Recombinant glycoprotein product quality in proliferation-controlled BHK-21 cells.
We analyzed product quality to determine the applicability of proliferation-controlled mammalian cells for recombinant pharmaceutical protein production. Baby hamster kidney (BHK)-21 cells were engineered to express a dicistronic, stabilized, self-selecting growth control system consisting of a beta-estradiol-activatable transcription factor IRF-1 fusion protein. IRF-1 activity led to a reduced growth rate, whereas productivity, protein integrity, and glycosylation pattern of the industrially relevant secreted pharmaceutical glycoprotein erythropoietin remained consistent, showing that this technique has the potential for improving the consistency of high-quality pharmaceutical products and thus warrants further development.
Mueller
P P
PP
Department of Gene Regulation and Differentiation, GBF-National Research Center for Biotechnology, Mascheroder Weg 1, D-38124 Braunschweig, Germany. PMU@gbf.de
Schlenke
P
P
Nimtz
M
M
Conradt
H S
HS
Hauser
H
H
D000818:Animals; D001709:Biotechnology; D002240:Carbohydrate Sequence; D002455:Cell Division; D002460:Cell Line; D006224:Cricetinae; D004268:DNA-Binding Proteins / Q000235:genetics; D004921:Erythropoietin / Q000235:genetics / Q000302:isolation & purification / Q000592:standards; D006023:Glycoproteins / Q000096:biosynthesis / Q000592:standards; D006031:Glycosylation; D006801:Humans; D050856:Interferon Regulatory Factor-1; D008969:Molecular Sequence Data; D009844:Oligosaccharides / Q000737:chemistry / Q000302:isolation & purification; D010750:Phosphoproteins / Q000235:genetics; D015202:Protein Engineering; D011786:Quality Control; D011994:Recombinant Proteins / Q000096:biosynthesis* / Q000235:genetics / Q000592:standards
D004268:DNA-Binding Proteins; D006023:Glycoproteins; C494257:IRF1 protein, human; D050856:Interferon Regulatory Factor-1; D009844:Oligosaccharides; D010750:Phosphoproteins; D011994:Recombinant Proteins; D004921:Erythropoietin