Production of Single-Chain Variable Fragment Antibody (scFv) in Fed-Batch and Continuous Culture of Pichia pastoris by Two Different Methanol Feeding Methods(BIOCHEMICAL ENGINEERING)

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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 3.9g l^<-1> in fed-batch culture, a scFv concentration of 198mg 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 810mg l^<-1> at a dilution rate of 0.0094 h^<-1> was obtained by maintaining the methanol concentration at 3.9g 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.
社団法人日本生物工学会の論文
2007-11-25