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A GFP-based method facilitates clonal selection of transfected CHO cells

Title data

Freimark, Denise ; Jérôme, Valérie ; Freitag, Ruth:
A GFP-based method facilitates clonal selection of transfected CHO cells.
In: Biotechnology Journal. Vol. 5 (January 2010) Issue 1 . - pp. 24-31.
ISSN 1860-7314
DOI: https://doi.org/10.1002/biot.200800264

Official URL: Volltext

Abstract in another language

The identification of highly expressing clones is a crucial step in the development of cell lines for production of recombinant proteins. Here we present a method based on the co-expression of enhanced green fluorescent protein (EGFP) that allows clonal selection in standard 96-well cell culture plates. The genes encoding the EGFP protein and the protein of interest are linked by an internal ribosome entry site and thus are transcribed into the same mRNA but are translated independently. Since both proteins arise from a common mRNA, the EGFP expression level correlates with the expression level of the therapeutic protein for each clone. By expressing recombinant growth factors in CHO cells, we demonstrate the robustness and performance of this technique. The method is an alternative to the identification of high-producer clones using various cell sorting methods, as it can be performed with standard laboratory equipment.

Further data

Item Type: Article in a journal
Refereed: Yes
Keywords: Growth factor; High producer; Mammalian cell culture; Recombinant protein production; Transfection
Institutions of the University: Faculties > Faculty of Engineering Science
Faculties > Faculty of Engineering Science > Chair Process Biotechnology
Faculties > Faculty of Engineering Science > Chair Process Biotechnology > Chair Process Biotechnology - Univ.-Prof. Dr. Ruth Freitag
Faculties
Result of work at the UBT: Yes
DDC Subjects: 500 Science
500 Science > 500 Natural sciences
600 Technology, medicine, applied sciences
600 Technology, medicine, applied sciences > 620 Engineering
Date Deposited: 24 Feb 2016 13:17
Last Modified: 24 Feb 2016 13:17
URI: https://eref.uni-bayreuth.de/id/eprint/31030