Institute for Organic Chemistry and Biochemistry, Technical University of Darmstadt, Darmstadt, Germany.
Ferring Darmstadt Laboratory, Biologics Technology and Development, Darmstadt, Germany.
Front Immunol. 2020 Dec 23;11:606878. doi: 10.3389/fimmu.2020.606878. eCollection 2020.
Bispecific (BsAb) and biparatopic (BpAb) antibodies emerged as promising formats for therapeutic biologics exhibiting tailor-made functional properties. Over recent years, chicken-derived antibodies have gained traction for diagnostic and therapeutic applications due to their broad epitope coverage and convenience of library generation. Here we report the first generation of a biparatopic common light chain (cLC) chicken-derived antibody by an epitope binning-based screening approach using yeast surface display. The resulting monospecific antibodies target conformational epitopes on domain II or III of the epidermal growth factor receptor (EGFR) with lower double- or single-digit nanomolar affinities, respectively. Furthermore, the domain III targeting variant was shown to interfere with epidermal growth factor (EGF) binding. Utilizing the Knob-into-Hole technology (KiH), a biparatopic antibody with subnanomolar affinity was generated that facilitates clustering of soluble and cell-bound EGFR and displayed enhanced antibody-dependent cell-mediated cytotoxicity (ADCC) compared to the parental antibodies. This strategy for generating cLC-based biparatopic antibodies from immunized chickens may pave the way for their further development in therapeutic settings.
双特异性 (BsAb) 和双价 (BpAb) 抗体作为治疗性生物制剂的有前途的形式出现，具有定制的功能特性。近年来，由于具有广泛的表位覆盖范围和方便的文库生成，鸡源性抗体在诊断和治疗应用中引起了关注。在这里，我们报告了第一代基于表位分组的筛选方法通过酵母表面展示产生的双价通用轻链 (cLC) 鸡源性抗体。由此产生的单特异性抗体针对表皮生长因子受体 (EGFR) 的结构域 II 或 III 上的构象表位，亲和力分别为低至双位数或个位数纳摩尔。此外，靶向域 III 的变体被证明可以干扰表皮生长因子 (EGF) 的结合。利用 Knob-into-Hole 技术 (KiH)，生成了具有亚纳摩尔亲和力的双价抗体，可促进可溶性和细胞结合的 EGFR 的聚集，并显示出比亲本抗体更高的抗体依赖性细胞介导的细胞毒性 (ADCC)。这种从免疫鸡中生成基于 cLC 的双价抗体的策略可能为它们在治疗环境中的进一步发展铺平道路。
