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Enzyme-Linked Immunosorbent Assay

Enzyme-Linked Immunosorbent Assay

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.
There are many different types of ELISAs, but they all involve an antibody molecule whose constant region binds an enzyme, leaving the variable region free to bind its specific antigen. Enzyme-substrate reaction allows the antigen to be visualized or quantified.

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Related Experiment Video

Updated: May 11, 2026

High-Throughput Automated Multiplex Immunofluorescence Assays for Translational Research

High-Throughput Automated Multiplex Immunofluorescence Assays for Translational Research

Published on: June 10, 2025

Multiplexed single molecule immunoassays.

David M Rissin¹, Cheuk W Kan, Linan Song

¹Quanterix Corporation, 113 Hartwell Avenue, Lexington, MA 02421, USA.

|May 31, 2013

Summary

This summary is machine-generated.

We developed a novel digital ELISA method for highly sensitive protein detection. This single-molecule counting assay achieves subfemtomolar sensitivity, significantly outperforming current multiplexed immunoassays for disease diagnostics.

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Simultaneous Detection of Different Antibody Classes in a Multiplexed Serological Test

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Area of Science:

Biochemistry
Molecular Biology
Analytical Chemistry

Background:

Multiplexed immunoassays are crucial for disease diagnostics.
Current multiplexed assays lack the sensitivity for early disease detection at low protein concentrations.

Purpose of the Study:

To develop a highly sensitive method for multiplexed protein detection.
To enable quantification of proteins at subfemtomolar concentrations using single-molecule counting.

Main Methods:

Developed a digital ELISA assay utilizing paramagnetic beads with distinct fluorescent labels.
Antibodies immobilized on beads captured specific proteins, followed by enzyme labeling.
Single molecules were detected in femtoliter wells (Single Molecule Arrays - Simoa) using fluorescence imaging.

Main Results:

Achieved single-molecule resolution for protein detection.
Simultaneously detected TNF-α, IL-6, IL-1α, and IL-1β in human plasma.
Demonstrated subfemtomolar sensitivity, 200- to 1000-fold greater than existing multiplexed immunoassays.

Conclusions:

The developed digital ELISA enables simultaneous, specific, and sensitive protein measurement.
This high-sensitivity multiplexed assay has potential for more reliable disease diagnoses.