Venom and Biotherapeutics Molecules Laboratory, Biotechnology Department, Biotechnology Research Center, Pasteur Institute of Iran, Tehran, Islamic Republic of Iran.
Immunopharmacol Immunotoxicol. 2021 Apr;43(2):230-238. doi: 10.1080/08923973.2021.1888114. Epub 2021 Mar 3.
Neuropilin-1 (NRP-1) regulates a range of physiological and pathological processes, including angiogenesis. Targeting of NRP1 is considered a significant approach in cancer therapy. In the present study, a novel antiNRP1 immunotoxin (αNRP1 IT) was developed by genetic fusion of a single domain (VHH) anti-NRP-1 antibody fragment to a truncated diphtheria toxin. The αNRP1 IT was expressed into bacterial cells as an inclusion body (IB). Expression of αNRP1 IT was confirmed by SDS-PAGE and western blotting. Recombinant αNRP1 IT was purified using nickel affinity chromatography. Toxicity and antiangiogenesis effect of αNRP1 IT was investigated both and . Results showed that αNRP1 IT significantly reduced the viability of human umbilical vein endothelial cell line (HUVEC) ( < .05). The αNRP1 IT significantly inhibited tube formation of HUVEC cells ( < .001). Furthermore, αNRP1 IT inhibited angiogenesis in Chick Chorioallantoic Membrane (CAM) Assay. These data suggest the potential of αNRP1 IT as a novel therapeutic in targeted cancer therapy.
神经纤毛蛋白-1（NRP-1）调节多种生理和病理过程，包括血管生成。靶向 NRP1 被认为是癌症治疗的一种重要方法。在本研究中，通过将单域（VHH）抗 NRP-1 抗体片段与截短的白喉毒素基因融合，开发了一种新型抗 NRP1 免疫毒素（αNRP1 IT）。αNRP1 IT 作为包涵体（IB）在细菌细胞中表达。通过 SDS-PAGE 和 Western blot 确认 αNRP1 IT 的表达。使用镍亲和层析法纯化重组 αNRP1 IT。研究了 αNRP1 IT 的毒性和抗血管生成作用。结果表明，αNRP1 IT 显著降低人脐静脉内皮细胞系（HUVEC）的活力（<0.05）。αNRP1 IT 显著抑制 HUVEC 细胞的管形成（<0.001）。此外，αNRP1 IT 抑制鸡胚绒毛尿囊膜（CAM）分析中的血管生成。这些数据表明，αNRP1 IT 作为一种新型治疗药物具有在靶向癌症治疗中的潜力。
