Production of single-chain variable fragment antibody (scFv) in fed-batch and continuous culture of Pichia pastoris by two different methanol feeding methods.
This study examined the effects of two methods of methanol feeding, DO-stat and methanol concentration control, in fed-batch and continuous cultures of Pichia pastoris on cell growth and single-chain variable fragment antibody (scFv) expression. By maintaining the methanol concentration at 3.9 g l(-1) in fed-batch culture, a scFv concentration of 198 mg l(-1) was obtained. In continuous culture using both methanol feeding methods, the scFv concentration in the fermentation broth increased with a decreasing dilution rate. A maximum scFv concentration of 810 mg l(-1) at a dilution rate of 0.0094 h(-1) was obtained by maintaining the methanol concentration at 3.9 g l(-1). Although the specific methanol consumption rate was the same for both methods, the specific productivity of scFv was higher in methanol concentration control from 0.0094 to 0.049 h(-1) than it was in DO-stat control. Therefore, continuous culture with methanol feeding by the concentration control method shows promise for the industrial scale production of recombinant proteins by Pichia pastoris.
Yamawaki
Shinya
S
Department of Chemical Science and Engineering, Kobe University, 1-1 Rokkodai, Nada, Kobe 657-8501, Japan.
Matsumoto
Takehiro
T
Ohnishi
Yuka
Y
Kumada
Yoichi
Y
Shiomi
Naofumi
N
Katsuda
Tomohisa
T
Lee
Eun Kyu
EK
Katoh
Shigeo
S
D018929:Cell Culture Techniques / Q000379:methods*; D005285:Fermentation; D007140:Immunoglobulin Fab Fragments / Q000096:biosynthesis* / Q000235:genetics; D007135:Immunoglobulin Variable Region / Q000096:biosynthesis* / Q000235:genetics; D007218:Industrial Microbiology; D000432:Methanol / Q000378:metabolism* / Q000494:pharmacology; D010843:Pichia / Q000187:drug effects / Q000254:growth & development*; D011994:Recombinant Proteins / Q000096:biosynthesis* / Q000235:genetics
D007140:Immunoglobulin Fab Fragments; D007135:Immunoglobulin Variable Region; D011994:Recombinant Proteins; D000432:Methanol