Shafaghi Mona, Maktoobian Somayeh, Rasouli Rahimeh, Howaizi Nader, Ofoghi Hamideh, Ehsani Parastoo
Department of Molecular Biology, Pasteur Institute of Iran, Tehran, Iran.
Department of Medical Nanotechnology, School of Medicine, International Campus, Tehran University of Medical Sciences, Tehran, Iran.
Iran J Biotechnol. 2018 Apr 18;16(1):e1774. doi: 10.21859/ijb.1774. eCollection 2018 Apr.
Rabies virus is a neurotropic virus that causes fatal, but, a preventable disease in mammals. Administration of rabies immunoglobulin (RIG) is essential for the post-exposure of the prophylaxis to prevent the disease. However, replacement of polyclonal RIGs with alternative monoclonal antibodies (MAbs) that are capable of neutralizing rabies virus has been recommended.
Here, we have investigated the transient expression of the full-size human MAb against rabies virus glycoprotein; the MAb SO57 in the tobacco plants using vacuum agro-infiltration. Previously, stably transformed plants expressing the MAb have been reported.
In this study three vectors carrying the codon-optimized genes for the heavy or light chain and p19 silencing-suppressor were constructed. These vectors were co-infiltrated into leaves and the transgenes were expressed.
Dot blot, Western blotting, ELISA, and in vitro neutralization assays of the plant extracts showed that the human MAb could assemble in tobacco leaves and was able to neutralize rabies virus.
This study is the first report of transient expression of human MAb SO57 gene in tobacco plant within a few days after vacuum agro-infiltration.
狂犬病病毒是一种嗜神经性病毒，可在哺乳动物中引发致命但可预防的疾病。注射狂犬病免疫球蛋白（RIG）对于暴露后预防该疾病至关重要。然而，有人建议用能够中和狂犬病病毒的替代单克隆抗体（MAb）取代多克隆RIG。
在此，我们研究了针对狂犬病病毒糖蛋白的全尺寸人源单克隆抗体；使用真空农杆菌浸润法在烟草植株中瞬时表达单克隆抗体SO57。此前，已有报道称有稳定表达该单克隆抗体的转基因植株。
在本研究中，构建了三个携带重链或轻链密码子优化基因以及p19沉默抑制子的载体。将这些载体共同浸润到叶片中，使转基因得以表达。
对植物提取物进行斑点印迹、蛋白质印迹、酶联免疫吸附测定和体外中和试验表明，人源单克隆抗体能够在烟草叶片中组装并中和狂犬病病毒。
本研究首次报道了在真空农杆菌浸润后几天内，人源单克隆抗体SO57基因在烟草植株中的瞬时表达。
