Léxane Fournier, Lukas Pekar, Birgitta Leuthner, Harald Kolmar, Lars Toleikis, Stefan Becker
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引用次数: 0
摘要
在这项工作中,我们报告了通过结合驼科动物免疫和荧光激活细胞分选(FACS)发现的强效抗表皮生长因子受体(EGFR)异构重链抗体。在免疫和酵母表面展示文库构建之后,通过引入标记的表皮生长因子受体 Fc 融合蛋白作为 FACS 的附加标准,获得了异构克隆。这种分选方法鉴定出了 11 种重链抗体,它们与表皮生长因子受体的结合不会与正交配体表皮生长因子受体竞争。这些抗体与表达表皮生长因子受体的三阴性乳腺癌细胞系结合,亲和力在皮摩尔到纳摩尔范围内。这些源自骆驼的抗体还通过调节表皮生长因子受体对表皮生长因子受体的亲和力而表现出有趣的特性。此外,它们还能抑制表皮生长因子受体诱导的下游信号通路。特别是,我们发现一种克隆抗体在抑制PI3K/AKT和MAPK/ERK通路方面比已获批准的阻断抗体西妥昔单抗更有效。我们的研究结果表明,异构抗体可能是一种潜在的新治疗方式。
Discovery of potent allosteric antibodies inhibiting EGFR.
In this work, we report the discovery of potent anti-epidermal growth factor receptor (EGFR) allosteric heavy-chain antibodies by combining camelid immunization and fluorescence-activated cell sorting (FACS). After immunization and yeast surface display library construction, allosteric clones were obtained by introducing the labeled EGF Fc fusion protein as an additional criterion for FACS. This sorting method enabled the identification of 11 heavy-chain antibodies that did not compete with the orthosteric ligand EGF for the binding to EGFR. These antibodies bind to a triple-negative breast cancer cell line expressing EGFR with affinities in the picomolar to nanomolar range. Those camelid-derived antibodies also exhibit interesting properties by modulating EGFR affinity for EGF. Moreover, they are also able to inhibit EGF-induced downstream signaling pathways. In particular, we identified one clone that is more potent than the approved blocking antibody cetuximab in inhibiting both PI3K/AKT and MAPK/ERK pathways. Our results suggest that allosteric antibodies may be potential new modalities for therapeutics.
期刊介绍:
mAbs is a multi-disciplinary journal dedicated to the art and science of antibody research and development. The journal has a strong scientific and medical focus, but also strives to serve a broader readership. The articles are thus of interest to scientists, clinical researchers, and physicians, as well as the wider mAb community, including our readers involved in technology transfer, legal issues, investment, strategic planning and the regulation of therapeutics.