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Optimised mammalian expression through the coupling of codon adaptation with gene amplification: Maximum yields with minimum effort

Paper ID Volume ID Publish Year Pages File Format Full-Text
24388 43511 2010 8 PDF Available
Title
Optimised mammalian expression through the coupling of codon adaptation with gene amplification: Maximum yields with minimum effort
Abstract

Gene amplification methodologies are frequently employed for the generation of large quantities of recombinant proteins in mammalian cells. Although they usually guarantee very high yields, they are very time consuming. In addition, due to the large genomic re-arrangements that frequently occur with amplification, the resulting high-producing clones can be unstable. We herein describe significant improvements to the dihydrofolate reductase (DHFR)/methotrexate (MTX) based gene amplification methodology typically employed to improve yields of recombinant proteins produced in genetically engineered CHO host cells. We demonstrate substantial synergy when such gene amplification is combined with extremely high codon optimisation strategies. As a result, expression saturation can be achieved rapidly, in as low as 5 nM MTX, with minimal effort and without compromise in final yields achieved.

Keywords
Gene amplification; CHO; Methotrexate; Codon adaptation; CAI; Antibody expression
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Optimised mammalian expression through the coupling of codon adaptation with gene amplification: Maximum yields with minimum effort
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Publisher
Database: Elsevier - ScienceDirect
Journal: Journal of Biotechnology - Volume 146, Issue 4, 15 April 2010, Pages 186–193
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