Karami Elmira, Mesbahi Moghaddam Maryam, Behdani Mahdi, Kazemi-Lomedasht Fatemeh
Biotechnology Department, Biotechnology Research Center, Pasteur Institute of Iran, Tehran, Iran.
Department of Biology, Shahed University, Tehran, Iran.
Prep Biochem Biotechnol. 2023;53(5):523-531. doi: 10.1080/10826068.2022.2111583. Epub 2022 Aug 19.
Neuropilin-1 (NRP-1) is a non-tyrosine kinase receptor and when overexpressed, leads to angiogenesis. High expression of NRP-1 has been observed in various cancers. Unique characteristic of nanobodies (small size, high affinity and stability, and ease production) make them potential therapeutic tools. Oligoclonal nanobodies which detect multiple functional epitopes on the target antigen could be potential tools for inhibition of cancer resistance problems due to escape variant of tumor cells. In this study, oligoclonal anti-NRP-1 nanobodies were selected from camel immune library and their binding activities as well as functionality were evaluated. Anti-NRP-1 nanobodies were expressed in an host, and purified using nickel affinity chromatography. The effect of each individual and oligoclonal nanobodies on human endothelial cells were evaluated by MTT, Tube formation, and migration assay as well. Results showed that oligoclonal anti-NRP-1 nanobodies detected different epitopes of NRP-1 antigen and inhibited angiogenesis of human endothelial cells better than each individual nanobody. Results indicate promising oligoclonal anti-NRP-1 nanobodies for inhibition of angiogenesis.
神经纤毛蛋白-1（NRP-1）是一种非酪氨酸激酶受体，过度表达时会导致血管生成。在各种癌症中均观察到NRP-1的高表达。纳米抗体的独特特性（体积小、亲和力和稳定性高以及易于生产）使其成为潜在的治疗工具。能够检测靶抗原上多个功能表位的寡克隆纳米抗体可能是抑制因肿瘤细胞逃逸变体导致的癌症耐药问题的潜在工具。在本研究中，从骆驼免疫文库中筛选出寡克隆抗NRP-1纳米抗体，并对其结合活性和功能进行了评估。抗NRP-1纳米抗体在宿主中表达，并通过镍亲和层析进行纯化。还通过MTT、管形成和迁移试验评估了单个和寡克隆纳米抗体对人内皮细胞的作用。结果表明，寡克隆抗NRP-1纳米抗体可检测到NRP-1抗原的不同表位，并且比单个纳米抗体更有效地抑制人内皮细胞的血管生成。结果表明，寡克隆抗NRP-1纳米抗体在抑制血管生成方面具有良好前景。
