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基于设计的分支肽的复杂生物介质中超低污染癌细胞定量的电化学适体传感器。

Electrochemical Aptasensor for Ultralow Fouling Cancer Cell Quantification in Complex Biological Media Based on Designed Branched Peptides.

机构信息

Key Laboratory of Optic-electric Sensing and Analytical Chemistry for Life Science, MOE; Shandong Key Laboratory of Biochemical Analysis; Key Laboratory of Analytical Chemistry for Life Science in Universities of Shandong; College of Chemistry and Molecular Engineering . Qingdao University of Science and Technology , Qingdao 266042 , PR China.

Department of Chemistry , University of Oxford , Oxford OX1 3QZ , United Kingdom.

出版信息

Anal Chem. 2019 Jul 2;91(13):8334-8340. doi: 10.1021/acs.analchem.9b01129. Epub 2019 Jun 10.

Abstract

The rapid, convenient, and selective assaying of clinical targets directly in complex biological media brings with it the potential to revolutionize diagnostics. One major hurdle to impact is retention of selectivity and a tight control of nonspecific surface interactions or biofouling. We report herein, the construction of an antifouling interface through the covalent attachment of designed branched zwitterionic peptides onto electrodeposited polyaniline film. The antifouling capability of the designed branched peptide significantly outperforms that of the commonly used PEG and linear peptides. The interfaces modified with branched peptides are exceptionally effective in reducing a nonspecific protein and cell adsorption, as verified by electrochemical and fluorescent characterization. The derived sensors with mucin1 protein (MUC1) aptamer as the recognition element detect MUC1-positive MCF-7 breast cancer cells in human serum with high sensitivity and selectivity. The linear response range of the cytosensor for the MCF-7 cell is from 50 to 10 cells/mL, with a limit of detection as low as 20 cells/mL. More importantly, the assaying performances remain unchanged in human serum owing to the presence of branched antifouling peptide, indicating feasibility of the cytosensor for practical cancer cell quantification in complex samples.

摘要

直接在复杂生物介质中快速、便捷、选择性地分析临床靶标有可能带来诊断学的革命。但要产生实际影响,主要障碍之一是保留选择性和严格控制非特异性表面相互作用或生物污垢。本文报道了通过将设计的支链两性离子肽共价附着到电沉积聚苯胺膜上来构建抗污染界面。与常用的 PEG 和线性肽相比,设计的支链肽具有出色的抗污染能力。经电化学和荧光特性验证,支链肽修饰的界面能非常有效地减少非特异性蛋白质和细胞的吸附。衍生的基于粘蛋白 1 蛋白(MUC1)适体的传感器作为识别元件,以高灵敏度和选择性检测人血清中的 MUC1 阳性 MCF-7 乳腺癌细胞。该细胞传感器对 MCF-7 细胞的线性响应范围为 50 至 10 个细胞/mL,检测限低至 20 个细胞/mL。更重要的是,由于存在支链抗污肽,在人血清中检测性能保持不变,表明该细胞传感器在复杂样品中用于实际癌细胞定量分析的可行性。

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