Rong Yinghui, Chen I-Ling, Larrabee Lance, Sawant Manali S, Fuh Germaine, Koenig Patrick
23andMe, Inc. Therapeutics, 349 Oyster Point Boulevard, South San Francisco, CA 94080, USA.
Antibodies (Basel). 2024 Feb 8;13(1):14. doi: 10.3390/antib13010014.
Bispecific antibodies have gained increasing popularity as therapeutics as they enable novel activities that cannot be achieved with monospecific antibodies. Some of the most popular bispecific formats are molecules in which two Fab arms with different antigen specificities are combined into one IgG-like molecule. One way to produce these bispecific molecules requires the discovery of antibodies against the two antigens of interest that share a common light chain. Here, we present the generation and characterization of a common light chain mouse model, in which the endogenous IGKJ cluster is replaced with a prearranged, modified murine IGKV10-96/IGKJ1 segment. We demonstrate that genetic modification does not impact B-cell development. Upon immunization with ovalbumin, the animals generate an antibody repertoire with VH gene segment usage of a similar diversity to wildtype mice, while the light chain diversity is restricted to antibodies derived from the prearranged IGKV10-96/IGKJ1 germline. We further show that the clonotype diversity of the common light chain immune repertoire matches the diversity of immune repertoire isolated from wildtype mice. Finally, the common light chain anti-ovalbumin antibodies have only slightly lower affinities than antibodies isolated from wildtype mice, demonstrating the suitability of these animals for antibody discovery for bispecific antibody generation.
双特异性抗体作为治疗药物越来越受到欢迎,因为它们能够实现单特异性抗体无法达成的新活性。一些最常见的双特异性形式是将具有不同抗原特异性的两个Fab臂组合成一个IgG样分子的分子。生产这些双特异性分子的一种方法需要发现针对两种感兴趣抗原且共享一条共同轻链的抗体。在此,我们展示了一种共同轻链小鼠模型的构建及表征,其中内源性IGKJ基因簇被一个预先安排的、经过修饰的鼠源IGKV10 - 96/IGKJ1片段所取代。我们证明基因修饰不会影响B细胞发育。在用卵清蛋白免疫后,这些动物产生的抗体库中VH基因片段的使用多样性与野生型小鼠相似,而轻链多样性则局限于源自预先安排的IGKV10 - 96/IGKJ1种系的抗体。我们进一步表明,共同轻链免疫库的克隆型多样性与从野生型小鼠分离的免疫库多样性相匹配。最后,共同轻链抗卵清蛋白抗体的亲和力仅略低于从野生型小鼠分离的抗体,这表明这些动物适用于双特异性抗体生成的抗体发现。