Efficient production of recombinant IgG by metabolic control and co-expression with GLUT5 in a fructose-based medium.
A fructose-based cell culture is suitable for the process control of protein production because of slow sugar consumption rate and low lactate accumulation. The fructose transporter, GLUT5, mediates its incorporation into cells and is required for the fructose-based culture. In order to produce efficiently recombinant IgG by metabolic control and co-expression with GLUT5 in a fructose-based medium, an IgG and GLUT5 co-expression vector was constructed and transfected into the human myeloma derived cell line, SC-01MFP, which produced stably recombinant proteins. The cell proliferation in the fructose-based medium was improved by the GLUT5 gene transfection. The recombinant IgG production of the cells cultured in the fructose-based medium exhibited about two-fold increase of that in the glucose-based medium. Flow cytometoric analysis indicated that the GLUT5 protein expression level in cell surface was increased in the fructose-based medium. An exogenous but not endogenous GLUT5 transcription activator remarkably raised IgG productivity in the fructose-based medium when compared to that in the glucose-based medium, suggesting that exogenous GLUT5 expression may be involved in it. The GLUT5 co-expression system may be useful for efficient production of recombinant proteins by the fructose-based cell culture.
Inoue
Yuichi
Y
The Cell Engineering Center, Kitakyushu National College of Technology, 5-20-1 Shii, Kokuraminami-ku, Kitakyushu-shi, Fukuoka, 802-0985, Japan, inoue@kct.ac.jp.
Tsukamoto
Yuriko
Y
Yamanaka
Makoto
M
Nakamura
Shigeki
S
Inoue
Aiko
A
Nishino
Norikazu
N
Kawahara
Hiroharu
H