title: Fundamentals of Expression in Mammalian Cells.
abstract: Expression of proteins in mammalian cells is a key technology important for many functional studies on human and higher eukaryotic genes. Studies include the mapping of protein interactions, solving protein structure by crystallization and X-ray diffraction or solution phase NMR and the generation of antibodies to enable a range of studies to be performed including protein detection in vivo. In addition the production of therapeutic proteins and antibodies, now a multi billion dollar industry, has driven major advances in cell line engineering for the production of grams per liter of active proteins and antibodies. Here the key factors that need to be considered for successful expression in HEK293 and CHO cells are reviewed including host cells, expression vector design, transient transfection methods, stable cell line generation and cultivation conditions.
authors.affiliation: IONTAS Ltd, Babraham Research Campus, Babraham, Cambridge, CB22 3AT, UK. mrd@iontas.co.uk.
authors.forename: Michael R
authors.initials: MR
authors.lastname: Dyson
mesh_terms: D000818:Animals; D016466:CHO Cells; D018929:Cell Culture Techniques; D006224:Cricetinae; D003412:Cricetulus; D005786:Gene Expression Regulation; D005822:Genetic Vectors; D057809:HEK293 Cells; D057166:High-Throughput Screening Assays; D006801:Humans; D046912:Multiprotein Complexes; D015202:Protein Engineering / Q000379:methods*; D055503:Protein Multimerization; D020836:Protein Structure, Quaternary; D021122:Protein Subunits; D011994:Recombinant Proteins / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics; D013329:Structure-Activity Relationship; D014158:Transcription, Genetic; D014162:Transfection
chemical_list: D046912:Multiprotein Complexes; D021122:Protein Subunits; D011994:Recombinant Proteins
keywords: Antibody expression; Biologics production; High-throughput expression; Transient transfection