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1.
Biotechnol Lett ; 39(4): 501-509, 2017 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-28054185

RESUMO

OBJECTIVES: To evaluate the characteristics of a novel human cell line, F2N78, including growth performance, physicochemical properties, and biological activity via direct comparison with CHO cells. RESULTS: The culture performance and physicochemical properties of antibodies produced from F2N78 and CHO cells were compared. For charge variants, antibodies produced from F2N78 cells contained a greater acidic charge variants than CHO cells. Regarding main glycoforms, degree of galactosylation was 52% in CT-A produced from F2N78 cells compared to CHO cells (37%). For sialic acid forms, α-2,6-linked sialic acid and N-acetylneuraminic acid (NANA) residues were observed in antibodies produced from F2N78 cells. In contrast, only α-2,3 linked sialic acid forms were detected in antibodies produced from CHO cells, and NANA and N-glycolylneuraminic acid were detected. Hybrid structure and bisecting structure were only observed in F2N78 cells. CONCLUSIONS: F2N78 cells stably produced antibodies with human specific N-glycan. The novel expression system based on human cells may facilitate the development of an alternative host cell for production of recombinant proteins.


Assuntos
Anticorpos Monoclonais/biossíntese , Linhagem Celular/citologia , Glicosilação , Animais , Anticorpos Monoclonais/química , Células CHO , Cromatografia Líquida de Alta Pressão , Cricetulus , Humanos , Células Híbridas/citologia , Ácidos Neuramínicos/química , Polissacarídeos/química , Ácidos Siálicos/química , Espectrometria de Massas por Ionização por Electrospray
2.
J Biosci Bioeng ; 121(3): 317-24, 2016 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-26454770

RESUMO

A human hybrid cell line, F2N78, was developed by somatic fusion of HEK293 and Namalwa cells for the production recombinant biopharmaceutical proteins. In this study, we performed perfusion culture to verify its potential in culture process used for human cell expression platform. Cell viability could be maintained over 90% and high viable cell density was obtained at higher than 1.0 × 10(7) cells/mL by bleeding process in perfusion culture. The cells were adapted well in both culture modes, but there were apparent differences in protein quality. Compared to fed-batch culture, degalactosylated forms such as G0F and G0 as well as Man5 showed no significant increases in perfusion culture. In terms of charge variants, acidic peaks increased, whereas main peaks constantly decreased according to the length of culture period in both methods.


Assuntos
Anticorpos/metabolismo , Técnicas de Cultura Celular por Lotes , Técnicas de Cultura de Células/métodos , Células Híbridas/citologia , Células Híbridas/metabolismo , Proteínas Recombinantes/biossíntese , Contagem de Células , Fusão Celular , Linhagem Celular , Sobrevivência Celular , Células HEK293 , Humanos , Perfusão
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