fulltext.study @t Gmail

Characterisation of recombinant CHO cell lines by investigation of protein productivities and genetic parameters

Paper ID Volume ID Publish Year Pages File Format Full-Text
25313 43567 2007 10 PDF Available
Title
Characterisation of recombinant CHO cell lines by investigation of protein productivities and genetic parameters
Abstract

We have generated a recombinant CHO cell line expressing the fusion protein EpoFc. After selection and screening, protein expression, gene and mRNA copy numbers were analysed in order to gain more information on the influence of genetic parameters on the productivity and stability of production cells. Results from semi-quantitative blot methods were compared to quantitative PCR (qPCR) analyses, whose advantage mainly lies in their higher sensitivity, and the cheaper and faster methodology.We developed stable and high producing clones with low gene copy numbers, in contrast to other cell lines where multiple steps of methotrexate amplification have lead to hundreds of copies of inserts with the risk of karyotypic instabilities and decreased growth rates that overcome the benefits of increased productivities.When comparing genetic parameters to productivity, a good correlation of mRNA levels with specific productivity was observed, whereas high gene copy numbers were not always accompanied by high protein expressions. Based on our data derived from a typical example of a cell line development process, genetic parameters are useful tools for the selection of scalable production clones. Nevertheless, a wider range of cell lines has to be investigated in order to implement genetic analyses into a screening process.

Keywords
CHO, Chinese Hamster Ovary; dhfr, dihydrofolate reductase; EpoFc, erythropoietin-huIgG1Fc fusion protein; GCN, gene copy number; MCB, master cell bank; MTX, methotrexate; OPD, orthophenylenediamine; qp, specific productivity; qPCR, quantitative PCR; RT, r
First Page Preview
Characterisation of recombinant CHO cell lines by investigation of protein productivities and genetic parameters
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
Publisher
Database: Elsevier - ScienceDirect
Journal: Journal of Biotechnology - Volume 128, Issue 4, 10 March 2007, Pages 716–725
Authors
, , , , , , ,
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