Tang Ping, Li Li, Zhou Yan, Shen Cong-Cong, Kang Yu-Huan, Yao Yu-Qin, Yi Cheng, Gou Lan-Tu, Yang Jin-Liang
State Key Laboratory of Biotherapy and Cancer Center, West China Hospital, West China Medical School, Sichuan University, Chengdu, People's Republic of China.
Department of Biochemistry and Molecular Biology, West China School of Preclinical and Forensic Medicine, Sichuan University, Chengdu, People's Republic of China.
Biotechnol Appl Biochem. 2014 Jul-Aug;61(4):376-84. doi: 10.1002/bab.1187. Epub 2014 Mar 20.
Bispecific antibody (BsAb) has been proved to be a very effective antitumor approach because of its distinctive advantages of immune-mediated cytotoxicity. To enhance the ability to recruit and activate T lymphocytes for tumor-specific killing, we constructed and prepared a recombinant human single-chain Fv bispecific antibody (BsAb), named VEGFR1/CD3 BsAb, targeting VEGFR1 and CD3. The VEGFR1/CD3 BsAb was expressed in CHO-K1 cells and purified by Ni-NTA affinity chromatography. The CD3 and VEGFR1-binding activity of VEGFR1/CD3 BsAb was confirmed by flow cytometry. T lymphocyte activation and proliferation induced by VEGFR1/CD3 BsAb were also demonstrated in vitro. Notably, our VEGFR1/CD3 BsAb presented a powerful and specific killing effect against VEGFR1-positive human breast cancer cell MDA-MB-231 and MDA-MB-435 through activating T lymphocyte at very low concentrations, indicating that it will be a valuable antibody drug for treatment of VEGFR1-positive cancers in the future.
双特异性抗体（BsAb）因其免疫介导的细胞毒性的独特优势，已被证明是一种非常有效的抗肿瘤方法。为了增强招募和激活T淋巴细胞进行肿瘤特异性杀伤的能力，我们构建并制备了一种重组人单链Fv双特异性抗体（BsAb），命名为VEGFR1/CD3 BsAb，靶向VEGFR1和CD3。VEGFR1/CD3 BsAb在CHO-K1细胞中表达，并通过Ni-NTA亲和层析进行纯化。通过流式细胞术证实了VEGFR1/CD3 BsAb的CD3和VEGFR1结合活性。体外实验也证明了VEGFR1/CD3 BsAb可诱导T淋巴细胞活化和增殖。值得注意的是，我们的VEGFR1/CD3 BsAb在极低浓度下通过激活T淋巴细胞，对VEGFR1阳性的人乳腺癌细胞MDA-MB-231和MDA-MB-435呈现出强大而特异的杀伤作用，表明它未来将成为治疗VEGFR1阳性癌症的一种有价值的抗体药物。
