Bispecific antibodies have recently gained major interest as they allow novel mechanisms-of-action and/or therapeutic applications that cannot be achieved using conventional IgG-based antibodies. A major issue in engineering IgG-based bispecific antibodies has been to enable the correct association of heavy and light chains resulting in correct assembly of the desired bispecific antibody in sufficient yield. Various approaches have been described during recent years to tackle this challenge. We have developed the so-called CrossMab technology that enforces correct light chain association based on the domain crossover of immunoglobulin domains in the Fab region of the bispecific antibody. This versatile technology allows the generation of different bispecific antibody formats including asymmetric heterodimeric monovalent 1 + 1 bispecific antibodies and asymmetric heterodimeric bispecific antibodies with 2 + 1 valency in combination with approaches enabling Fc-hetermodimerization like knob-into-hole technology as well as the generation of tetravalent symmetric bispecific antibodies with 2 + 2 valency, also known as Tandem-Fab based IgG antibodies, using processes suitable for the large scale production of therapeutic bispecific antibodies. Notably, as of now, at least eight different bispecific antibodies using CrossMab technology entered clinical development, and additional CrossMabs are in late preclinical development. This review provides a summary of the status and progress with the engineering and generation of CrossMab technology based bispecific antibodies as well as their therapeutic application.

中文翻译：

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使用 CrossMab 技术设计治疗性双特异性抗体

双特异性抗体最近引起了人们的极大兴趣，因为它们允许使用传统的基于 IgG 的抗体无法实现的新的作用机制和/或治疗应用。工程化基于 IgG 的双特异性抗体的一个主要问题是使重链和轻链正确结合，从而以足够的产量正确组装所需的双特异性抗体。近年来，已经描述了各种方法来应对这一挑战。我们开发了所谓的 CrossMab 技术，该技术基于双特异性抗体 Fab 区中免疫球蛋白域的域交叉，强制执行正确的轻链关联。这种多功能技术允许生成不同的双特异性抗体形式，包括不对称异二聚体单价 1 + 1 双特异性抗体和具有 2 + 1 价的不对称异源二聚体双特异性抗体，结合实现 Fc 异源二聚化的方法，如旋钮入孔技术以及生成使用适合大规模生产治疗性双特异性抗体的工艺制备具有 2 + 2 价的四价对称双特异性抗体，也称为基于串联 Fab 的 IgG 抗体。值得注意的是，截至目前，至少有八种不同的使用 CrossMab 技术的双特异性抗体进入了临床开发，还有更多的 CrossMab 处于临床前后期开发阶段。