The phylogenetic distance between chickens and humans accounts for a strong immune response and a broader epitope coverage compared to rodent immunization approaches. Here the authors report the isolation of common light chain (cLC)‐based chicken monoclonal antibodies from an anti‐epidermal growth factor receptor (EGFR) immune library utilizing yeast surface display in combination with yeast biopanning and fluorescence‐activated cell sorting (FACS). For the selection of high‐affinity antibodies, a yeast cell library presenting cLC‐comprising fragment antigen binding (Fab) fragments is panned against hEGFR‐overexpressing A431 cells. The resulting cell–cell‐complexes are sorted by FACS resulting in gradual enrichment of EGFR‐binding Fabs in three sorting rounds. The isolated antibodies share the same light chain and show high specificity for EGFR, resulting in selective binding to A431 cells with notable EC50 values. All identified antibodies show very good aggregation propensity profiles and thermostabilities. Additionally, epitope binning demonstrates that these cLC antibodies cover a broad epitope space. Isolation of antibodies from immunized chickens by yeast cell biopanning makes an addition to the repertoire of methods for antibody library screening, paving the way for the generation of cLC‐based bispecific antibodies against native mammalian receptors.

中文翻译：

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使用酵母生物淘选和荧光激活细胞分选法从免疫鸡中分离常见轻链抗体

与啮齿动物免疫方法相比，鸡与人之间的系统发育距离导致了强烈的免疫反应和更广泛的表位覆盖。在这里，作者报告了利用酵母表面展示结合酵母生物淘选和荧光激活细胞分选（FACS）从抗表皮生长因子受体（EGFR）免疫文库中分离出基于常见轻链（cLC）的鸡单克隆抗体的方法。为了选择高亲和力的抗体，将含有cLC片段抗原结合（Fab）片段的酵母细胞文库针对过表达hEGFR的A431细胞进行淘选。通过FACS对产生的细胞-细胞复合物进行分选，从而在三轮分选中逐渐富集EGFR结合的Fab。分离出的抗体具有相同的轻链，并且对EGFR具有高特异性，导致选择性结合具有显着EC50值的A431细胞。所有鉴定出的抗体都显示出非常好的聚集倾向和热稳定性。此外，抗原决定部位合并证明这些cLC抗体覆盖了广阔的抗原决定部位空间。通过酵母细胞生物淘选从免疫鸡中分离出抗体，为抗体库筛选方法提供了新的内容，为抗天然哺乳动物受体的基于cLC的双特异性抗体的产生铺平了道路。