Glutamine replenishment and ammonia removal in hybridoma cell cultures via immobilized glutamine synthetase

Persistent Link:
http://hdl.handle.net/10150/278706
Title:
Glutamine replenishment and ammonia removal in hybridoma cell cultures via immobilized glutamine synthetase
Author:
Okeson, Carl D.
Issue Date:
1999
Publisher:
The University of Arizona.
Rights:
Copyright © is held by the author. Digital access to this material is made possible by the University Libraries, University of Arizona. Further transmission, reproduction or presentation (such as public display or performance) of protected items is prohibited except with permission of the author.
Abstract:
Hybridoma cells utilize glutamine as their primary nitrogen source and excrete ammonia as a metabolic waste product. This ammonia can quickly accumulate to toxic levels in hybridoma culture media, and can severely reduce monoclonal antibody production (Ozturk et al., 1991). The enzyme glutamine synthetase (E.C. 6.3.1.2), which catalyzes the reaction UNFORMATTED EQUATION FOLLOWS: NH₄⁺ + L-glutamate + ATP Mg²⁺ → L-glutamine + ADP +Pᵢ +H⁺, UNFORMATTED EQUATION ENDS was evaluated as a means of reducing ammonia and replenishing glutamine in hybridoma culture medium. The effect of each enzyme reactant on soluble glutamine synthetase activity was quantified, and enzymatic reaction equilibrium evaluated. Enzyme reaction rates in two culture media, both with and without serum, were compared. Glutamine synthetase was immobilized via three different methods, and their effects compared. Cell sensitivity to each enzymatic reactant was studied. Finally, immobilized glutamine synthetase was incorporated in a hybridoma cultivation, and its effect on culture characteristics evaluated.
Type:
text; Thesis-Reproduction (electronic)
Keywords:
Chemistry, Biochemistry.; Chemistry, Biochemistry.
Degree Name:
M.S.
Degree Level:
masters
Degree Program:
Graduate College; Agriculture and Biosystems Engineering
Degree Grantor:
University of Arizona

Full metadata record

DC FieldValue Language
dc.language.isoen_USen_US
dc.titleGlutamine replenishment and ammonia removal in hybridoma cell cultures via immobilized glutamine synthetaseen_US
dc.creatorOkeson, Carl D.en_US
dc.contributor.authorOkeson, Carl D.en_US
dc.date.issued1999en_US
dc.publisherThe University of Arizona.en_US
dc.rightsCopyright © is held by the author. Digital access to this material is made possible by the University Libraries, University of Arizona. Further transmission, reproduction or presentation (such as public display or performance) of protected items is prohibited except with permission of the author.en_US
dc.description.abstractHybridoma cells utilize glutamine as their primary nitrogen source and excrete ammonia as a metabolic waste product. This ammonia can quickly accumulate to toxic levels in hybridoma culture media, and can severely reduce monoclonal antibody production (Ozturk et al., 1991). The enzyme glutamine synthetase (E.C. 6.3.1.2), which catalyzes the reaction UNFORMATTED EQUATION FOLLOWS: NH₄⁺ + L-glutamate + ATP Mg²⁺ → L-glutamine + ADP +Pᵢ +H⁺, UNFORMATTED EQUATION ENDS was evaluated as a means of reducing ammonia and replenishing glutamine in hybridoma culture medium. The effect of each enzyme reactant on soluble glutamine synthetase activity was quantified, and enzymatic reaction equilibrium evaluated. Enzyme reaction rates in two culture media, both with and without serum, were compared. Glutamine synthetase was immobilized via three different methods, and their effects compared. Cell sensitivity to each enzymatic reactant was studied. Finally, immobilized glutamine synthetase was incorporated in a hybridoma cultivation, and its effect on culture characteristics evaluated.en_US
dc.typetexten_US
dc.typeThesis-Reproduction (electronic)en_US
dc.subjectChemistry, Biochemistry.en_US
dc.subjectChemistry, Biochemistry.en_US
thesis.degree.nameM.S.en_US
thesis.degree.levelmastersen_US
thesis.degree.disciplineGraduate Collegeen_US
thesis.degree.disciplineAgriculture and Biosystems Engineeringen_US
thesis.degree.grantorUniversity of Arizonaen_US
dc.identifier.proquest1396517en_US
dc.identifier.bibrecord.b39916881en_US
All Items in UA Campus Repository are protected by copyright, with all rights reserved, unless otherwise indicated.