使用抗 EGF 抗体从自动展示的 Fv 抗体库中筛选超热生长因子受体 (EGFR) 抑制剂。 Fv抗体文库表达于大肠杆菌外膜，对应免疫球蛋白G的重链VH区。通过定点随机化表达的VH区（11个氨基酸残基）的CDR3区构建文库。诱变。使用抗EGF抗体作为筛选探针，从Fv抗体库中筛选具有抗体结合亲和力的氨基酸序列（CDR3区）。这些氨基酸序列被认为与EGF具有相似的化学性质，可以与EGFR结合。从Fv抗体库中筛选出两个带有针对EGFR的Fv抗体的自展示克隆，并且筛选的Fv抗体表达为可溶性蛋白。使用SPR生物传感器估计结合亲和力(KD)，并使用EGFR介导的信号通路中的蛋白质的蛋白质印迹分析观察表达的Fv抗体对PANC-1胰腺肿瘤细胞和T98G胶质母细胞瘤细胞的抑制活性。观察到 PANC-1 和 T98G 细胞的活力因表达的 Fv 抗体的抑制活性而降低。最后，通过分子对接模拟分析了 Fv 抗体与 EGFR 之间的相互作用。

Inhibitors of the epithermal growth factor receptor (EGFR) were screened from an autodisplayed Fv-antibody library using an anti-EGF antibody. The Fv-antibody library was expressed on the outer membrane of Escherichia coli, which corresponds to the heavy chain VH region of immunoglobulin G. The library was constructed by randomizing the CDR3 region of expressed VH regions (11 amino acid residues) by site-directed mutagenesis. Using an anti-EGF antibody as a screening probe, amino acid sequences (CDR3 region) with antibody binding affinity were screened from the Fv-antibody library. These amino acid sequences were considered to have similar chemical properties to EGF, which can bind to EGFR. Two autodisplayed clones with Fv-antibodies against EGFR were screened from the Fv-antibody library, and the screened Fv-antibodies were expressed as soluble proteins. The binding affinity (KD) was estimated using an SPR biosensor, and the inhibitory activity of expressed Fv-antibodies was observed for PANC-1 pancreatic tumor cells and T98G glioblastoma cells using Western blot analysis of proteins in the EGFR-mediated signaling pathway. The viability of PANC-1 and T98G cells was observed to decrease via the inhibitory activity of expressed Fv-antibodies. Finally, interactions between Fv-antibodies and EGFR were analyzed by using molecular docking simulations.