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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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How to set up an ELISA.

¹Department of Immunology, Imperial College School of Medicine, London, UK.

Methods in Molecular Medicine

|February 22, 2011

Summary

Enzyme-linked immunosorbent assay (ELISA) offers sensitive molecular detection. This chapter details indirect sandwich ELISA for high sensitivity and direct ELISA for single-antibody detection.

Area of Science:

Biochemistry
Immunology
Analytical Chemistry

Background:

The enzyme-linked immunosorbent assay (ELISA) is a widely used immunoassay technique.
Numerous variations of the ELISA exist, building upon the original principle.

Purpose of the Study:

To focus on two key ELISA variations: indirect sandwich ELISA and direct ELISA.
To highlight the applications and advantages of each method.

Main Methods:

Detailed explanation of the indirect sandwich ELISA.
Description of the basic direct ELISA protocol.

Main Results:

Indirect sandwich ELISA offers high sensitivity and specificity for molecular detection.
Direct ELISA is a valuable method when only one antibody to the sample antigen is available.

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Conclusions:

ELISA is a versatile and sensitive technique for quantitative molecular detection.
Understanding the principles of indirect sandwich and direct ELISA is crucial for their effective application.