Department of Chemistry , Tanta University , Tanta 31527 , Egypt.
Department of Pharmacology & Physiology , George Washington University , Washington , D.C. 20037 , United States.
Anal Chem. 2018 Jun 19;90(12):7569-7577. doi: 10.1021/acs.analchem.8b01198. Epub 2018 May 31.
We report here the fabrication and validation of a novel 3D-printed, automated immunoarray to detect multiple proteins with ultralow detection limits. This low cost, miniature immunoarray employs electrochemiluminescent (ECL) detection measured with a CCD camera and employs touch-screen control of a micropump to facilitate automated use. The miniaturized array features prefilled reservoirs to deliver sample and reagents to a paper-thin pyrolytic graphite microwell detection chip to complete sandwich immunoassays. The detection chip achieves high sensitivity by using single-wall carbon nanotube-antibody conjugates in the microwells and employing massively labeled antibody-decorated RuBPY-silica nanoparticles to generate ECL. The total cost of an array is $0.65, and an eight-protein assay can be done in duplicate for $0.14 per protein with limits of detection (LOD) as low as 78-110 fg mL in diluted serum. The electronic control system costs $210 in components. Utility of the automated immunoarray was demonstrated by detecting an eight-protein prostate cancer biomarker panel in human serum samples in 25 min. The system is well suited to future clinical and point-of-care diagnostic testing and could be used in resource-limited environments.
我们在此报告一种新型的 3D 打印自动化免疫阵列的制造和验证，该阵列可用于检测多种具有超低检测限的蛋白质。这种低成本、微型免疫阵列采用电化学发光（ECL）检测，通过 CCD 相机进行测量，并采用触摸屏控制微泵，以方便自动化使用。微型阵列配备有预填充的储液器，可将样品和试剂输送到薄如纸的热解石墨微井检测芯片上，以完成三明治免疫分析。该检测芯片通过在微井中使用单壁碳纳米管-抗体缀合物，并采用大量标记的抗体修饰的 RuBPY-二氧化硅纳米颗粒来产生 ECL，从而实现高灵敏度。阵列的总成本为 0.65 美元，每个阵列可以进行两次八蛋白检测，每个蛋白的检测费用为 0.14 美元，在稀释的血清中的检测限（LOD）低至 78-110 fg mL。电子控制系统的组件成本为 210 美元。自动化免疫阵列的实用性通过在 25 分钟内检测人血清样本中的八种前列腺癌生物标志物进行了验证。该系统非常适合未来的临床和即时诊断测试，并且可以在资源有限的环境中使用。
