Modarresi Mostafa, Javaran Mokhtar Jalali, Shams-Bakhsh Masoud, Zeinali Sirous, Behdani Mahdi, Mirzaee Malihe
1Department of Plant Breeding and Biotechnology, Faculty of Agriculture, Tarbiat Modares University, P.O. Box 14115-336, Tehran, Islamic Republic of Iran.
2Department of Plant Pathology, Faculty of Agriculture, Tarbiat Modares University, Tehran, Islamic Republic of Iran.
3 Biotech. 2018 Dec;8(12):484. doi: 10.1007/s13205-018-1500-z. Epub 2018 Nov 15.
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 (TuMV)-based vector, and then, it was expressed in and 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.
在人类中,血管内皮生长因子(VEGF)与其受体(VEGFR2)之间的相互作用对肿瘤血管生成至关重要。这是癌症肿瘤生长和转移的一个重要过程。通过抗体阻断VEGF/VEGFR2结合可抑制新血管形成和肿瘤转移。本研究旨在利用一个瞬时表达平台在烟草植株中生产重组抗VEGFR2纳米抗体。首先,将抗VEGFR2特异性纳米抗体基因克隆到一个基于烟草花叶病毒(TuMV)的载体中,然后在烟草品种Xanthi中瞬时表达。通过逆转录-聚合酶链反应(RT-PCR)、斑点印迹、酶联免疫吸附测定(ELISA)和蛋白质免疫印迹分析证实了纳米抗体在烟草植株中的表达。结果表明,烟草植株可积累纳米抗体,其含量高达总可溶性蛋白的0.45%(8.3微克/100毫克鲜叶)。这是首次报道利用植物病毒载体在烟草植株中成功表达骆驼源抗VEFGR2纳米抗体基因。该系统为在烟草植株中生产药物和商业蛋白如抗癌纳米抗体提供了一种快速解决方案。