Department of Biotechnology and Biomedicine, Technical University of Denmark, 2800, Kongens Lyngby, Denmark.
Sci Rep. 2023 Jun 22;13(1):10181. doi: 10.1038/s41598-023-37056-6.
Antibodies with cross-reactive binding and broad toxin-neutralizing capabilities are advantageous for treating indications such as infectious diseases and animal envenomings. Such antibodies have been successfully selected against closely related antigens using phage display technology. However, the mechanisms driving antibody cross-reactivity typically remain to be elucidated. Therefore, we sought to explore how a previously reported phage display-based cross-panning strategy drives the selection of cross-reactive antibodies using seven different snake toxins belonging to three protein (sub-)families: phospholipases A, long-chain α-neurotoxins, and short-chain α-neurotoxins. We showcase how cross-panning can increase the chances of discovering cross-reactive single-chain variable fragments (scFvs) from phage display campaigns. Further, we find that the feasibility of discovering cross-reactive antibodies using cross-panning cannot easily be predicted by analyzing the sequence, structural, or surface similarity of the antigens alone. However, when antigens share the (exact) same functions, this seems to increase the chances of selecting cross-reactive antibodies, which may possibly be due to the existence of structurally similar motifs on the antigens.
具有交叉反应性结合和广泛中和毒素能力的抗体对于治疗传染病和动物中毒等适应症非常有利。已经使用噬菌体展示技术成功地针对密切相关的抗原选择了这种抗体。然而，驱动抗体交叉反应性的机制通常仍需阐明。因此，我们试图探索以前报道的基于噬菌体展示的交叉筛选策略如何利用属于三种蛋白质（亚）家族的七种不同蛇毒素来选择交叉反应性抗体：磷脂酶 A、长链α-神经毒素和短链α-神经毒素。我们展示了交叉筛选如何增加从噬菌体展示实验中发现交叉反应性单链可变片段（scFv）的机会。此外，我们发现，仅通过分析抗原的序列、结构或表面相似性，很难预测使用交叉筛选发现交叉反应性抗体的可行性。然而，当抗原具有（完全）相同的功能时，这似乎会增加选择交叉反应性抗体的机会，这可能是由于抗原上存在结构相似的基序。
