Desai Alec A, Zupancic Jennifer M, Trzeciakiewicz Hanna, Gerson Julia E, DuBois Kelly N, Skinner Mary E, Sharkey Lisa M, McArthur Nikki, Ferris Sean P, Bhatt Nemil N, Makowski Emily K, Smith Matthew D, Chen Hongwei, Huang Jie, Jerez Cynthia, Kuo Yun-Huai, Kane Ravi S, Kanaan Nicholas M, Paulson Henry L, Tessier Peter M
Department of Chemical Engineering, University of Michigan, Ann Arbor, MI, USA.
Biointerfaces Institute, University of Michigan, Ann Arbor, MI, USA.
Nat Chem Biol. 2025 Apr 29. doi: 10.1038/s41589-025-01881-9.
Antibodies that recognize insoluble antigens, such as amyloid fibrils associated with neurodegenerative disorders, are important for research, diagnostic and therapeutic applications. However, these types of antibodies are difficult to generate, typically require animal immunization and also commonly require humanization in the case of therapeutic applications. Here we report a methodology for generating high-quality, fully human, conformation-specific antibodies against amyloid fibrils using a published human nonimmune library, yeast-surface display and quantitative fluorescence-activated cell sorting. Notably, this approach enables the isolation of conformation-specific antibodies against tau fibrils (Alzheimer's disease) and α-synuclein fibrils (Parkinson's disease) with combinations of high affinity, high conformational specificity and, in some cases, low off-target binding that rival or exceed those of clinical-stage antibodies specific for tau (zagotenemab) and α-synuclein (cinpanemab). This approach is expected to simplify the generation of conformation-specific antibodies against diverse protein aggregates and other insoluble antigens.
识别不溶性抗原（如与神经退行性疾病相关的淀粉样纤维）的抗体对于研究、诊断和治疗应用都很重要。然而，这类抗体难以产生，通常需要动物免疫，而且在治疗应用中通常还需要人源化。在此，我们报告一种使用已发表的人类非免疫文库、酵母表面展示和定量荧光激活细胞分选来生成针对淀粉样纤维的高质量、完全人源、构象特异性抗体的方法。值得注意的是，这种方法能够分离出针对tau纤维（阿尔茨海默病）和α-突触核蛋白纤维（帕金森病）的构象特异性抗体，其具有高亲和力、高构象特异性，在某些情况下还具有低脱靶结合的组合，可与或超过针对tau（扎戈替奈单抗）和α-突触核蛋白（辛帕奈单抗）的临床阶段抗体。预计这种方法将简化针对多种蛋白质聚集体和其他不溶性抗原的构象特异性抗体的产生。
