Transient Expression in Red Beet of a Biopharmaceutical Candidate Vaccine for Type-1 Diabetes
| Autor: | Roberta Zampieri, Anna Cuccurullo, Mattia Santoni, Edoardo Bertini, Elisa Gecchele, Linda Avesani, Mauro Commisso |
|---|---|
| Rok vydání: | 2019 |
| Předmět: |
Agroinfiltration
Swine General Chemical Engineering Green Fluorescent Proteins Administration Oral red beet Heterologous Bioengineering agroinfiltration Biology General Biochemistry Genetics and Molecular Biology transient expression law.invention Spinacia oleracea law Plant virus Metabolome medicine Animals edible plant Issue 145 T1D oral tolerance oral vaccine GAD65 bioequivalence gastric digestion Biological Products Vaccines Type 1 diabetes General Immunology and Microbiology Glutamate Decarboxylase General Neuroscience fungi food and beverages Plants Genetically Modified medicine.disease Genetically modified organism Plant Leaves Diabetes Mellitus Type 1 Biopharmaceutical Biochemistry Mutation Recombinant DNA Beta vulgaris |
| Zdroj: | Journal of Visualized Experiments. |
| ISSN: | 1940-087X |
| DOI: | 10.3791/59298 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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