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A Phage Display-Identified Peptide Selectively Binds to Kidney Injury Molecule-1 (KIM-1) and Detects KIM-1–Overexpressing Tumors In Vivo / Journal of the Korean Cancer Association, 대한암학회지
Cancer Research and Treatment ; : 861-875, 2019.
Article in English | WPRIM | ID: wpr-763192
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.
Subject(s)

Full text: Available Index: WPRIM (Western Pacific) Main subject: Peptides / Bacteriophages / Enzyme-Linked Immunosorbent Assay / Mass Screening / Clone Cells / Peptide Library / Optical Imaging / Flow Cytometry / Fluorometry / Kidney Type of study: Screening study Limits: Animals Language: English Journal: Cancer Research and Treatment Year: 2019 Type: Article

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Full text: Available Index: WPRIM (Western Pacific) Main subject: Peptides / Bacteriophages / Enzyme-Linked Immunosorbent Assay / Mass Screening / Clone Cells / Peptide Library / Optical Imaging / Flow Cytometry / Fluorometry / Kidney Type of study: Screening study Limits: Animals Language: English Journal: Cancer Research and Treatment Year: 2019 Type: Article