High-Affinity Internalizing Human scFv-Fc Antibody for Targeting FGFR1-Overexpressing Lung Cancer
High-Affinity Internalizing Human scFv-Fc Antibody for Targeting FGFR1-Overexpressing Lung Cancer
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DOI：
10.1158/1541-7786.mcr-16-0136
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发表时间：
2017-08-01
影响因子：
5.2
通讯作者：
Otlewski, Jacek
中科院分区：
文献类型：
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作者：
Sokolowska-Wedzina, Aleksandra;Chodaczek, Grzegorz;Otlewski, Jacek
使用针对肿瘤相关抗原的抗体靶向输送抗癌药物是当前免疫肿瘤学研究中最重要的方法之一。成纤维细胞生长因子受体1(FGFR1)是一种针对吸烟相关肺癌的高频靶向癌基因，存在于超过20%的肺鳞癌病例中。本报告描述了以scFv-Fc形式高效地靶向FGFR1的有效的全人抗体片段的产生。用噬菌体展示技术筛选抗FGFR1胞外区(IIIc)的高亲和力单链抗体片段。采用酶免疫分析(ELISA)和表面等离子体共振(SPR)分析进行抗体筛选和鉴定。最好的粘合剂(命名为D2)被克隆到Diabody和FC融合格式。所有D2抗体均与FGFR1有较高的亲和力，其解离常数分别为18nmol/L(ScFvD2)、0.82nmol/L(ScFvD2)和0.59nmol/L(scFvD2-Fc)。竞争分析表明，与其他FGFR家族成员相比，scFvD2对FGFR1具有很好的选择性，并且即使在其天然配体FGF2存在的情况下也能与FGFR1结合。共聚焦显微镜显示，scFvD2-Fc被一组过表达FGFR1的细胞系特异性地迅速内化。最后，证实了scFvD2-Fc介导的细胞毒性有效载荷特异性地传递到含有致癌FGFR1基因扩增的肺癌细胞中。(C)2017年AACR。
Targeted delivery of anticancer drugs using antibodies specific for tumor-associated antigens represents one of themost important approaches in current immuno-oncology research. Fibroblast growth factor receptor 1 (FGFR1) has been demonstrated to be a high-frequency targetable oncogene specific for smoking-associated lung cancers, present in over 20% of lung squamous cell carcinoma cases. This report describes the generation of a potent, fully human antibody fragment in scFv-Fc format efficiently targeting FGFR1. Antibody phage display was used to select high-affinity scFv antibody fragments against the extracellular domain of FGFR1(IIIc). Enzyme immunoassay (ELISA) and surface plasmon resonance (SPR) analysis were used for antibody screening and characterization. The best binder (named D2) was cloned to diabody and Fc fusion formats. All D2 antibodies demonstrated high affinity for FGFR1 with dissociation constants of 18 nmol/L (scFvD2), 0.82 nmol/L (scFvD2 diabody), and 0.59 nmol/L (scFvD2-Fc). scFvD2 was found to be exquisitely selective for FGFR1 versus other FGFR family members and bound FGFR1 even in the presence of its natural ligand FGF2, as shown by competitive analysis. Confocal microscopy revealed that scFvD2-Fc was specifically and rapidly internalized by a panel of cell lines overexpressing FGFR1. Finally, it was demonstrated that scFvD2-Fc mediated specific delivery of a cytotoxic payload into lung cancer cells harboring oncogenic FGFR1 gene amplifications. (C) 2017 AACR.