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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...

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Related Experiment Video

Updated: Apr 7, 2026

Electrochemiluminescence Assays for Human Islet Autoantibodies

Electrochemiluminescence Assays for Human Islet Autoantibodies

Published on: March 23, 2018

Direct ELISA.

¹Produce Safety and Microbiology Unit (PSM), Western Regional Research Center (WRRC), Pacific West Area (PWA), Agricultural Research Service (ARS), United States Department of Agriculture (USDA), 800 Buchanan St., Albany, CA, 94710, USA, Alice.lin@ars.usda.gov.

Methods in Molecular Biology (Clifton, N.J.)

|July 11, 2015

Summary

This summary is machine-generated.

Enzyme-linked immunosorbent assay (ELISA) offers rapid and sensitive antigen detection. This guide details direct ELISA methods using horseradish peroxidase and enhanced chemiluminescence for optimized results.

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Area of Science:

Biochemistry
Immunology
Analytical Chemistry

Background:

Enzyme-linked immunosorbent assay (ELISA) is a key technique for antigen detection and quantification.
ELISA is widely used in clinical diagnostics and the food industry.
The technology has evolved beyond traditional plates to automated platforms.

Purpose of the Study:

To describe the steps for direct ELISA using a horseradish peroxidase (HRP)-conjugated antibody.
To detail the use of luminol-based enhanced chemiluminescence (ECL) substrate for antigen detection.
To provide a method for optimizing ELISA assays via chessboard titration.

Main Methods:

Direct ELISA protocol for plate-bound antigen detection.
Utilizing HRP-conjugated antibodies and ECL substrate.
Chessboard titration for assay optimization.

Main Results:

Detailed procedural steps for direct ELISA.
Demonstration of enhanced chemiluminescence detection.
Methodology for optimizing assay sensitivity and specificity.

Conclusions:

Direct ELISA with HRP and ECL is an effective method for antigen detection.
Assay optimization through chessboard titration is crucial for reliable results.
ELISA remains a versatile and evolving diagnostic tool.