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J Biotechnol ; 218: 53-63, 2016 Jan 20.
Artigo em Inglês | MEDLINE | ID: mdl-26654938

RESUMO

Industrial fed-batch cultivation of mammalian cells is used for the production of therapeutic proteins such as monoclonal antibodies. Besides medium ensuring initial growth, feeding is necessary to improve growth, viability and antibody production. Established processes include a slight acidic main feed and a separate alkaline feed containing l-tyrosine and l-cysteine. Since l-cysteine is not stable at neutral pH, a new derivative, S-sulfocysteine, was tested in neutral pH feeds. In small scale fed-batch processes, the S-sulfocysteine process yielded a comparable maximum viable cell density, prolonged viability and increased titer compared to the two feed system. Bioreactor experiments confirmed the increase in specific productivity. In depth characterization of the monoclonal antibody indicated no change in the glycosylation, or charge variant pattern whereas peptide mapping experiments were not able to detect any integration of the modified amino acid in the sequence of the monoclonal antibody. Finally, the mechanism of action of S-sulfocysteine was investigated, and results pointed out the anti-oxidative potential of the molecule, mediated through an increase in superoxide dismutase enzyme levels and in the total intracellular glutathione pool. Finally, we propose that the increase in specific productivity obtained in the S-sulfocysteine process results from the anti-oxidative properties of the molecule.


Assuntos
Antioxidantes/farmacologia , Técnicas de Cultura Celular por Lotes/métodos , Meios de Cultura , Cisteína/análogos & derivados , Aminoácidos/metabolismo , Animais , Anticorpos Monoclonais/biossíntese , Anticorpos Monoclonais/efeitos dos fármacos , Anticorpos Monoclonais/metabolismo , Antioxidantes/metabolismo , Reatores Biológicos , Células CHO , Cricetinae , Cricetulus , Cisteína/metabolismo , Cisteína/farmacologia , Glutationa/metabolismo , Glicosilação/efeitos dos fármacos , Concentração de Íons de Hidrogênio , Mapeamento de Peptídeos , Tirosina/metabolismo
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