Emergent Bioengineering Materials Research Team, RIKEN Center for Emergent Matter Science, 2-1 Hirosawa, Wako, Saitama 351-0198, Japan; Graduate School of Biological Science, Tokyo Metropolitan University, 1-1 Minami-Osawa, Tokyo 192-0397, Japan.
Division of Protein Engineering, Cancer Institute, Japanese Foundation for Cancer Research, 3-8-31, Ariake, Koto, Tokyo 135-8550, Japan.
Biochem Biophys Res Commun. 2018 Jun 2;500(2):283-287. doi: 10.1016/j.bbrc.2018.04.059.
Detection of the cells expressing an epithelial cell adhesion molecule (EpCAM) is a crucial step to identify circulating tumor cells (CTCs) from blood. To detect the EpCAM, we here designed and synthesized a series of fluorogenic peptides. Specifically, we functionalized an EpCAM-binding peptide, Ep114, by replacing its amino acids to an aminophenylalanine that was modified with environmentally sensitive 7-nitro-2,1,3-benzoxadiazole (NBD-amPhe). Among six synthesized peptides, we have found that two peptides, Q4X and V6X (X represents NBD-amPhe), retain the Ep114's binding ability and specifically mark EpCAM-expressing cells by just adding these peptides to the cultivation medium. Our wash-free, fluorogenic peptide ligands would boost the development of next generation devices for CTC diagnoses.
检测表达上皮细胞黏附分子（EpCAM）的细胞是从血液中鉴定循环肿瘤细胞（CTC）的关键步骤。为了检测 EpCAM，我们在这里设计并合成了一系列荧光肽。具体来说，我们通过用环境敏感的 7-硝基-2,1,3-苯并恶二唑（NBD-amPhe）修饰的氨基苯丙氨酸取代 EpCAM 结合肽 Ep114 的氨基酸，对其进行了功能化。在合成的六种肽中，我们发现两种肽，Q4X 和 V6X（X 代表 NBD-amPhe），保留了 Ep114 的结合能力，只需将这些肽添加到培养介质中，就可以特异性地标记表达 EpCAM 的细胞。我们的无洗涤、荧光肽配体将推动下一代 CTC 诊断设备的发展。
