Counting of Enzymatically Amplified Affinity Reactions in Hydrogel Particle-templated Drops

Overview

Journal Lab Chip

Publisher Royal Society of Chemistry

Specialties Biotechnology
Chemistry

Date 2021 Aug 11

PMID 34378611

Citations 8

Authors

Yilian Wang

Vishwesh Shah

Angela Lu

Ella Pachler

Brian Cheng

Dino Di Carlo

Affiliations

Soon will be listed here.

Abstract

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.

Citing Articles

Lab on a Particle Technologies.

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Fluid dynamics alters liquid-liquid phase separation in confined aqueous two-phase systems.

Hester E, Carney S, Shah V, Arnheim A, Patel B, Di Carlo D Proc Natl Acad Sci U S A. 2023; 120(49):e2306467120.

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Digital Cascade Assays for ADP- or ATP-Producing Enzymes Using a Femtoliter Reactor Array Device.

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Hydrogel-Based Multi-enzymatic System for Biosynthesis.

Wu H, Zheng B Adv Biochem Eng Biotechnol. 2023; 186:51-76.

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