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J Microbiol Biotechnol ; 29(1): 55-58, 2019 Jan 28.
Artículo en Inglés | MEDLINE | ID: mdl-30394043

RESUMEN

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.


Asunto(s)
Anticuerpos/metabolismo , Técnicas de Cultivo Celular por Lotes/métodos , Dimetilsulfóxido , Electroporación , Animales , Anticuerpos/genética , Células CHO , Técnicas de Cultivo de Célula , Cricetulus , Expresión Génica , Técnicas de Transferencia de Gen , Proteínas Recombinantes de Fusión/genética , Proteínas Recombinantes de Fusión/metabolismo
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