Cultivation of Pichia Pastoris Carrying the ScFv Anti LDL (-) Antibody Fragment. Effect of Preculture Carbon Source

Overview

Journal Braz J Microbiol

Specialty Microbiology

Date 2017 Feb 27

PMID 28237678

Citations 10

Authors

Cesar Andres Diaz Arias

Daniela de Araujo Viana Marques

Luciana Pellegrini Malpiedi

Andrea Queiroz Maranhao

Dulcineia Abdalla Saes Parra

Attilio Converti

Adalberto Pessoa Junior

Affiliations

Soon will be listed here.

Abstract

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 8h.

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