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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: Jun 2, 2026

Methods for Quantitative Detection of Antibody-induced Complement Activation on Red Blood Cells

Methods for Quantitative Detection of Antibody-induced Complement Activation on Red Blood Cells

Published on: January 29, 2014

Specificity of EIA immunoassay for complement factor Bb testing.

Igor Y Pavlov¹, Nikol De Forest, Julio C Delgado

¹ARUP Institute for Clinical and Experimental Pathology, Department of Pathology, University of Utah School of Medicine, Salt Lake City, Utah 84108, USA. igor.pavlov@aruplab.com

Clinical Laboratory

|April 20, 2011

Summary

This summary is machine-generated.

Measuring factor Bb levels in plasma is reliable for assessing alternative complement pathway activation. This study confirms minimal cross-reactivity with factor B, validating Bb assays for clinical use.

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Simultaneous Detection of Different Antibody Classes in a Multiplexed Serological Test

Simultaneous Detection of Different Antibody Classes in a Multiplexed Serological Test

Published on: July 14, 2023

Area of Science:

Immunology
Complement System Biology

Background:

Alternative complement pathway activation involves cleavage of factor B into Ba and Bb fragments.
Low fragment concentrations pose detection challenges due to potential cross-reactivity with intact factor B.

Purpose of the Study:

To investigate the cross-reactivity of the Quidel Bb Enzyme Immunoassay (EIA) assay.
To determine the reliability of measuring factor Bb for alternative complement pathway activation.

Main Methods:

Analyzed 109 healthy donor EDTA plasma and 80 serum samples.
Utilized factor B immunodiffusion and Quidel Bb EIA assays for sample analysis.

Main Results:

Physiological concentrations of purified factor B showed approximately 0.15% cross-reactivity in the Quidel Bb EIA assay.
Serum Bb concentrations were higher than plasma Bb, reflecting complement activation during clotting.

Conclusions:

Despite a minor 0.15% cross-reactivity, factor Bb plasma measurements are adequate for evaluating alternative complement pathway activation.
The findings support the use of Bb assays for assessing complement system status.