Zhang Yangde, Chen Jiji, Zhang Yanqiong, Hu Zhiyuan, Hu Duosha, Pan Yifeng, Ou Sheng, Liu Gang, Yin Xiang, Zhao Jingfeng, Ren Lifeng, Wang Jiwei
National Key Laboratory of Nanobiological Technology, Changsha, Hunan, China.
J Biomol Screen. 2007 Apr;12(3):429-35. doi: 10.1177/1087057106299164. Epub 2007 Mar 1.
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.
在临床试验中，肿瘤靶向治疗可能是治愈恶性肿瘤的一种有效方法。噬菌体展示是一种分子多样性技术，可使大量肽或蛋白质在丝状噬菌体表面呈现，用于各种应用。在本研究中，我们报告了利用噬菌体展示生成与结肠癌组织结合的肽库。为此，我们开发了一种筛选方案，包括对结肠癌细胞（SW480）进行3轮体外阳性淘选，以及利用噬菌体展示-7肽库对正常人肠上皮细胞进行2轮体外消减筛选。经过几轮淘选，噬菌体滴度和回收效率均显著提高。通过基于细胞的酶联免疫吸附测定、免疫荧光、体内结合测定、免疫细胞化学染色和免疫组织化学染色，证明肽CP15（VHLGYAT）是靶向肿瘤细胞（SW480和HT29细胞）和肿瘤组织，但不靶向正常人肠上皮细胞和对照结肠组织的最有效肽。这些研究表明，肽CP15可能是开发有用的结肠肿瘤诊断和靶向给药剂的一个有前景的先导候选物。
