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Single Molecule Protein Detection with Attomolar Sensitivity Using Droplet Digital Enzyme-Linked Immunosorbent Assay.
ACS Nano ( IF 17.1 ) Pub Date : 2020-06-26 , DOI: 10.1021/acsnano.0c02378
Limor Cohen 1, 2, 3 , Naiwen Cui 4 , Yamei Cai 4 , Padric M Garden 1, 2 , Xiang Li 1, 2 , David A Weitz 1, 4 , David R Walt 1, 2, 3
Affiliation  

Many proteins are present at low concentrations in biological samples, and therefore, techniques for ultrasensitive protein detection are necessary. To overcome challenges with sensitivity, the digital enzyme-linked immunosorbent assay (ELISA) was developed, which is 1000× more sensitive than conventional ELISA and allows sub-femtomolar protein detection. However, this sensitivity is still not sufficient to measure many proteins in various biological samples, thereby limiting our ability to detect and discover biomarkers. To overcome this limitation, we developed droplet digital ELISA (ddELISA), a simple approach for detecting low protein levels using digital ELISA and droplet microfluidics. ddELISA achieves maximal sensitivity by improving the sampling efficiency and counting more target molecules. ddELISA can detect proteins in the low attomolar range and is up to 25-fold more sensitive than digital ELISA using Single Molecule Arrays (Simoa), the current gold standard tool for ultrasensitive protein detection. Using ddELISA, we measured the LINE1/ORF1 protein, a potential cancer biomarker that has not been previously measured in serum. Additionally, due to the simplicity of our device design, ddELISA is promising for point-of-care applications. Thus, ddELISA will facilitate the discovery of biomarkers that have never been measured before for various clinical applications.

中文翻译:

使用液滴数字酶联免疫吸附测定法检测具有大分子敏感性的单分子蛋白质。

许多蛋白质以低浓度存在于生物样品中,因此,超灵敏蛋白质检测技术必不可少。为了克服灵敏度方面的挑战,开发了数字酶联免疫吸附测定(ELISA),其灵敏度比常规ELISA高1000倍,并可以检测亚飞摩尔蛋白。但是,这种敏感性仍然不足以测量各种生物样品中的许多蛋白质,从而限制了我们检测和发现生物标志物的能力。为克服此限制,我们开发了液滴数字ELISA(ddELISA),这是一种使用数字ELISA和液滴微流控技术检测低蛋白水平的简单方法。ddELISA通过提高采样效率和对更多目标分子进行计数来实现最大灵敏度。ddELISA可以检测低摩尔浓度的蛋白质,并且比使用单分子阵列(Simoa)的数字ELISA灵敏度高25倍,Simoa是目前用于检测超灵敏蛋白质的金标准工具。使用ddELISA,我们测量了LINE1 / ORF1蛋白,这是一种潜在的癌症生物标记物,以前从未在血清中测量过。此外,由于我们设备设计的简单性,ddELISA在即时医疗应用中很有前途。因此,ddELISA将促进发现各种临床应用之前从未测量过的生物标志物。以前从未在血清中测量过的潜在癌症生物标志物。此外,由于我们设备设计的简单性,ddELISA在即时医疗应用中很有前途。因此,ddELISA将促进发现各种临床应用之前从未测量过的生物标志物。以前从未在血清中测量过的潜在癌症生物标志物。此外,由于我们设备设计的简单性,ddELISA在即时医疗应用中很有前途。因此,ddELISA将促进发现各种临床应用之前从未测量过的生物标志物。
更新日期:2020-08-25
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