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Journal of Biomolecular Screening
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Article

Panning and Identification of a Colon Tumor Binding Peptide from a Phage Display Peptide Library

Yangde Zhang1, Jiji Chen1, Yanqiong Zhang1, Zhiyuan Hu1, Duosha Hu2, Yifeng Pan3, Sheng Ou1, Gang Liu2, Xiang Yin1, Jingfeng Zhao1, Lifeng Ren1, Jiwei Wang1*

1 National Key Laboratory of Nanobiological Technology, Changsha, Hunan, China.
2 Cancer Research Institute, Central South University, Changsha, Hunan, China.
3 National Hepatobiliary & Enteric Surgery Research Center, Central South University, Changsha, Hunan, China.

* To whom correspondence should be addressed. E-mail: wangjiwei{at}xysm.net.


  Abstract

Tumor-targeting therapy can be an efficacious way to cure a malignant tumor in clinical trials. Phage display is a molecular diversity technology that allows the presentation of a large number of peptides or proteins on the surface of filamentous phage for various applications. In this study, we report on using phage display to generate peptide libraries that bind to colon cancer tissues. To accomplish this, we developed a screening protocol that contained 3 rounds of in vitro positive panning on colon cancer cells (SW480) and 2 rounds of subtractive screening in vitro on normal human intestinal epithelial cells with a phage display-7 peptide library. After several rounds of panning, both phage titer and recovery efficiency were significantly improved. Through a cell-based enzyme-linked immunosorbent assay, immunofluorescence, in vivo binding assay, immunocytochemical staining, and immunohistochemical staining, peptide CP15 (VHLGYAT) was demonstrated to be the most effective peptide in targeting tumor cells (SW480 and HT29 cells) and tumor tissues but not the normal human intestinal epithelial cells and control colon tissue. These studies suggest that peptide CP15 may be a promising lead candidate in the development of a useful colon tumor diagnostic and targeted drug delivery agent.

Key Words: colon cancer, phage display, targeting peptides, SW480 cells, HT29 cells

First published on March 1, 2007, doi:10.1177/1087057106299164

Journal of Biomolecular Screening 2007;12:429.

A more recent version of this article appeared on April 1, 2007

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