dc.creatorOcampo, Carolina Gabriela
dc.creatorLareu, Jorge Fabricio
dc.creatorMarín Viegas, Vanesa Soledad
dc.creatorMangano, Silvina
dc.creatorLoos, Andreas
dc.creatorSteinkellner, Herta
dc.creatorPetruccelli, Silvana
dc.date2016-12
dc.date2020-06-11T12:38:20Z
dc.date.accessioned2023-07-14T19:57:05Z
dc.date.available2023-07-14T19:57:05Z
dc.identifierhttp://sedici.unlp.edu.ar/handle/10915/98023
dc.identifierhttps://ri.conicet.gov.ar/11336/57361
dc.identifierhttps://onlinelibrary.wiley.com/doi/full/10.1111/pbi.12580
dc.identifierissn:1467-7644
dc.identifier.urihttps://repositorioslatinoamericanos.uchile.cl/handle/2250/7437323
dc.descriptionPlant-based platforms are extensively used for the expression of recombinant proteins, including monoclonal antibodies. However, to harness the approach effectively and leverage it to its full potential, a better understanding of intracellular processes that affect protein properties is required. In this work, we examined vacuolar (vac) targeting and deposition of the monoclonal antibody (Ab) 14D9 in Nicotiana benthamiana leaves. Two distinct vacuolar targeting signals (KISIA and NIFRGF) were C-terminal fused to the heavy chain of 14D9 (vac-Abs) and compared with secreted and ER-retained variants (sec-Ab, ER-Ab, respectively). Accumulation of ER- and vac-Abs was 10- to 15-fold higher than sec-Ab. N-glycan profiling revealed the predominant presence of plant typical complex fucosylated and xylosylated GnGnXF structures on sec-Ab while vac-Abs carried mainly oligomannosidic (Man 7-9) next to GnGnXF forms. Paucimannosidic glycans (commonly assigned as typical vacuolar) were not detected. Confocal microscopy analysis using RFP fusions showed that sec-Ab-RFP localized in the apoplast while vac-Abs-RFP were exclusively detected in the central vacuole. The data suggest that vac-Abs reached the vacuole by two different pathways: direct transport from the ER bypassing the Golgi (Ab molecules containing Man structures) and trafficking through the Golgi (for Ab molecules containing complex N-glycans). Importantly, vac-Abs were correctly assembled and functionally active. Collectively, we show that the central vacuole is an appropriate compartment for the efficient production of Abs with appropriate post-translational modifications, but also point to a reconsideration of current concepts in plant glycan processing.
dc.descriptionCentro de Investigación y Desarrollo en Criotecnología de Alimentos
dc.formatapplication/pdf
dc.format2265-2275
dc.languageen
dc.rightshttp://creativecommons.org/licenses/by/4.0/
dc.rightsCreative Commons Attribution 4.0 International (CC BY 4.0)
dc.subjectBiología
dc.subjectQuímica
dc.subjectImmunoglobulin
dc.subjectMolecular farming
dc.subjectN-glycosylation
dc.subjectSecretory pathway
dc.subjectVacuolar sorting signals
dc.subjectVacuolar transport
dc.titleVacuolar targeting of recombinant antibodies in Nicotiana benthamiana
dc.typeArticulo
dc.typeArticulo


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