Using matrix attachment regions to improve recombinant protein production.

Details

Serval ID
serval:BIB_96D3473CAF1E
Type
Article: article from journal or magazin.
Collection
Publications
Title
Using matrix attachment regions to improve recombinant protein production.
Journal
Methods in Molecular Biology
Author(s)
Harraghy N., Buceta M., Regamey A., Girod P.A., Mermod N.
ISSN
1940-6029 (Electronic)
ISSN-L
1064-3745
Publication state
Published
Issued date
2012
Volume
801
Pages
93-110
Language
english
Abstract
Chinese hamster ovary (CHO) cells are the system of choice for the production of complex molecules, such as monoclonal antibodies. Despite significant progress in improving the yield from these cells, the process to the selection, identification, and maintenance of high-producing cell lines remains cumbersome, time consuming, and often of uncertain outcome. Matrix attachment regions (MARs) are DNA sequences that help generate and maintain an open chromatin domain that is favourable to transcription and may also facilitate the integration of several copies of the transgene. By incorporating MARs into expression vectors, an increase in the proportion of high-producer cells as well as an increase in protein production are seen, thereby reducing the number of clones to be screened and time to production by as much as 9 months. In this chapter, we describe how MARs can be used to increase transgene expression and provide protocols for the transfection of CHO cells in suspension and detection of high-producing antibody cell clones.
Keywords
Animals, CHO Cells, Cloning, Molecular, Cricetinae, Cricetulus, DNA/genetics, DNA/metabolism, Enzyme-Linked Immunosorbent Assay, Flow Cytometry, Genetic Engineering/methods, Genetic Vectors/genetics, Green Fluorescent Proteins/genetics, Immunoglobulin G/biosynthesis, Immunoglobulin G/genetics, Lipid Metabolism, Matrix Attachment Regions, Recombinant Proteins/biosynthesis, Recombinant Proteins/genetics, Transfection
Pubmed
Web of science
Create date
24/02/2011 12:24
Last modification date
20/08/2019 15:58
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