Biopanning and rapid analysis of selective interactive ligands
Biopanning and rapid analysis of selective interactive ligands
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DOI：
10.1038/nm1101-1249
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发表时间：
2001-11-01
期刊：
影响因子：
82.9
通讯作者：
Arap, W
中科院分区：
文献类型：
--
作者：
Giordano, RJ;Cardó-Vila, M;Arap, W
在这里，我们介绍了一种从噬菌体库中筛选、选择和分选细胞表面结合肽的新方法。选择性相互作用配体（称为 BRASIL）的生物淘选和快速分析基于差速离心，其中在水性上相中与噬菌体一起孵育的细胞悬浮液通过不混溶的有机下相进行离心。这种单步有机相分离比目前依赖洗涤步骤或有限稀释的方法更快、更灵敏、更特异。作为原理验证，我们筛选了用血管内皮生长因子 (VEGF) 刺激的人内皮细胞，并构建了 VEGF 家族基于肽的配体受体图谱。接下来，我们验证了 PQPRPL 基序作为一种新型嵌合配体模拟物，可特异性结合 VEGF 受体 1 和神经毡蛋白 1。 BRASIL 可能证明自己是一种探测靶细胞表面的优越方法，具有广泛的潜在应用。
Here we introduce a new approach for the screening, selection and sorting of cell-surface-binding peptides from phage libraries. Biopanning and rapid analysis of selective interactive ligands (termed BRASIL) is based on differential centrifugation in which a cell suspension incubated with phage in an aqueous upper phase is centrifuged through a non-miscible organic lower phase. This single-step organic phase separation is faster, more sensitive and more specific than current methods that rely on washing steps or limiting dilution. As a proof-of-principle, we screened human endothelial cells stimulated with vascular endothelial growth factor (VEGF) and constructed a peptide-based ligand-receptor map of the VEGF family. Next, we validated the motif PQPRPL as a novel chimeric ligand mimic that binds specifically to VEGF receptor-1 and to neuropilin-1. BRASIL may prove itself a superior method for probing target cell surfaces with a broad range of potential applications.