Wash-free and selective imaging of epithelial cell adhesion molecule (EpCAM) expressing cells with fluorogenic peptide ligands

Tara Bahadur K C; Kanako Suga; Takashi Isoshima; Toshiro Aigaki; Yoshihiro Ito; Kiyotaka Shiba; Takanori Uzawa

doi:10.1016/j.bbrc.2018.04.059

Wash-free and selective imaging of epithelial cell adhesion molecule (EpCAM) expressing cells with fluorogenic peptide ligands

Biochem Biophys Res Commun. 2018 Jun 2;500(2):283-287. doi: 10.1016/j.bbrc.2018.04.059.

Authors

Tara Bahadur K C¹, Kanako Suga², Takashi Isoshima³, Toshiro Aigaki⁴, Yoshihiro Ito⁵, Kiyotaka Shiba⁶, Takanori Uzawa⁷

Affiliations

¹ 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.
² Division of Protein Engineering, Cancer Institute, Japanese Foundation for Cancer Research, 3-8-31, Ariake, Koto, Tokyo 135-8550, Japan.
³ Nano Medical Engineering Laboratory, RIKEN, 2-1 Hirosawa, Wako, Saitama 351-0198, Japan.
⁴ Graduate School of Biological Science, Tokyo Metropolitan University, 1-1 Minami-Osawa, Tokyo 192-0397, Japan; Nano Medical Engineering Laboratory, RIKEN, 2-1 Hirosawa, Wako, Saitama 351-0198, Japan.
⁵ 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; Nano Medical Engineering Laboratory, RIKEN, 2-1 Hirosawa, Wako, Saitama 351-0198, Japan.
⁶ Division of Protein Engineering, Cancer Institute, Japanese Foundation for Cancer Research, 3-8-31, Ariake, Koto, Tokyo 135-8550, Japan. Electronic address: kshiba@jfcr.or.jp.
⁷ Emergent Bioengineering Materials Research Team, RIKEN Center for Emergent Matter Science, 2-1 Hirosawa, Wako, Saitama 351-0198, Japan; Nano Medical Engineering Laboratory, RIKEN, 2-1 Hirosawa, Wako, Saitama 351-0198, Japan. Electronic address: tuzawa@riken.jp.

PMID: 29660346
DOI: 10.1016/j.bbrc.2018.04.059

Abstract

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.

Keywords: Aptamer; CTC diagnosis; Circulating tumor cell; Fluorescence imaging; Liquid biopsy for cancer.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Amino Acid Sequence
Cell Line, Tumor
Cell Membrane / metabolism
Epithelial Cell Adhesion Molecule / chemistry
Epithelial Cell Adhesion Molecule / metabolism*
Fluorescent Dyes / metabolism*
Humans
Ligands
Molecular Imaging / methods*
Peptides / chemistry
Peptides / metabolism*
Protein Binding

Substances

Epithelial Cell Adhesion Molecule
Fluorescent Dyes
Ligands
Peptides