Labeling of VEGFR1D2 through oxime ligation
Labeling of VEGFR1D2 through oxime ligation
复制标题
DOI：
10.1016/j.bioorg.2019.103160
复制
发表时间：
2019-10-01
影响因子：
5.1
通讯作者：
D'Andrea, Luca Domenico
中科院分区：
文献类型：
--
作者：
De Rosa, Lucia;Di Stasi, Rossella;D'Andrea, Luca Domenico
我们报告了一个有用的协议标记的第二个结构域的血管内皮生长因子受体1（VEGFR1D2），一个小的蛋白质配体能够结合VEGF，血管生成的主要调节。我们开发了一种生物缀合策略，其基于使用肟连接反应将VEGFR1D2的醛衍生物缀合至具有烷氧基胺官能团的分子探针。我们应用合成方案来制备VEGFR1D2的生物素化缀合物，并且我们证明该生物缀合物保留其以高亲和力特异性结合其天然配体VEGF的能力。生物素化的VEGFR1D2可用于检测和定量VEGF以用于诊断目的，以及用于筛选靶向VEGF的新分子以用于治疗应用的工具。标记协议是通用的，并可以扩展到不同的分子探针，如荧光团，螯合剂或多聚体支架，为VEGF靶向提供生物医学平台。
We reported an useful protocol for the labeling of the second domain of the Vascular Endothelial Growth Factor Receptor 1 (VEGFR1D2), a small protein ligand able to bind VEGF, the main regulator of angiogenesis. We developed a bioconjugation strategy based on the use of oxime-ligation reaction conjugating an aldehyde derivative of the VEGFR1D2 to a molecular probe harboring an alkoxyamine functional group. We applied the synthetic protocol to prepare a biotinylated conjugate of VEGFR1D2 and we demonstrate that the bioconjugate retains its ability to specifically bind its natural ligand, VEGF, with high affinity. The biotinylated VEGFR1D2 could be useful to detect and quantify VEGF for diagnostic purposes as well as a tool for the screening of new molecules targeting VEGFRs for therapeutic applications. The labeling protocol is versatile and can be extended to different molecular probes, such as fluorophores, chelators or multimeric scaffolds, affording a biomedical platform for VEGF targeting.