Venom and Biotherapeutics Molecules Laboratory, Biotechnology Department, Biotechnology Research Center, Pasteur Institute of Iran, Tehran, Iran.
Molecular Biotechnology Laboratory, School of Biotechnology, Institute of Agricultural Technology, Suranaree University of Technology, Nakhon Ratchasima, 30000, Thailand.
Toxicon. 2021 Jul 15;197:106-113. doi: 10.1016/j.toxicon.2021.04.017. Epub 2021 Apr 24.
Snakebite is an important public health problem in tropical and subtropical regions. Macrovipera lebetina is one of the most dangerous snakes in Iran. Envenoming by this snake can lead to respiratory distress, heart attack, bleeding, and death. The specific treatment available is immunized equine serum, which has several side effects like serum sickness. Nowadays, single-chain fragment variable antibodies (scFvs) are one of the fast growing classes of monoclonal antibodies, which are suggested for treatment of envenoming. This study aimed to achieve a fully human scFv antibody against M. lebetina venom from human non-immune library. In this study, scFvs against M. lebetina venom were isolated by phage display technique. Using three rounds of biopanning, two specific scFvs (C37 and C69) with the highest affinity were selected. The selected scFvs purified by nickel affinity chromatography. The specific binding of purified antibodies were confirmed by enzyme-linked immunosorbent assay. The LD as well as HD concentration of the crude venom were obtained to be 45 μg and 120 μg/ml, respectively. C69 neutralized 48% of the hemolysis activity of M. lebetina venom and C37 survived 66% of mice after 115 min of envenoming. Taken together, the results indicate the potential of human non-immune libraries for selection of functional antibodies against M. lebetina venom.
蛇咬伤是热带和亚热带地区的一个重要公共卫生问题。蝮蛇是伊朗最危险的蛇之一。这种蛇的咬伤会导致呼吸困难、心脏病发作、出血和死亡。目前可用的特定治疗方法是免疫马血清，这种血清有几种副作用，如血清病。如今，单链片段可变抗体（scFv）是单克隆抗体中发展最快的一类，被建议用于治疗蛇咬伤。本研究旨在从人源非免疫文库中获得针对 M. lebetina 毒液的完全人源 scFv 抗体。在这项研究中，使用噬菌体展示技术从人源非免疫文库中分离出针对 M. lebetina 毒液的 scFv。经过三轮生物淘选，选择了两种亲和力最高的特异性 scFv（C37 和 C69）。通过镍亲和层析纯化选定的 scFv。酶联免疫吸附试验证实了纯化抗体的特异性结合。粗毒液的 LD 和 HD 浓度分别为 45μg 和 120μg/ml。C69 中和了 M. lebetina 毒液 48%的溶血活性，C37 在中毒后 115 分钟使 66%的小鼠存活。总之，这些结果表明人源非免疫文库在筛选针对 M. lebetina 毒液的功能性抗体方面具有潜力。
