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Related Concept Videos

Enzyme-Linked Immunosorbent Assay01:33

Enzyme-Linked Immunosorbent Assay

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In 1971, Peter Perlman and Eva Engvall developed an Enzyme-linked immunosorbent assay (ELISA or EIA). ELISA differs from western blot in that the assays are conducted in microtiter plates or in vivo rather than on an absorbent membrane.
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Related Experiment Video

Updated: Feb 5, 2026

Development and Validation of an Ultrasensitive Single Molecule Array Digital Enzyme-linked Immunosorbent Assay for Human Interferon-&#945;
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Digital enzyme assay using attoliter droplet array.

Takao Ono1, Takanori Ichiki, Hiroyuki Noji

  • 1Department of Applied Chemistry, Graduate School of Engineering, The University of Tokyo, Japan. hnoji@appchem.t.u-tokyo.ac.jp.

The Analyst
|September 18, 2018
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Summary
This summary is machine-generated.

This study introduces nanocell technology for highly sensitive digital enzyme assays. Nanocells enable oil-free, air-sealed reactions, significantly improving fluorescent product accumulation for biomolecule quantification.

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Area of Science:

  • Biochemistry
  • Analytical Chemistry
  • Biotechnology

Background:

  • Digital enzyme assays quantify biomolecules at low concentrations using droplet arrays.
  • Multiplex assays are challenging due to varying optimal buffer conditions and fluorogenic compound properties.

Purpose of the Study:

  • To develop a novel nanocell platform for enhanced digital enzyme assays.
  • To overcome limitations of existing droplet-based assays, including non-optimal enzyme conditions and fluorophore leakage.

Main Methods:

  • Utilized nanometer-sized droplets (200 aL) termed 'nanocells' for digital enzyme assays.
  • Implemented an oil-free sealing method using airflow for nanocell reactors.
  • Performed dual digital enzyme assays with β-galactosidase and alkaline phosphatase (ALP).

Main Results:

  • Nanocells enhanced fluorescent product accumulation by 100x compared to micron-sized reactors.
  • Achieved enzyme catalysis at rates comparable to oil-sealed reactors using air-sealed nanocells.
  • Successfully detected ALP molecules even under non-optimal pH conditions (pH 7.4).

Conclusions:

  • Nanocell technology significantly expands the applicability of digital bioassays for enzymes under non-optimal conditions or with low turnover rates.
  • The air-sealing method broadens the scope of usable fluorescent dyes, preventing oil phase leakage.