Cultivation of Pichia pastoris carrying the scFv anti LDL (-) antibody fragment. Effect of preculture carbon source

Revista de microbiologia

Malpiedi, Luciana Pellegrini¹  Marques, Daniela de Araujo Viana²  Arias, Cesar Andres Diaz³  Maranhão, Andrea Queiroz⁴ 

[1] Universidade Nacional de Brasíilia, Brasília, Brazil;Universidade Rural Federal de Pernambuco, Recife, Brazil;Universidade de São Paulo, São Paulo, Brazil;University of Genoa, Genoa, Italy

关键词: Pichia pastoris;    scFv antibody fragment;    Cryopreservation;    Expression;    Carbon source.;   

DOI  :  10.1016/j.bjm.2016.11.009

学科分类：农业科学（综合）

来源: Sociedade Brasileira de Microbiologia / Brazilian Society for Microbiology

【 摘 要 】

Antibodies and antibody fragments are nowadays among the most important biotechnological products, and Pichia pastoris is one of the most important vectors to produce them as well as other recombinant proteins. The conditions to effectively cultivate a P. pastoris strain previously genetically modified to produce the single-chain variable fragment anti low density lipoprotein (-) under the control of the alcohol oxidase promoter have been investigated in this study. In particular, it was evaluated if, and eventually how, the carbon source (glucose or glycerol) used in the preculture preceding cryopreservation in 20% glycerol influences both cell and antibody fragment productions either in flasks or in bioreactor. Although in flasks the volumetric productivity of the antibody fragment secreted by cells precultured, cryopreserved and reactivated in glycerol was 42.9% higher compared with cells precultured in glucose, the use of glycerol in bioreactor led to a remarkable shortening of the lag phase, thereby increasing it by no less than thrice compared to flasks. These results are quite promising in comparison with those reported in the literature for possible future industrial applications of this cultivation, taking into account that the overall process time was reduced by around 8 h.

【 授权许可】

CC BY-NC