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

CESAR DÍAZ ARIAS; DANIELA A. VIANA-MARQUEZ; LUCIANA PELLEGRINI MALPIEDI; ANDREA QUEIROZ MARANHÃO; DULCINEIA S.P. ABDALLA; ATTILIO CONVERTI; ADALBERTO PESSOA JR

BRAZILIAN JOURNAL OF MICROBIOLOGY

SOC BRASILEIRA MICROBIOLOGIA

Año: 2017 vol. 48 p. 419 - 426

1517-8382

Antibodies and antibody fragments are nowadays among the most important biotechno-logical products, and Pichia pastoris is one of the most important vectors to produce themas well as other recombinant proteins. The conditions to effectively cultivate a P. pastorisstrain previously genetically modified to produce the single-chain variable fragment antilow density lipoprotein (−) under the control of the alcohol oxidase promoter have beeninvestigated in this study. In particular, it was evaluated if, and eventually how, the carbonsource (glucose or glycerol) used in the preculture preceding cryopreservation in 20% glyc-erol 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 cellsprecultured in glucose, the use of glycerol in bioreactor led to a remarkable shortening ofthe lag phase, thereby increasing it by no less than thrice compared to flasks. These resultsare quite promising in comparison with those reported in the literature for possible futureindustrial applications of this cultivation, taking into account that the overall process timewas reduced by around 8 h.