Department of Genetics, Yale University School of Medicine, New Haven, CT 06520, USA; System Biology Institute, Yale University, West Haven, CT 06520, USA; Center for Cancer Systems Biology, Yale University, West Haven, CT 06520, USA.
Department of Genetics, Yale University School of Medicine, New Haven, CT 06520, USA; System Biology Institute, Yale University, West Haven, CT 06520, USA; Center for Cancer Systems Biology, Yale University, West Haven, CT 06520, USA; Molecular Cell Biology, Genetics, and Development Program, Yale University, New Haven, CT 06520, USA.
Cell Chem Biol. 2023 Jan 19;30(1):85-96.e6. doi: 10.1016/j.chembiol.2022.12.005. Epub 2023 Jan 13.
As a clinical vaccine, lipid nanoparticle (LNP) mRNA has 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 potency 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 G protein-coupled receptor 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.
作为一种临床疫苗，脂质纳米颗粒（LNP）mRNA 已显示出强大而广泛的抗体反应，这引发了人们对其抗体发现潜力的猜测。在这里，我们开发了 RAMIHM，这是一种高效的开发全人源单克隆抗体的策略，它采用了对人源化小鼠进行快速 mRNA 免疫接种，然后进行单个 B 细胞测序（scBCR-seq）。我们用人源化转基因小鼠进行了 RAMIHM 免疫接种，并从外周血、血浆 B 细胞和记忆 B 细胞群中生成了 15 个排名最高的克隆，显示出高抗原特异性（93.3%）。我们发现了两种针对奥密克戎的中和抗体，其具有高效力和一种广谱中和抗体。此外，我们还将 RAMIHM 的应用扩展到癌症免疫治疗靶点，包括单个跨膜蛋白 CD22 和多跨膜 G 蛋白偶联受体靶点 GPRC5D，这些靶点对于传统的蛋白质免疫接种方法来说具有挑战性。RAMIHM-scBCR-seq 是一种广泛适用的平台，可用于快速高效地开发针对各种靶点的全人源单克隆抗体。
