Protocol

Protein Expression in Mammalian Cells

Volume 801 of the series Methods in Molecular Biology pp 93-110

Date:

Using Matrix Attachment Regions to Improve Recombinant Protein Production

  • Niamh HarraghyAffiliated withLaboratory of Molecular Biotechnology, University of Lausanne
  • , Montserrat BucetaAffiliated withSelexis SA
  • , Alexandre RegameyAffiliated withSelexis SA
  • , Pierre-Alain GirodAffiliated withSelexis SA
  • , Nicolas MermodAffiliated withLaboratory of Molecular Biotechnology, University of Lausanne Email author 

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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.

Key words

Matrix attachment region Recombinant protein production Chinese hamster ovary cells Antibody IgG ELISA