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

• Published in: Journal of bioscience and bioengineering 2007-11, Vol.104 (5), p.403-407
• Main Authors: Yamawaki, Shinya, Matsumoto, Takehiro, Ohnishi, Yuka, Kumada, Yoichi, Shiomi, Naofumi, Katsuda, Tomohisa, Lee, Eun Kyu, Katoh, Shigeo
• Format: Article
• Language: English
• Subjects: ALIMENTACION; ALIMENTATION; ANTIBODIES; ANTICORPS; ANTICUERPOS; Biological and medical sciences; Biotechnology; Cell Culture Techniques - methods; continuous culture; DO-stat; fed-batch culture; FEEDING; Fermentation; Fundamental and applied biological sciences. Psychology; Immunoglobulin Fab Fragments - biosynthesis; Immunoglobulin Fab Fragments - genetics; Immunoglobulin Variable Region - biosynthesis; Immunoglobulin Variable Region - genetics; Industrial Microbiology; METANOL; METHANOL; Methanol - metabolism; Methanol - pharmacology; methanol concentration control; methylotrophic yeast; Pichia - drug effects; Pichia - growth & development; PICHIA PASTORIS; Recombinant Proteins - biosynthesis; Recombinant Proteins - genetics; scFv
• ISSN: 1389-1723; 1347-4421
• DOI: 10.1263/jbb.104.403

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.