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J Microbiol Biotechnol ; 29(1): 55-58, 2019 Jan 28.
Artigo em Inglês | MEDLINE | ID: mdl-30394043

RESUMO

Development of stable rCHO cell lines is still time consuming and labor intensive, although it is a critical step in the commercial development of recombinant antibodies. The current work demonstrates, for the first time, that electroporation of CHO cells with DMSO can enhance stable expression of recombinant antibodies in rCHO cells. Electroporation with DMSO resulted in an average 3.7-fold and 2.8-fold increases in expression levels of aflibercept and pembrolizumab, respectively, in pools of stable rCHO cells. It also resulted in an average of 2.2-fold and 2.6-fold increases in the expression of aflibercept and pembrolizumab, respectively, in single-cell derived rCHO clones. Simple batch cultures of rCHO cell clones with the highest expression produced 1.0 g/l for aflibercept and 1.4 g/l for pembrolizumab without a time-consuming gene amplification process. Electroporation with DMSO also shortened the development of rCHO cell lines to 2-3 months, allowing rapid establishment of stable rCHO cell lines with a desirable expression level antibodies.


Assuntos
Anticorpos/metabolismo , Técnicas de Cultura Celular por Lotes/métodos , Dimetil Sulfóxido , Eletroporação , Animais , Anticorpos/genética , Células CHO , Técnicas de Cultura de Células , Cricetulus , Expressão Gênica , Técnicas de Transferência de Genes , Proteínas Recombinantes de Fusão/genética , Proteínas Recombinantes de Fusão/metabolismo
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