학술논문

Transient Expression in Red Beet of a Biopharmaceutical Candidate Vaccine for Type-1 Diabetes
Document Type
research-article
Author
Mattia Santoni, author; Edoardo Bertini, author; Roberta Zampieri, author; Anna Cuccurullo, author; Mauro Commisso, author; Elisa Gecchele, author; Linda Avesani, author
Source
Journal of Visualized Experiments. (145)
Subject
Bioengineering
Issue 145
transient expression
edible plant
T1D
oral tolerance
oral vaccine
agroinfiltration
red beet
GAD65
bioequivalence
gastric digestion
Language
English
ISSN
1940-087X
Abstract
Plant molecular farming is the use of plants to produce molecules of interest. In this perspective, plants may be used both as bioreactors for the production and subsequent purification of the final product and for the direct oral delivery of heterologous proteins when using edible plant species. In this work, we present the development of a candidate oral vaccine against Type 1 Diabetes (T1D) in edible plant systems using deconstructed plant virus-based recombinant DNA technology, delivered with vacuum infiltration. Our results show that a red beet is a suitable host for the transient expression of a human derived autoantigen associated to T1D, considered to be a promising candidate as a T1D vaccine. Leaves producing the autoantigen were thoroughly characterized for their resistance to gastric digestion, for the presence of residual bacterial charge and for their secondary metabolic profile, giving an overview of the process production for the potential use of plants for direct oral delivery of a heterologous protein. Our analysis showed almost complete degradation of the freeze-dried candidate oral vaccine following a simulated gastric digestion, suggesting that an encapsulation strategy in the manufacture of the plant-derived GAD vaccine is required.