Behdani Mahdi, Zeinali Sirous, Karimipour Morteza, Khanahmad Hossein, Asadzadeh Nader, Azadmanesh Kayhan, Seyed Negar, Baniahmad Seyed Farzad, Anbouhi Mahdi Habibi
Department of Molecular Medicine, Pasteur Institute of Iran, Tehran, Iran.
Adv Biomed Res. 2012;1:34. doi: 10.4103/2277-9175.100126. Epub 2012 Aug 28.
Antibodies and their derivative fragments have long been used as tools in a variety of applications, in fundamental research work, biotechnology, diagnosis, and therapy. Camels produce single heavy-chain antibodies (VHH) in addition to usual antibodies. These minimal-sized binders are very robust and bind the antigen with high affinity in a monomeric state. Vascular endothelial growth factor recepror-2 (VEGFR2) is an important tumor-associated receptor that blockade of its signaling can lead to the inhibition of neovascularization and tumor metastasis. Here, we describe the construction, expression, and purification VEGFR2-specific Diabody. Two variable fragments of a same camel anti-VEGFR2 antibody were linked together by the upper hinge segment of antibody to make a diabody. We showed the ability of diabody to recognition of VEGFR2 on the cell surface by FACS. Diabodies can be produced in the low-cost prokaryotic expression system, so they are suitable molecules for diagnostic and therapeutic issues.
抗体及其衍生片段长期以来一直被用作各种应用中的工具，包括基础研究工作、生物技术、诊断和治疗。骆驼除了产生常规抗体外，还产生单链重抗体（VHH）。这些最小尺寸的结合物非常稳定，以单体状态与抗原高亲和力结合。血管内皮生长因子受体-2（VEGFR2）是一种重要的肿瘤相关受体，阻断其信号传导可导致新血管形成和肿瘤转移的抑制。在这里，我们描述了VEGFR2特异性双体的构建、表达和纯化。同一骆驼抗VEGFR2抗体的两个可变片段通过抗体的上铰链段连接在一起，形成一个双体。我们通过荧光激活细胞分选术（FACS）展示了双体识别细胞表面VEGFR2的能力。双体可以在低成本的原核表达系统中产生，因此它们是适用于诊断和治疗问题的分子。
