Institute of Immunology, University Medical Center Hamburg-Eppendorf, Hamburg, Germany.
Center for Molecular Neurobiology Hamburg, University Medical Center Hamburg-Eppendorf, Hamburg, Germany.
Nat Commun. 2024 Jun 3;15(1):4728. doi: 10.1038/s41467-024-48735-x.
Due to their exceptional solubility and stability, nanobodies have emerged as powerful building blocks for research tools and therapeutics. However, their generation in llamas is cumbersome and costly. Here, by inserting an engineered llama immunoglobulin heavy chain (IgH) locus into IgH-deficient mice, we generate a transgenic mouse line, which we refer to as 'LamaMouse'. We demonstrate that LamaMice solely express llama IgH molecules without association to Igκ or λ light chains. Immunization of LamaMice with AAV8, the receptor-binding domain of the SARS-CoV-2 spike protein, IgE, IgG2c, and CLEC9A enabled us to readily select respective target-specific nanobodies using classical hybridoma and phage display technologies, single B cell screening, and direct cloning of the nanobody-repertoire into a mammalian expression vector. Our work shows that the LamaMouse represents a flexible and broadly applicable platform for a facilitated selection of target-specific nanobodies.
由于其出色的溶解性和稳定性，纳米抗体已成为研究工具和治疗药物的强大构建模块。然而，在美洲驼中生成纳米抗体既繁琐又昂贵。在这里，我们通过将工程化的美洲驼免疫球蛋白重链（IgH）基因座插入到 IgH 缺陷型小鼠中，生成了一种转基因小鼠品系，我们称之为“LamaMouse”。我们证明 LamaMouse 仅表达不与 Igκ 或 λ 轻链结合的 llama IgH 分子。用 AAV8（SARS-CoV-2 刺突蛋白的受体结合域）、IgE、IgG2c 和 CLEC9A 免疫 LamaMouse，使我们能够使用经典的杂交瘤和噬菌体展示技术、单个 B 细胞筛选以及直接将纳米抗体库克隆到哺乳动物表达载体中，轻松选择各自的靶特异性纳米抗体。我们的工作表明，LamaMouse 代表了一种灵活且广泛适用的平台，可方便地选择靶特异性纳米抗体。
