An integrated microfluidic system for screening of peptides specific to colon cancer cells and colon cancer stem cells using the phage display technology

Yu-Jui Che, Huei-Wen Wu, Lein-Yu Hung, Hwan-You Chang, Kuan Wang, Gwo-Bin Lee
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Abstract

Reagents binding specifically to target molecules are essential tools for clinical diagnosis and targeted therapy. Screening of target cell-surface specific affinity reagents with bench-top methods has some drawbacks, including time-consuming, labor-intensive and requirement of large-scale instrument. Microfluidic platforms may overcome these drawbacks because they could automate and complete the screening process within a shorter period of time. Phage display is a promising technology in selection of cell-surface specific peptides. In recent years, a subpopulation of tumor cells named cancer stem cells is believed to be the tumorigenic cells and closely associated with metastasis. A specific peptide that can recognize and differentiate cancer stem cells from the rest of cancer cell population is therefore useful for early diagnosis and targeted therapy. In this study, selection of M13 phage displayed peptides that bind with colon cancer cells and colon cancer stem cells using an integrated microfluidic system was successfully demonstrated. Compared with the traditional methods, the total selection process was shortened to 36 hours while traditional method needs almost a month. More importantly, the screening process can be automated and performed on a single microfluidic chip. The developed technique may be promising for early diagnosis of cancer and target therapeutics.
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利用噬菌体展示技术筛选结肠癌细胞和结肠癌干细胞特异性肽的集成微流控系统
特异性结合靶分子的试剂是临床诊断和靶向治疗的重要工具。利用台式方法筛选靶细胞表面特异性亲和试剂存在耗时、劳动强度大、需要大型仪器等缺点。微流控平台可以克服这些缺点,因为它们可以在更短的时间内自动化并完成筛选过程。噬菌体展示是一种很有前途的细胞表面特异性肽选择技术。近年来,一种被称为肿瘤干细胞的肿瘤细胞亚群被认为是致瘤细胞,并与转移密切相关。因此,一种能够识别和区分癌症干细胞与其他癌细胞群的特异性肽对于早期诊断和靶向治疗是有用的。在本研究中,利用集成的微流体系统成功地选择了M13噬菌体显示的与结肠癌细胞和结肠癌干细胞结合的肽。与传统方法相比,整个选择过程缩短至36小时,而传统方法需要近一个月的时间。更重要的是,筛选过程可以自动化,并在单个微流控芯片上进行。该技术有望用于癌症的早期诊断和靶向治疗。
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