High level expression of proteins using sequences from the ferritin heavy chain gene locus.
An expression vector has been generated using a gene highly expressed under conditions found in a typical fed-batch bioreactor process. The ferritin heavy chain (HC) gene exhibits higher levels of expression in the late stages of a fed-batch bioreactor than in the early stages. This property was considered advantageous for an expression vector, since the maximal cell density would coincide with maximal expression. The rat ferritin HC genomic region was isolated and converted into an expression vector where large segments of 5' and 3' flanking regions were included in an attempt to recreate the same high level of expression in stably transfected cells. Expression from the resulting ferritin HC vector was compared to vectors containing the commonly used strong promoters, CMV IE, and SV40 early promoter/enhancer, in the generation of stable transfectants. The ferritin HC vector was able to generate cell lines with significantly higher expression levels than those under the control of the viral promoters.
Prentice
Holly L
HL
Momenta Pharmaceuticals, Inc., Cambridge, MA 02142, United States. hprentice@momentapharma.com
Tonkin
Christopher J D
CJ
Caamano
Louisa
L
Sisk
William P
WP
D000818:Animals; D001052:Apoferritins / Q000235:genetics* / Q000378:metabolism; D001483:Base Sequence; D019149:Bioreactors; D016466:CHO Cells; D006224:Cricetinae; D003412:Cricetulus; D005786:Gene Expression Regulation / Q000235:genetics*; D005822:Genetic Vectors / Q000096:biosynthesis / Q000235:genetics*; D008969:Molecular Sequence Data; D051381:Rats; D016196:S Phase / Q000235:genetics / Q000502:physiology
D001052:Apoferritins