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Application of destabilizing sequences on selection marker for improved recombinant protein productivity in CHO-DG44

Paper ID Volume ID Publish Year Pages File Format Full-Text
31991 44867 2007 13 PDF Available
Title
Application of destabilizing sequences on selection marker for improved recombinant protein productivity in CHO-DG44
Abstract

We have developed a systematic and generic way to improve recombinant protein productivities in stable transfections by applying mRNA and protein destabilizing elements to reduce selection marker expression strength. Interferon-gamma (IFNγ) expression vectors containing different combinations of AU-rich elements (ARE) and mouse ornithine decarboxylase (MODC) PEST region on the amplifiable dihydrofolate reductase (dhfr) selection marker were stably transfected into CHO-DG44 cells. Improvements in specific IFNγ productivities were 1.7-, 6.6- and 13.3-fold with the application of ARE, MODC PEST, and both ARE and MODC PEST, respectively. To further enhance productivities, compatibility of the destabilizing sequences with methotrexate (MTX) amplification was validated by amplifying the transfected cells to 50 nM MTX. A 14- to 27-fold increase in specific IFNγ productivities were observed after amplification, indicating the compatibility of the two systems. A high specific IFNγ productivity of 1.05 pg/cell/day was also attained by the amplified cell pool with both ARE and MODC PEST.

Keywords
CHO-DG44; Mammalian cell culture; Recombinant protein expression; Interferon-gamma; Dihydrofolate reductase; Methotrexate amplification; Murine ornithine decarboxylase; PEST region; AU-rich elements
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Application of destabilizing sequences on selection marker for improved recombinant protein productivity in CHO-DG44
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Publisher
Database: Elsevier - ScienceDirect
Journal: Metabolic Engineering - Volume 9, Issue 3, May 2007, Pages 304–316
Authors
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Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering
Get Full-Text Now
Don't Miss Today's Special Offer
Price was $35.95
You save - $31
Price after discount Only $4.95
100% Money Back Guarantee
Full-text PDF Download
Online Support
Any Questions? feel free to contact us