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

2007 ◽  
Vol 104 (5) ◽  
pp. 403-407 ◽  
Author(s):  
Shinya Yamawaki ◽  
Takehiro Matsumoto ◽  
Yuka Ohnishi ◽  
Yoichi Kumada ◽  
Naofumi Shiomi ◽  
...  
Keyword(s):  
Pichia Pastoris ◽  
Continuous Culture ◽  
Single Chain Variable Fragment ◽  
Single Chain ◽  
Fed Batch ◽  
Feeding Methods ◽  
Methanol Feeding

Effects of methanol feeding methods on chimeric α-amylase expression in continuous culture of Pichia pastoris

2006 ◽  
Vol 101 (3) ◽  
pp. 227-231 ◽  
Author(s):  
Atsushi Nakano ◽  
Chun Yeon Lee ◽  
Arei Yoshida ◽  
Takehiro Matsumoto ◽  
Naofumi Shiomi ◽  
...  
Keyword(s):  
Pichia Pastoris ◽  
Continuous Culture ◽  
Feeding Methods ◽  
Methanol Feeding

Influence of carbon source on cell size and production of anti LDL (-) single-chain variable fragment by a recombinant Pichia pastoris strain

2019 ◽  
Vol 46 (3) ◽  
pp. 3257-3264
Author(s):  
Cesar Andres Diaz Arias ◽  
João Vitor Dutra Molino ◽  
Daniela de Araújo Viana Marques ◽  
Andrea Queiroz Maranhão ◽  
Dulcineia Abdalla Saes Parra ◽  
...  
Keyword(s):  
Pichia Pastoris ◽  
Carbon Source ◽  
Cell Size ◽  
Single Chain Variable Fragment ◽  
Single Chain ◽  
Pastoris Strain ◽  
Recombinant Pichia Pastoris

A new methanol-feeding strategy for the improved production of β-galactosidase in high cell-density fed-batch cultures of Pichia pastoris Mut+ strains

2012 ◽  
Vol 17 (1) ◽  
pp. 76-83 ◽  
Author(s):  
Amir Maghsoudi ◽  
Safoura Hosseini ◽  
Seyed Abbas Shojaosadati ◽  
Ebrahim Vasheghani-Farahani ◽  
Mohsen Nosrati ◽  
...  
Keyword(s):  
Pichia Pastoris ◽  
Cell Density ◽  
High Cell Density ◽  
Feeding Strategy ◽  
High Cell ◽  
Fed Batch ◽  
Batch Cultures ◽  
Methanol Feeding

scholarly journals Controlling methanol feeding for recombinant protein production by Pichia pastoris under oxidation stress in fed-batch fermentation

2020 ◽  
Author(s):  
Rongkang Hu ◽  
Dongming Lan ◽  
Ruiguo Cui ◽  
Haomin Hong ◽  
Yachun Niu ◽  
...  
Keyword(s):  
Oxidative Stress ◽  
Pichia Pastoris ◽  
Batch Fermentation ◽  
Protein Production ◽  
Feeding Rate ◽  
Induction Phase ◽  
Fed Batch ◽  
Ros Accumulation ◽  
Fed Batch Fermentation ◽  
Methanol Feeding

Abstract Background: Methanol can be used by Pichia pastoris as the sole carbon source and inducer to produce recombinant proteins in high-cell-density fermentations, but also damages cells due to reactive oxygen species (ROS) accumulation from methanol oxidation. Here, we study the relationship between methanol feeding and ROS accumulation by controlling constant methanol feeding rate during the induction phase.Results: Higher methanol feeding rate increased the level of ROS accumulation caused by methanol oxidation. While the cell growth rate was proportional to the rate of methanol feeding rate, but maximum total protein production and highest enzyme activity were achieved at methanol feeding rate 4 mL/(L·h) as compared to that with 5 mL/(L·h). Moreover, oxidative demage induced by over accumulation of ROS in P. pastoris during the methanol induction phase caused cell death and reduced protein expression ability. ROS scavenging system analysis reveals that the higher methanol feeding rate, especially 5 mL/(L·h), resulted in increased intracellular catalase activity and decreased glutathione content significantly. Finally, Spearman's correlation analysis further reveals that the reduced glutathione might be beneficial for maintaining cell viability and increasing protein production under oxidative stress caused by toxic accumulation of ROS accumulation.Conclusion: Our findings suggest an integrated strategy to control the feeding of the essential substrate based on analyzing its response to oxidative stress caused by toxic accumulation of ROS accumulation, as well as develop strategy to optimize fed-batch fermentation.

High efficient expression of a functional humanized single-chain variable fragment (scFv) antibody against CD22 in Pichia pastoris

2014 ◽  
Vol 98 (24) ◽  
pp. 10023-10039 ◽  
Author(s):  
Najmeh Zarei ◽  
Behrouz Vaziri ◽  
Mohammad Ali Shokrgozar ◽  
Reza Mahdian ◽  
Ramin Fazel ◽  
...  
Keyword(s):  
Pichia Pastoris ◽  
Scfv Antibody ◽  
Single Chain Variable Fragment ◽  
Single Chain ◽  
High Efficient ◽  
Efficient Expression
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