Department of Biotechnology and Biomedicine, Technical University of Denmark, DK-2800 Kongens Lyngby, Denmark.
Sophion Bioscience A/S, DK-2750 Ballerup, Denmark.
N Biotechnol. 2023 Sep 25;76:23-32. doi: 10.1016/j.nbt.2023.04.002. Epub 2023 Apr 8.
Phage display technology is a powerful tool for selecting monoclonal antibodies against a diverse set of antigens. Within toxinology, however, it remains challenging to generate monoclonal antibodies against many animal toxins, as they are difficult to obtain from venom. Recombinant toxins have been proposed as a solution to overcome this challenge, but so far, few have been used as antigens to generate neutralizing antibodies. Here, we describe the recombinant expression of α-cobratoxin in E. coli and its successful application as an antigen in a phage display selection campaign. From this campaign, an scFv (single-chain variable fragment) was isolated with similar binding affinity to a control scFv generated against the native toxin. The selected scFv recognizes a structural epitope, enabling it to inhibit the interaction between the acetylcholine receptor and the native toxin in vitro. This approach represents the first entirely in vitro antibody selection strategy for generating neutralizing monoclonal antibodies against a snake toxin.
噬菌体展示技术是一种从多样化抗原中筛选单克隆抗体的强大工具。然而，在毒素学领域，由于从毒液中获得许多动物毒素较为困难，因此针对这些毒素生成单克隆抗体仍然具有挑战性。人们提出使用重组毒素作为克服这一挑战的解决方案，但迄今为止，很少有重组毒素被用作抗原来产生中和抗体。在这里，我们描述了α-眼镜蛇毒素在大肠杆菌中的重组表达，并成功将其作为抗原应用于噬菌体展示筛选实验中。从该筛选实验中，分离到了一个 scFv（单链可变片段），其与针对天然毒素生成的对照 scFv 具有相似的结合亲和力。该 scFv 识别一个结构表位，使其能够在体外抑制乙酰胆碱受体与天然毒素之间的相互作用。这种方法代表了针对蛇毒素生成中和性单克隆抗体的首例完全体外抗体筛选策略。
