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基于水凝胶的悬浮阵列用于生物标志物检测,使用辣根过氧化物酶介导的银沉淀。

Hydrogel-based suspension array for biomarker detection using horseradish peroxidase-mediated silver precipitation.

机构信息

Bioengineering Program, Department of Mechanical Engineering, University of Michigan, Dearborn, 4901 Evergreen Road, Dearborn, MI 48128, United States.

Bioengineering Program, Department of Mechanical Engineering, University of Michigan, Dearborn, 4901 Evergreen Road, Dearborn, MI 48128, United States.

出版信息

Anal Chim Acta. 2018 Jan 25;999:132-138. doi: 10.1016/j.aca.2017.10.033. Epub 2017 Oct 30.

DOI:10.1016/j.aca.2017.10.033
PMID:29254564
Abstract

Advances in medical diagnostics and personalized therapy require robust, sensitive yet cost-effective diagnostic tools for rapid measurement of biomolecules including proteins in body fluids. State-of-the-art technologies are complex and rely on expensive or custom made detection system, and therefore, cannot be readily adapted for point-of-care (POC) analysis. The development of a novel detection platform, which leverages horseradish peroxidase (HRP)-mediated silver precipitation within antibody immobilized porosity tuned poly (ethylene) glycol diacrylate (PEGDA) hydrogel microparticles with the operational advantages of suspension arrays for sensitive quantification of biomarkers, is described. In this study, vascular endothelial growth factor (VEGF) has been used as a model protein. The silver deposition corresponded to the concentration of VEGF in solution. The detection limit of 5.2 ± 1.0 pg/mL and assay time of 2 h highlights that this assay exceeds the conventional technologies in terms of sensitivity and speed. The practical applicability of the hydrogel microparticle based detection system has been established by demonstrating the ability of the system to quantify the production of VEGF by highly aggressive (MDA-MB-231) and non-aggressive (MCF-7) breast cancer cells. The reliance on simple instrument for quantification of clinically relevant markers bolsters the adaptability of the detection platform/method in POC settings.

摘要

医学诊断和个性化治疗的进步需要强大、敏感且具有成本效益的诊断工具,以便快速测量生物分子,包括体液中的蛋白质。最先进的技术复杂且依赖昂贵或定制的检测系统,因此不能轻易适应即时护理(POC)分析。本研究中,血管内皮生长因子(VEGF)被用作模型蛋白。银沉积与溶液中 VEGF 的浓度相对应。检测限为 5.2 ± 1.0 pg/mL,检测时间为 2 小时,这表明该检测方法在灵敏度和速度方面优于传统技术。通过证明该系统能够定量测定高度侵袭性(MDA-MB-231)和非侵袭性(MCF-7)乳腺癌细胞产生的 VEGF,确立了基于水凝胶微球的检测系统的实际适用性。该检测平台/方法对临床相关标志物的定量依赖于简单的仪器,增强了其在 POC 环境下的适应性。

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