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

Autor:	Arias CAD; Universidade de São Paulo, Departamento Bioquímico - Tecnologia Farmacêutica, São Paulo, SP, Brazil., Marques DAV; Universidade Rural Federal de Pernambuco, Departamento de Morfologia e Fisiologia Animal, Recife, PE, Brazil., Malpiedi LP; Universidade de São Paulo, Departamento Bioquímico - Tecnologia Farmacêutica, São Paulo, SP, Brazil., Maranhão AQ; Universidade Nacional de Brasíilia, Departamento de Biologia Celular, Laboratório de Imunologia Molecular, Brasília, DF, Brazil., Parra DAS; Universidade de São Paulo, Departamento de Análises Clínicas e Toxicologia, São Paulo, SP, Brazil., Converti A; University of Genoa, Department of Civil, Chemical and Environmental Engineering, Genoa, Italy. Electronic address: converti@unige.it., Junior AP; Universidade de São Paulo, Departamento Bioquímico - Tecnologia Farmacêutica, São Paulo, SP, Brazil.
Jazyk:	angličtina
Zdroj:	Brazilian journal of microbiology : [publication of the Brazilian Society for Microbiology] [Braz J Microbiol] 2017 Jul - Sep; Vol. 48 (3), pp. 419-426. Date of Electronic Publication: 2017 Feb 09.
DOI:	10.1016/j.bjm.2016.11.009
Abstrakt:	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. (Copyright © 2017 Sociedade Brasileira de Microbiologia. Published by Elsevier Editora Ltda. All rights reserved.)
Databáze:	MEDLINE
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