects cells from heat-induced apoptosis. hsp70 has been reported to act in some situations upstream or downstream of caspase activation, and its protective effects have been said to be either dependent on or independent of its ability to inhibit JNK activation. Purified hsp70 has been shown to block procaspase processing in vitro but is unable to inhibit the activity of active caspase 3. Since some aspects of hsp70 function can occur in the absence of its chaperone activity, we examined whether hsp70 lacking its ATPase domain or the C-terminal EEVD sequence that is essential for peptide binding was required for the prevention of apoptosis. We generated stable cell lines with tetracycline-regulated expression of hsp70, hsc70, and chaperone-defective hsp70 mutants lacking the ATPase domain or the C-terminal EEVD sequence or containing AAAA in place of EEVD. Overexpression of hsp70 or hsc70 protected cells from heat shock-induced cell death by preventing the processing of procaspases 9 and 3. This required the chaperone function of hsp70 since hsp70 mutant proteins did not prevent procaspase processing or provide protection from apoptosis. JNK activation was inhibited by both hsp70 and hsc70 and by each of the hsp70 domain mutant proteins. The chaperoning activity of hsp70 is therefore not required for inhibition of JNK activation, and JNK inhibition was not sufficient for the prevention of apoptosis. Release of cytochrome c from mitochondria was inhibited in cells expressing full-length hsp70 but not in cells expressing the protein with ATPase deleted. Together with the recently identified ability of hsp70 to inhibit cytochrome c-mediated procaspase 9 processing in vitro, these data demonstrate that hsp70 can affect the apoptotic pathway at the levels of both cytochrome c release and initiator caspase activation and that the chaperone function of hsp70 is required for these effects.Protein-damaging stresses, such as exposure of cells to elevated temperatures, activate an adaptive response leading to the increased synthesis of a group of proteins that regulate protein-folding processes (reviewed in reference 43). Members of the hsp70 family of molecular chaperones recognize nonnative domains that are exposed during protein translation, membrane translocation, oligomerization, and ultimately degradation. The abundant cytoplasmic and nuclear protein hsc70 is assisted in this task by the highly inducible hsp70 protein, whose synthesis is controlled by the level of nonnative protein substrates. Conditions that alter protein structure can result in the exposure of hydrophobic regions that are normally buried within the molecule, leading to their aggregation and loss of function. The ability of hsp70 to compete for binding to these hydrophobic regions coupled with an ATP...

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“…We would like to acknowledge laboratory help by Cynthia Concannon and Mark Riley. This work was first presented at the Annual AIChE Meeting, San Francisco, November [5][6][7][8][9][10] 1989.…”

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confidence: 99%

Effect of medium osmolarity on hybridoma growth, metabolism, and antibody production

Öztürk

Palsson

1991

Biotech & Bioengineering

Self Cite

180

106

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Osmolarity is an important process variable during the cultivation of mammalian cells in vitro. Cell culture medium is designed to have osmolarity in the range of 260 and 320 milliosmoles (mOsm), basically to mimic the osmolarity of serum at 290 mOsm/kg. Fragmented information on the response of hybridoma cells to elevated osmolarity is available. A decrease in cell growth and an increase in antibody production were reported at high o~molarities.~~ Influence of osmolarity on antibody productivity seems to be cell line dependent.12 Here we report the effects of osmolarity on hybridoma cell growth, metabolism, and antibody synthesis. The osmolarity of the medium was altered by the addition of both ionic and non-ionic substances. MATERIALS AND METHODS Cell lines, Media, and Culture MaintenanceA murine hybridoma cell line, 167.465.3, was used in the experiments. This cell line was provided by Dr. Latham Claflin from the Medical Center at The University of Michigan. The antibody produced by this cell line is an kG1, directed against phosphorylcholine.2 Hybridoma :ells were made by fusion of BALB/c spleen cells with the nonsecreting plasmacytoma fusion line P3X63-488.653. Antibody was generated from mice immuiized with PC-keyhole limpet henemocyanin (KLH). "11s were propagated in T-flasks (Bellco Glass, Inc., Vineland, NJ) at 37°C in a humidified incubator under 5% COz. The media used was Iscove's Modified Dul-)ecco7s Medium (IMDM, Gibco Laboratories, Grand sland, NY) containing 5% Fetal Bovine Serum (FBS, 3ibco). The media was supplemented by 100 Units/mL )otassium penicillin G, and 100 pg/mL streptomycin ulfate (Sigma Chemical, St. Louis, MO). Effect of Osmolarity on Cell Physiology:ells were inoculated into IMDM medium at different jsmolarities. Osmolarity in IMDM medium was in-:reased by the addition of either an ionic (NaC1 or PBS) )r nonionic (sucrose) species. The IMDM medium was repared from powder and the pH was adjusted to 7.4 by the addition of sodium bicarbonate. Medium osmolarity was measured using an Osmometer (Osmette, model 2007, Precision Systems, Inc., Natick, MA) as 290 mOsm. Media with osmolarities of about twice this value (580 mOsm) were prepared by dissolving IMDM in (1) 9 g/L NaC1, (2) in PBS, and (3) in 0.31M sucrose solution. The pH was also adjusted to 7.4 in these media. The osmolarities in these media preparations were measured. Then these IMDM preparations were mixed with standard IMDM media at 290 mOsm to give final osmolarities of 290, 338, 386, 435, and 580 mOsm.Cells growing exponentially in ordinary IMDM with 5% FBS were spun down at 200g for 10 min. They were then washed with fresh IMDM and were inoculated with IMDM with different osmolarities. Fetal bovine serum was added to all the media preparations at 5%. Batch cultivations were carried out in 100-mL spinner flasks at an initial cell density of 4 x lo4 cells/mL. The volume in spinner flasks was 60 mL and they were kept at 37°C in a humidified incubator under 5% C02. Agitation at 100 rpm was provided by a MultiStir magnetic stir...

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Growth, Metabolic, And Antibody Production Kinetics Of Hybridoma Cell Culture: 1. Analysis Of Data From Controlled Batch Reactors

Cited by 68 publications

References 35 publications

Diffusion and Inhibition Processes in a Hollow-fiber Membrane Bioreactor for Hybridoma Culture. Development of a Mathematical Model

Diffusion and Inhibition Processes in a Hollow-fiber Membrane Bioreactor for Hybridoma Culture. Development of a Mathematical Model

The Chaperone Function of hsp70 Is Required for Protection against Stress-Induced Apoptosis

Effect of medium osmolarity on hybridoma growth, metabolism, and antibody production

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