Department of Cell Biology, Pittsburgh, PA, USA; Department Computational and Systems Biology, Pittsburgh, PA, USA; University of Pittsburgh-Carnegie Mellon University Program in Computational Biology, Pittsburgh, PA, USA.
Department of Cell Biology, Pittsburgh, PA, USA.
Structure. 2022 Mar 3;30(3):418-429.e3. doi: 10.1016/j.str.2021.11.006. Epub 2021 Dec 10.
Nanobodies (Nbs) have emerged as a promising class of biologics. Despite having marked physicochemical properties, Nbs are derived from camelids and may require humanization to improve translational potentials. By systematically analyzing the sequence and structural properties of Nbs, we found substantial framework diversities and revealed the key differences between Nbs and human immunoglobulin G antibodies. We identified conserved residues that may contribute to enhanced solubility, structural stability, and antigen binding, providing insights into Nb humanization. Based on big data analysis, we developed "Llamanade," an open-source software to facilitate rational humanization of Nbs. Using sequence as input, Llamanade can rapidly extract sequence features, model structures, and optimize solutions to humanize Nbs. Finally, we used Llamanade to successfully humanize a cohort of structurally diverse and potent SARS-CoV-2 neutralizing Nbs. Llamanade is freely available and will be easily accessible on a server to support the development of therapeutic Nbs into safe and effective trials.
纳米抗体（Nbs）已经成为一类有前途的生物制剂。尽管具有显著的物理化学特性，但 Nbs 来源于骆驼科动物，可能需要进行人源化改造以提高转化潜能。通过系统分析 Nbs 的序列和结构特性，我们发现了大量的框架多样性，并揭示了 Nbs 与人类免疫球蛋白 G 抗体之间的关键差异。我们鉴定了可能有助于提高溶解度、结构稳定性和抗原结合的保守残基，为 Nb 人源化提供了见解。基于大数据分析，我们开发了“Llamanade”，这是一款开源软件，可促进 Nbs 的合理化人源化。使用序列作为输入，Llamanade 可以快速提取序列特征、建模结构，并优化 Nbs 人源化的解决方案。最后，我们使用 Llamanade 成功地对一组结构多样且有效的 SARS-CoV-2 中和 Nbs 进行了人源化改造。Llamanade 是免费提供的，并将在服务器上轻松访问，以支持将治疗性 Nbs 开发为安全有效的试验。
