High specificity accompanied with the ability to recruit immune cells has made recombinant therapeutic antibodies an integral part of drug development. Here we present a generic approach to generate two novel IgG-derived antibody formats that are based on a modification of the CrossMab technology. MoAbs harbor two heavy chains (HCs) resulting in one binding entity and one fragment crystallizable region (Fc), whereas DuoMabs are composed of four HCs harboring two binding entities and two Fc regions linked at a disulfide-bridged hinge. The latter bivalent format is characterized by avidity-enhanced target cell binding while simultaneously increasing the ‘Fc-load’ on the surface. DuoMabs were shown to be producible in high yield and purity and bind to surface cells with affinities comparable to IgGs. The increased Fc load directed at the surface of target cells by DuoMabs modulates their antibody-dependent cell-mediated cytotoxicity competency toward target cells, making them attractive for applications that require or are modulated by FcR interactions.

高特异性以及招募免疫细胞的能力使重组治疗抗体成为药物开发不可或缺的一部分。在这里，我们提出了一种通用方法来生成两种新型 IgG 衍生抗体形式，这些抗体形式基于 CrossMab 技术的修改。MoAb 包含两条重链 (HC)，从而形成一个结合实体和一个片段可结晶区 (Fc)，而 DuoMab 由四个 HC 组成，其中包含两个结合实体和两个通过二硫桥铰链连接的 Fc 区。后一种二价形式的特点是亲和力增强的靶细胞结合，同时增加表面的“Fc 负载”。DuoMab 被证明可以高产率和高纯度生产，并以与 IgG 相当的亲和力与表面细胞结合。DuoMabs 增加针对靶细胞表面的 Fc 负载，调节其对靶细胞的抗体依赖性细胞介导的细胞毒性能力，使其对需要 FcR 相互作用或受 FcR 相互作用调节的应用具有吸引力。