抗体体外多样化数据集

碱基编辑器 (BE) 是用于精确校正或多样化突变的新兴工具。本数据集提供了一种新方法来重建体细胞超突变 (SHM) 以产生高亲和力抗体，该抗体通常从抗原免疫后的动物模型或合成组合文库中筛选出来。通过比较基因编辑后相同基因位点的体细胞突变，我们将CRISPR系统与脱氨酶偶联，并对脱氨酶进行蛋白质工程设计，进而更新了多样化的碱基编辑器 (DBE) 以生成 SHM。 DBE 中使用的脱氨酶保留了其固有的核苷酸偏好性并对其靶向热点处的胞苷进行突变。融合AID 的 DBE 与体内生理状态的AID蛋白具有相同的靶向热点，而带有其它脱氨酶的 DBE靶向同一DNA 底物将产生不同的突变图谱。最后，DBE 在抗体展示系统中的应用实现了抗体体外亲和力成熟。我们的研究结果解释了 DBE 靶向编辑机制和一种新型抗体工程化方法。

Base editors (BEs) are emerging tools for precise correction or diversification of mutations. This dataset presents a novel method to reconstruct somatic hypermutation (SHM) for generating high-affinity antibodies, which are typically screened from antigen-immunized animal models or synthetic combinatorial libraries. By comparing somatic mutations at identical genetic loci following gene editing, we coupled the CRISPR system with deaminases and conducted protein engineering on the deaminases, thereby updating diversified base editors (DBEs) to generate SHM. The deaminases employed in DBEs retain their inherent nucleotide preferences and mutate cytidines at their targeting hotspots. AID-fused DBEs exhibit the same targeting hotspots as physiologically active AID proteins in vivo, while DBEs paired with other deaminases targeting the same DNA substrate yield distinct mutation profiles. Finally, the application of DBEs in antibody display systems enables in vitro affinity maturation of antibodies. Our findings elucidate the targeted editing mechanisms of DBEs and a novel antibody engineering strategy.

搜集汇总

背景与挑战

背景概述

该数据集聚焦于利用碱基编辑器（BE）作为新兴工具，通过重建体细胞超突变（SHM）来生成高亲和力抗体。它介绍了将CRISPR系统与脱氨酶偶联并更新多样化碱基编辑器（DBE）的方法，应用于抗体展示系统以实现抗体体外亲和力成熟。