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Biotechnol Prog ; 30(6): 1457-68, 2014.
Article in English | MEDLINE | ID: mdl-25079388

ABSTRACT

A unique and nontraditional approach using glutamine and asparagine supplements for CHO-glutamine synthetase (GS) cell lines was studied. In our experiments, we found that a decrease in pH and an increase in cell death occurred in production phase of a GS cell line, leading to reduced antibody expression and lower antibody yields. The experimental results and the statistical analysis (ANOVA) indicated that additions of glutamine and asparagine in the basal and feed media were effective to buffer the cell culture pH, reduce lactate generation, maintain a higher cell viability profile, and improve antibody productivity. In bench-top bioreactors, glutamine and asparagine supplementation helped to prevent cell death, improve antibody yield, and reduce base usage. Glutamine is normally excluded from culture media for GS cell lines to prevent the bypass of selection pressure. In this study, however, the addition of glutamine did not affect cell population homogeneity, protein quality, or decrease antibody yield of two GS cell lines.


Subject(s)
Antibodies/metabolism , Asparagine/metabolism , Culture Media/chemistry , Glutamine/metabolism , Recombinant Proteins/metabolism , Animals , Asparagine/pharmacology , Bioreactors , CHO Cells , Cell Culture Techniques , Cell Survival/drug effects , Cricetinae , Cricetulus , Culture Media/metabolism , Culture Media/pharmacology , Flow Cytometry , Glutamate-Ammonia Ligase , Glutamine/pharmacology
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