Share Email Print

Journal of Biomedical Optics • Open Access

Cy5.5-labeled Affibody molecule for near-infrared fluorescent optical imaging of epidermal growth factor receptor positive tumors
Author(s): Zheng Miao; Gang Ren; Hongguang Liu; Lei Jiang; Zhen Cheng

Paper Abstract

Affibody protein is an engineered protein scaffold with a three-helical bundle structure. Affibody molecules of small size (7 kD) have great potential for targeting overexpressed cancer biomarkers in vivo. To develop an Affibody-based molecular probe for in vivo optical imaging of epidermal growth factor receptor (EGFR) positive tumors, an anti-EGFR Affibody molecule, Ac-Cys-ZEGFR:1907 (7 kD), is site-specifically conjugated with a near-IR fluorescence dye, Cy5.5-mono-maleimide. Using fluorescent microscopy, the binding specificity of the probe Cy5.5-ZEGFR:1907 is checked by a high-EGFR-expressing A431 cell and low-EGFR-expressing MCF7 cells. The binding affinity of Cy5.5-ZEGFR:1907 (KD) to EGFR is 43.6±8.4 nM, as determined by flow cytometry. For an in vivo imaging study, the probe shows fast tumor targeting and good tumor contrast as early as 0.5 h postinjection (p.i.) for A431 tumors, while MCF7 tumors are barely visible. An ex vivo imaging study also demonstrates that Cy5.5-ZEGFR:1907 has high tumor, liver, and kidney uptakes at 24 h p.i.. In conclusion, Cy5.5-ZEGFR:1907 shows good affinity and high specificity to the EGFR. There is rapid achievement of good tumor-to-normal-tissue contrasts of Cy5.5-ZEGFR:1907, thus demonstrating its potential for EGFR-targeted molecular imaging of cancers.

Paper Details

Date Published: 1 May 2010
PDF: 7 pages
J. Biomed. Opt. 15(3) 036007 doi: 10.1117/1.3432738
Published in: Journal of Biomedical Optics Volume 15, Issue 3
Show Author Affiliations
Zheng Miao, Stanford Univ. (United States)
Gang Ren, Stanford Univ. (United States)
Hongguang Liu, Stanford Univ. (United States)
Lei Jiang, Stanford Univ. (United States)
Zhen Cheng, Stanford Univ. (United States)

© SPIE. Terms of Use
Back to Top