title: Using matrix attachment regions to improve recombinant protein production.
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.
authors: Laboratory of Molecular Biotechnology, University of Lausanne, Lausanne, Switzerland.
authors: Niamh
authors: N
authors: Harraghy
authors: Montserrat
authors: M
authors: Buceta
authors: Alexandre
authors: A
authors: Regamey
authors: Pierre-Alain
authors: PA
authors: Girod
authors: Nicolas
authors: N
authors: Mermod
mesh_terms: D000818:Animals; D016466:CHO Cells; D003001:Cloning, Molecular; D006224:Cricetinae; D003412:Cricetulus; D004247:DNA / Q000235:genetics / Q000378:metabolism; D004797:Enzyme-Linked Immunosorbent Assay; D005434:Flow Cytometry; D005818:Genetic Engineering / Q000379:methods*; D005822:Genetic Vectors / Q000235:genetics; D049452:Green Fluorescent Proteins / Q000235:genetics; D007074:Immunoglobulin G / Q000096:biosynthesis / Q000235:genetics; D050356:Lipid Metabolism; D045170:Matrix Attachment Regions; D011994:Recombinant Proteins / Q000096:biosynthesis* / Q000235:genetics; D014162:Transfection
chemical_list: D007074:Immunoglobulin G; D011994:Recombinant Proteins; D049452:Green Fluorescent Proteins; D004247:DNA