RAMIHM generates fully human monoclonal antibodies by rapid mRNA immunization of humanized mice and BCR-seq

Published: 16 December 2022| Version 3 | DOI: 10.17632/4dcgs8mvzx.3
Ping REN, Peng Lei, Luojia Yang, Kazushi Suzuki, Zhenhao Fang, Paul Renauer, Qianqian Lin, Meizhu Bai, Tongqing Li, Paul Clark, Deryl Klein, Sidi Chen


As a clinical vaccine, lipid nanoparticle (LNP) mRNA demonstrated potent and broad antibody responses, leading to speculation about its potential for antibody discovery. Here, we developed RAMIHM, a highly efficient strategy for developing fully human monoclonal antibodies that employs rapid mRNA immunization of humanized mice followed by single B cell sequencing (scBCR-seq). We immunized humanized transgenic mice with RAMIHM and generated 15 top-ranked clones from peripheral blood, plasma B, and memory B cell populations, demonstrating a high rate of antigen-specificity (93.3%). Two Omicron-specific neutralizing antibodies with high potent and one broad-spectrum neutralizing antibody were discovered. Furthermore, we extended the application of RAMIHM to cancer immunotherapy targets, including a single transmembrane protein CD22 and a multi-transmembrane GPCR target, GPRC5D, which is difficult for traditional protein immunization methods. RAMIHM-scBCR-seq is a broadly applicable platform for the rapid and efficient development of fully human monoclonal antibodies against an assortment of targets.



Yale University Yale Cancer Center


Flow Cytometry