RT Journal
A1 Wang, Yuhong
A1 Guo, Yuefeng
A1 Lu, Qiuyu
A1 Liu, Xinyi
A1 Xu, Haoqi
A1 Chen, Jiayi
A1 Pi, Ruiqi
A1 Yuan, Shaopeng
A1 Yang, Ziting
A1 Lu, Rusen
A1 Meng, Fei-Long
A1 Gan, Tingting
A1 Hu, Jiazhi
T1 Restoring the potency of neutralizing antibody via guided hypermutation with hyper-antibody editor
JF Genome Research 
JO Genome Research 
YR 2026 
FD March 19 
DO 10.1101/gr.281396.125 
SP gr.281396.125 
UL http://genome.cshlp.org/content/early/2026/03/18/gr.281396.125.abstract 
AB 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.