Expression and purification of the epidermal growth factor receptor extracellular domain utilizing a polycistronic expression system.
Protein Expr Purif
; 4(3): 177-86, 1993 Jun.
Article
em En
| MEDLINE
| ID: mdl-8518559
ABSTRACT
High level expression of the epidermal growth factor receptor ectodomain (EGFR-ED) has been achieved using a polycistronic expression system. The expression vector was designed such that EGFR-ED was at the 5' end of a tricistron followed by luciferase and dihydrofolate reductase (dhfr). Following transfection into Chinese hamster ovary cells, clones were isolated under selective conditions for dhfr expression and monitored for luciferase activity and EGFR-ED expression using immunofluorescence microscopy. A 100-kDa protein corresponding to EGFR-ED was efficiently secreted from expressing cells. Two purification schemes were used to obtain protein at least 95% pure. Glutaraldehyde crosslinking was used to show that EGFR-ED specifically binds EGF and TGF alpha and that the affinity for EGF is 5.5 x 10(-7) M.
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Base de dados:
MEDLINE
Assunto principal:
Receptores ErbB
/
Vetores Genéticos
Limite:
Animals
/
Humans
Idioma:
En
Revista:
Protein Expr Purif
Assunto da revista:
BIOLOGIA MOLECULAR
Ano de publicação:
1993
Tipo de documento:
Article