Counting of enzymatically amplified affinity reactions in hydrogel particle-templated drops,Lab on a Chip

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Counting of enzymatically amplified affinity reactions in hydrogel particle-templated drops
Lab on a Chip ( IF 6.1 ) Pub Date : 2021-08-04 , DOI: 10.1039/d1lc00344e
Yilian Wang ₁ , Vishwesh Shah ₁ , Angela Lu ₁ , Ella Pachler ₁ , Brian Cheng ₁ , Dino Di Carlo _{1,

2}

Affiliation

Counting of numerous compartmentalized enzymatic reactions underlies quantitative and high sensitivity immunodiagnostic assays. However, digital enzyme-linked immunosorbent assays (ELISA) require specialized instruments which have slowed adoption in research and clinical labs. We present a lab-on-a-particle solution to digital counting of thousands of single enzymatic reactions. Hydrogel particles are used to bind enzymes and template the formation of droplets that compartmentalize reactions with simple pipetting steps. These hydrogel particles can be made at a high throughput, stored, and used during the assay to create ∼500 000 compartments within 2 minutes. These particles can also be dried and rehydrated with sample, amplifying the sensitivity of the assay by driving affinity interactions on the hydrogel surface. We demonstrate digital counting of β-galactosidase enzyme at a femtomolar detection limit with a dynamic range of 3 orders of magnitude using standard benchtop equipment and experiment techniques. This approach can faciliate the development of digital ELISAs with reduced need for specialized microfluidic devices, instruments, or imaging systems.

中文翻译：

水凝胶颗粒模板滴中酶促放大亲和反应的计数

对众多区室化酶反应的计数是定量和高灵敏度免疫诊断测定的基础。然而，数字酶联免疫吸附测定 (ELISA) 需要专用仪器，这阻碍了研究和临床实验室的采用。我们提出了一种粒子实验室解决方案，可对数千个单一酶反应进行数字计数。水凝胶颗粒用于结合酶并模板形成液滴，通过简单的移液步骤将反应区分开。这些水凝胶颗粒可以高通量制造、储存并在分析过程中使用，在 2 分钟内创建约 500 000 个隔室。这些颗粒还可以干燥并与样品一起再水合，通过驱动水凝胶表面上的亲和相互作用来增强测定的灵敏度。我们使用标准台式设备和实验技术，演示了在飞摩尔检测限和 3 个数量级动态范围内对 β-半乳糖苷酶进行数字计数。这种方法可以促进数字 ELISA 的开发，减少对专用微流体设备、仪器或成像系统的需求。

更新日期：2021-08-10

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