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Transient expression of anti-VEFGR2 nanobody in Nicotiana tabacum and N. benthamiana

Modarresi, Mostafa, Javaran, Mokhtar Jalali, Shams-bakhsh, Masoud, Zeinali, Sirous, Behdani, Mahdi, Mirzaee, Malihe
3 Biotech 2018 v.8 no.12 pp. 484
Nicotiana benthamiana, Nicotiana tabacum, Turnip mosaic virus, Western blotting, angiogenesis, antibodies, enzyme-linked immunosorbent assay, genes, humans, leaves, metastasis, molecular cloning, neoplasms, proteins, reverse transcriptase polymerase chain reaction, tobacco, vascular endothelial growth factor receptor-2, vascular endothelial growth factors
In human, the interaction between vascular endothelial growth factor (VEGF) and its receptor (VEGFR2) is critical for tumor angiogenesis. This is a vital process for cancer tumor growth and metastasis. Blocking VEGF/VEGFR2 conjugation by antibodies inhibits the neovascularization and tumor metastasis. This investigation designed to use a transient expression platform for production of recombinant anti-VEGFR2 nanobody in tobacco plants. At first, anti-VEGFR2-specific nanobody gene was cloned in a Turnip mosaic virus (TuMV)-based vector, and then, it was expressed in Nicotiana benthamiana and Nicotiana tabacum cv. Xanthi transiently. The expression of nanobody in tobacco plants were confirmed by reverse transcription-polymerase chain reaction (RT-PCR), dot blot, enzyme-linked immunosorbent assays (ELISA), and Western blot analysis. It was shown that tobacco plants could accumulate nanobody up to level 0.45% of total soluble protein (8.3 µg/100 mg of fresh leaf). This is the first report of the successful expression of the camelied anti-VEFGR2 nanobody gene in tobacco plants using a plant viral vector. This system provides a fast solution for production of pharmaceutical and commercial proteins such as anti-cancer nanobodies in tobacco plants.