Overview of Protein Expression by Mammalian Cells

Abstract

This unit reviews the stages involved in protein production in mammalian cells using a stable-expression approach. Choice of cell type is discussed, as is transfection of the host cells, methods for selection and amplification of transformants, and growth of cells at appropriate scale for protein production. Since post-transcriptional modification and intracellular protein transportation are important features of recombinant-protein production in mammalian cells, some description of these mechanisms is included.

Novel human cell expression method reveals the role and prevalence of posttranslational modification in nonmuscle tropomyosins

“…After purification on a high-affinity chitin resin, self-cleavage of the intein takes place precisely after the last endogenous amino acid of the expressed target protein, leaving no extra amino acids from the tag. While both mammalian cell expression ( 59 , 60 , 61 ) and intein-CBD purification tags ( 62 ) have been previously used, to our knowledge this is the first time that the two are combined in a single method. Self-cleavage of the intein is thiol-catalyzed, meaning that a portion of the intein of bacterial origin is prematurely activated in the reducing environment of the cytoplasm of eukaryotic cells, likely explaining why they have not been used for mammalian cell expression.…”

Recombinant Feline Oviductin – A Powerful Tool for Functional IVF Studies in the Domestic Cat

“…N ‐glycosylation and O ‐glycosylation occur also in the ER or the early part of the Golgi complex. Misfolded proteins are immediately retained and eventually degraded (Gray ). Only correctly folded and glycosylated proteins are secreted to the medium.…”

“…It is known that a number of proteins are not normally secreted by mammalian cells. Recombinant protein is retained in the cell or on the cell surface or accumulates in the Golgi or ER (Gray ).…”

Humoral Immune Response in a Mouse Model Induced With Dengue Virus-like Particles Serotypes 1 and 4 Produced in Silkworm Larvae

“…While mammalian cells secrete proteins with correct conformations and full biological activity, insect cells offer advantages comparable to those of mammalian cells when compared to E. coli or yeast expression systems. The ability to introduce foreign DNA into these cells facilitates a better understanding of the transcriptional, translational, and posttranslational machinery in mammalian cells (Gray 2001;Ikonomou et al 2003).…”