Large-scale transfection of mammalian cells for the fast production of recombinant protein.
Recombinant proteins (r-proteins) are increasingly important in fundamental research and for clinical applications. As many of these r-proteins are of human or animal origin, cultivated mammalian cells are the host of choice to ensure their functional folding and proper posttranslational modifications. Large-scale transfection of human embryonic kidney 293 or Chinese hamster ovary cells is now an established technology that can be used in the production of hundreds of milligram to gram quantities of a r-protein in less than 1 mo from cloning of its cDNA. This chapter aims to provide an overview of large-scale transfection technology with a particular emphasis on calcium phosphate and polyethylenimine-mediated gene transfer.
Pham
Phuong Lan
PL
Laboratoire de Biotechnologie Vétérinaire et Alimentaire, Faculté de Médecine Vétérinaire, Université de Montréal, CP5000, Sainte-Hyacinthe (Québec) J2S 7C6, Canada.
Kamen
Amine
A
Durocher
Yves
Y
D000818:Animals; D001709:Biotechnology / Q000379:methods*; D002130:Calcium Phosphates / Q000494:pharmacology; D002460:Cell Line; D006224:Cricetinae; D003470:Culture Media; D005822:Genetic Vectors / Q000235:genetics*; D006801:Humans; D011094:Polyethyleneimine / Q000494:pharmacology; D011994:Recombinant Proteins / Q000096:biosynthesis*; D014162:Transfection / Q000379:methods*
D002130:Calcium Phosphates; D003470:Culture Media; D011994:Recombinant Proteins; D011094:Polyethyleneimine; C020243:calcium phosphate