@article{Wang19032026,
author = {Wang, Yuhong and Guo, Yuefeng and Lu, Qiuyu and Liu, Xinyi and Xu, Haoqi and Chen, Jiayi and Pi, Ruiqi and Yuan, Shaopeng and Yang, Ziting and Lu, Rusen and Meng, Fei-Long and Gan, Tingting and Hu, Jiazhi}, 
title = {Restoring the potency of neutralizing antibody via guided hypermutation with hyper-antibody editor},
year = {2026}, 
doi = {10.1101/gr.281396.125}, 
elocation-id = {gr.281396.125}, 
abstract ={Somatic hypermutation (SHM) drives antibody affinity maturation in B cells. By mimicking this process, guided hypermutation (GHM) tools employing CRISPR systems and activation-induced cytidine deaminase (AID) have advanced antibody development. However, GHM-induced mutations in cultured cells exhibit mutation patterns distinct from those observed in natural antibody diversification following in vivo affinity selection. To address this, we engineer a hyper-antibody editor, HAE1, by integrating cytidine and adenine deaminases with a nicked, PAMless Cas9 variant, SpRY, to closely resemble the mutation spectrum of natural SHM. Moreover, to streamline mutation, selection, and validation within the same cells, we develop a dual-expression system in HEK293F cells that allows simultaneous expression of both transmembrane and secreted full-length antibodies. Using this system, we apply HAE1 to the SARS-CoV-2 neutralizing antibody CV07-209 and restore the antibody's binding affinity and neutralization potency against Omicron variants, specifically BA.1, including at least one mutation beyond the reach of current GHM tools. HAE1 thus provides a versatile, high-throughput strategy for expediting antibody evolution, presenting significant potential for therapeutic antibody development and protein engineering.}, 
URL = {http://genome.cshlp.org/content/early/2026/03/18/gr.281396.125.abstract}, 
eprint = {http://genome.cshlp.org/content/early/2026/03/18/gr.281396.125.full.pdf+html}, 
journal = {Genome Research} 
}