A Phage Display-Identified Peptide Selectively Binds to Kidney Injury Molecule-1 (KIM-1) and Detects KIM-1–Overexpressing Tumors In Vivo
- Author:
Md Enamul HAQUE
1
;
Fatima KHAN
;
Lianhua CHI
;
Smriti GURUNG
;
Sri Murugan Poongkavithai VADEVOO
;
Rang Woon PARK
;
Dong Kyu KIM
;
Sang Kyoon KIM
;
Byungheon LEE
Author Information
- Publication Type:Original Article
- Keywords: In vivo imaging; Kidney injury molecule-1; Peptide; Phage display; Kidney neoplasms
- MeSH: Animals; Bacteriophages; Clone Cells; Enzyme-Linked Immunosorbent Assay; Flow Cytometry; Fluorometry; Kidney Neoplasms; Kidney; Liver; Lung; Mass Screening; Mice; Optical Imaging; Peptide Library; Peptides
- From:Cancer Research and Treatment 2019;51(3):861-875
- CountryRepublic of Korea
- Language:English
- Abstract: PURPOSE: This study was carried out to identify a peptide that selectively binds to kidney injury molecule-1 (KIM-1) by screening a phage-displayed peptide library and to use the peptide for the detection of KIM-1overexpressing tumors in vivo. MATERIALS AND METHODS: Biopanning of a phage-displayed peptide library was performed on KIM-1–coated plates. The binding of phage clones, peptides, and a peptide multimer to the KIM-1 protein and KIM-1–overexpressing and KIM-1–low expressing cells was examined by enzyme-linked immunosorbent assay, fluorometry, and flow cytometry. A biotin-peptide multimer was generated using NeutrAvidin. In vivo homing of the peptide to KIM-1–overexpressing and KIM1–low expressing tumors in mice was examined by whole-body fluorescence imaging. RESULTS: A phage clone displaying the CNWMINKEC peptide showed higher binding affinity to KIM-1 and KIM-1–overexpressing 769-P renal tumor cells compared to other phage clones selected after biopanning. The CNWMINKEC peptide and a NeutrAvidin/biotin-CNWMINKEC multimer selectively bound to KIM-1 over albumin and to KIM-1–overexpressing 769-P cells and A549 lung tumor cells compared to KIM-1–low expressing HEK293 normal cells. Co-localization and competition assays using an anti–KIM-1 antibody demonstrated that the binding of the CNWMINKEC peptide to 769-P cells was specifically mediated by KIM-1. The CNWMINKEC peptide was not cytotoxic to cells and was stable for up to 24 hours in the presence of serum. Whole-body fluorescence imaging demonstrated selective homing of the CNWM-INKEC peptide to KIM-1–overexpressing A498 renal tumor compared to KIM1–low expressing HepG2 liver tumor in mice. CONCLUSION: The CNWMINKEC peptide is a promising probe for in vivo imaging and detection of KIM-1‒overexpressing tumors.
