The Donnelly Centre, University of Toronto, Toronto, Ontario, Canada.
Department of Biotechnology and Biomedicine, Technical University of Denmark, Kongens Lyngby, Denmark.
Protein Sci. 2022 May;31(5):e4296. doi: 10.1002/pro.4296.
Each year, thousands of people fall victim to envenomings caused by cobras. These incidents often result in death due to paralysis caused by α-neurotoxins from the three-finger toxin (3FTx) family, which are abundant in elapid venoms. Due to their small size, 3FTxs are among the snake toxins that are most poorly neutralized by current antivenoms, which are based on polyclonal antibodies of equine or ovine origin. While antivenoms have saved countless lives since their development in the late 18th century, an opportunity now exists to improve snakebite envenoming therapy via the application of new biotechnological methods, particularly by developing monoclonal antibodies against poorly neutralized α-neurotoxins. Here, we describe the use of phage-displayed synthetic antibody libraries and the development and characterization of six synthetic antibodies built on a human IgG framework and developed against α-cobratoxin - the most abundant long-chain α-neurotoxin from Naja kaouthia venom. The synthetic antibodies exhibited sub-nanomolar affinities to α-cobratoxin and neutralized the curare-mimetic effect of the toxin in vitro. These results demonstrate that phage display technology based on synthetic repertoires can be used to rapidly develop human antibodies with drug-grade potencies as inhibitors of venom toxins.
每年,都有成千上万的人因被眼镜蛇咬伤而中毒。这些事件常常导致死亡,原因是三指毒素(3FTx)家族的α-神经毒素引起的瘫痪,而这种毒素在各种蛇毒中含量丰富。由于 3FTx 体积小,因此是目前基于马或羊源多克隆抗体的抗蛇毒血清中和效果最差的蛇毒之一。自 18 世纪末抗蛇毒血清问世以来,它已挽救了无数生命,但现在有机会通过应用新的生物技术方法来改善蛇咬伤中毒的治疗效果,特别是通过开发针对中和效果不佳的α-神经毒素的单克隆抗体。在这里,我们描述了使用噬菌体展示的合成抗体文库以及基于人 IgG 框架开发和表征的六种针对α-眼镜蛇毒素(来自 Naja kaouthia 毒液的最丰富的长链α-神经毒素)的合成抗体。这些合成抗体对α-眼镜蛇毒素表现出亚纳摩尔亲和力,并在体外中和了毒素的箭毒样作用。这些结果表明,基于合成库的噬菌体展示技术可用于快速开发具有药物效力的人源抗体,作为毒液毒素的抑制剂。