Cultivation of Pichia pastoris carrying the scFv anti LDL (-) antibody fragment. Effect of preculture carbon source
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 genetica...
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doaj-13bc9edf066f40edbb32ee544f7b098e2020-11-24T23:47:16ZengSociedade Brasileira de MicrobiologiaBrazilian Journal of Microbiology1678-440548341942610.1016/j.bjm.2016.11.009S1517-83822017000300419Cultivation of Pichia pastoris carrying the scFv anti LDL (-) antibody fragment. Effect of preculture carbon sourceCesar Andres Diaz AriasDaniela de Araujo Viana MarquesLuciana Pellegrini MalpiediAndrea Queiroz MaranhãoDulcineia Abdalla Saes ParraAttilio ConvertiAdalberto Pessoa JuniorAbstract 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.http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1517-83822017000300419&lng=en&tlng=enPichia pastorisscFv antibody fragmentCryopreservationExpressionCarbon source |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Cesar Andres Diaz Arias Daniela de Araujo Viana Marques Luciana Pellegrini Malpiedi Andrea Queiroz Maranhão Dulcineia Abdalla Saes Parra Attilio Converti Adalberto Pessoa Junior |
spellingShingle |
Cesar Andres Diaz Arias Daniela de Araujo Viana Marques Luciana Pellegrini Malpiedi Andrea Queiroz Maranhão Dulcineia Abdalla Saes Parra Attilio Converti Adalberto Pessoa Junior Cultivation of Pichia pastoris carrying the scFv anti LDL (-) antibody fragment. Effect of preculture carbon source Brazilian Journal of Microbiology Pichia pastoris scFv antibody fragment Cryopreservation Expression Carbon source |
author_facet |
Cesar Andres Diaz Arias Daniela de Araujo Viana Marques Luciana Pellegrini Malpiedi Andrea Queiroz Maranhão Dulcineia Abdalla Saes Parra Attilio Converti Adalberto Pessoa Junior |
author_sort |
Cesar Andres Diaz Arias |
title |
Cultivation of Pichia pastoris carrying the scFv anti LDL (-) antibody fragment. Effect of preculture carbon source |
title_short |
Cultivation of Pichia pastoris carrying the scFv anti LDL (-) antibody fragment. Effect of preculture carbon source |
title_full |
Cultivation of Pichia pastoris carrying the scFv anti LDL (-) antibody fragment. Effect of preculture carbon source |
title_fullStr |
Cultivation of Pichia pastoris carrying the scFv anti LDL (-) antibody fragment. Effect of preculture carbon source |
title_full_unstemmed |
Cultivation of Pichia pastoris carrying the scFv anti LDL (-) antibody fragment. Effect of preculture carbon source |
title_sort |
cultivation of pichia pastoris carrying the scfv anti ldl (-) antibody fragment. effect of preculture carbon source |
publisher |
Sociedade Brasileira de Microbiologia |
series |
Brazilian Journal of Microbiology |
issn |
1678-4405 |
description |
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 8 h. |
topic |
Pichia pastoris scFv antibody fragment Cryopreservation Expression Carbon source |
url |
http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1517-83822017000300419&lng=en&tlng=en |
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