一种靶向EGFR/VEGRFR2的双特异性抗体的体外抗肿瘤活性研究

【作者】 徐孟怀； 金海珍； 张娟； 王旻；

【机构】 中国药科大学生命科学与技术学院；

【摘要】 背景:VEGFR2和EGFR信号通路相互作用,同时抑制这两条通路的抗肿瘤效果优于仅抑制单一信号通路。本实验室构建了同时靶向VEGFR2和EGFR通路的双特异性双链抗体Db,以期获得增强的抗肿瘤活性。方法:利用SPR技术、流式细胞术、MTT、划痕损伤修复及凋亡分析实验对Db进行抗肿瘤活性研究。结果:Db能与重组人EGFR和VEGRFR2蛋白高亲和力结合,并能结合到高表达EGFR的A431细胞和高表达VEGFR2的HUVEC细胞表面,抑制A431细胞/HUVEC细胞的增殖及迁移,通过生理性程序促进细胞凋亡。结论:靶向EGFR/VEGRFR2的双特异性抗体Db保持亲本抗体的生物物理学活性,具有肿瘤活性,可用于进一步抗肿瘤研究。更多还原

【Abstract】 Background:EGFR-VEGFR pathway cross-talk underlines a potential joint targeting for antibody-based therapy.We previously constructed a novel a bispecific diabody(Db) targeting both EGFR and VEGFR2 for enhanced anti-tumor activity.Methods:Db was firstly characterized by surface plasmon resonance(SPR) and flow cytometry.Then Db was assayed with its anti-tumor activity through MTT assay,wound healing assay and apoptosis assay.Results:The Db bound to both recombinant human EGFR/VEGFR protein and EGFR over-expressing A431 cells and VEGFR2 over-expressing HUVECs.Db demonstrated both dose and time dependent inhibition on the proliferation and migration of A431 cells and HUVECs,and proved to be potent apoptosis inducer in both cell lines in a dose dependent manner.Conclusion:Db was successfully constructed,expressed and purified with the individual scFvs retaining their functional properties.The Db displays favorable biophysical properties for biopharmaceutical development,therefore could potentially be used as co-targeted therapy for EGFR and VEGFR2 over-expressing tumors.更多还原