The predominant approach for antibody generation remains animal immunization, which can yield exceptionally selective and potent antibody clones owing to the powerful evolutionary process of somatic hypermutation. However, animal immunization is inherently slow, not always accessible and poorly compatible with many antigens. Here, we describe ‘autonomous hypermutation yeast surface display’ (AHEAD), a synthetic recombinant antibody generation technology that imitates somatic hypermutation inside engineered yeast. By encoding antibody fragments on an error-prone orthogonal DNA replication system, surface-displayed antibody repertoires continuously mutate through simple cycles of yeast culturing and enrichment for antigen binding to produce high-affinity clones in as little as two weeks. We applied AHEAD to generate potent nanobodies against the SARS-CoV-2 S glycoprotein, a G-protein-coupled receptor and other targets, offering a template for streamlined antibody generation at large.

产生抗体的主要方法仍然是动物免疫，由于体细胞超突变的强大进化过程，它可以产生特别选择性和有效的抗体克隆。然而，动物免疫本质上是缓慢的，并不总是容易获得并且与许多抗原的相容性差。在这里，我们描述了“自主超突变酵母表面展示”（AHEAD），这是一种合成重组抗体生成技术，可模拟工程酵母内部的体细胞超突变。通过在容易出错的正交 DNA 复制系统上编码抗体片段，表面展示的抗体库通过酵母培养和抗原结合富集的简单循环持续突变，从而在短短两周内产生高亲和力克隆。