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BMC Biotechnol ; 17(1): 18, 2017 02 22.
Artigo em Inglês | MEDLINE | ID: mdl-28228095

RESUMO

BACKGROUND: As the demand for monoclonal antibodies (mAb) increases, more efficient expression methods are required for their manufacturing process. Transcriptional gene silencing is a common phenomenon in recombinant cell lines which leads to expression reduction and instability. There are reports on improved antibody expression in ubiquitous chromatin opening element (UCOE) containing both heavy and light chain gene constructs. Here we investigate the impact of having these elements as part of the light chain, heavy chain or both genes during cell line development. In this regard, non-UCOE and UCOE vectors were constructed and stable Chinese hamster ovary (CHO) cell pools were generated by different vector combinations. RESULTS: Expression analysis revealed that all UCOE cell pools had higher antibody yields compared to non-UCOE cells, Moreover the most optimal expression was obtained by cells containing just the UCOE on heavy chain. In terms of stability, it was shown that the high level of expression was kept consistence for more than four months in these cells whereas the expression titers were reduced in the other UCOE pools. CONCLUSIONS: In conclusion, UCOE significantly enhanced the level and stability of antibody expression and the use of this element with heavy chain provided more stable cell lines with higher production level.


Assuntos
Anticorpos Monoclonais/biossíntese , Anticorpos Monoclonais/genética , Cromatina/genética , Regulação da Expressão Gênica/genética , Vetores Genéticos/genética , Engenharia de Proteínas/métodos , Animais , Células CHO , Cricetulus , Regiões Promotoras Genéticas/genética , Transgenes/genética
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