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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: Sep 1, 2025

Bio-layer Interferometry for Measuring Kinetics of Protein-protein Interactions and Allosteric Ligand Effects

Bio-layer Interferometry for Measuring Kinetics of Protein-protein Interactions and Allosteric Ligand Effects

Published on: February 18, 2014

Immunoassay design and biotin interference.

Amitava Dasgupta¹

¹Department of Pathology and Laboratory Medicine, University of Kansas Medical Center, Kansas City, KS, United States.

Advances in Clinical Chemistry

|August 11, 2022

Summary

This summary is machine-generated.

High-dose biotin supplements can interfere with common lab tests, leading to diagnostic errors. This review explores biotin

Keywords:

Biotin Falsely lower troponin Hyperthyroidism Interferences Myocardial infarction

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Last Updated: Sep 1, 2025

Bio-layer Interferometry for Measuring Kinetics of Protein-protein Interactions and Allosteric Ligand Effects

Bio-layer Interferometry for Measuring Kinetics of Protein-protein Interactions and Allosteric Ligand Effects

Published on: February 18, 2014

In Vitro Biochemical Assays using Biotin Labels to Study Protein-Nucleic Acid Interactions

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Development and Validation of an Ultrasensitive Single Molecule Array Digital Enzyme-linked Immunosorbent Assay for Human Interferon-α

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Published on: June 14, 2018

Area of Science:

Biochemistry
Clinical Chemistry
Endocrinology

Background:

Biotin (vitamin B7/H) is essential for carboxylase enzymes involved in metabolism.
While beneficial for rare deficiencies, supplemental biotin offers no general health benefits.
Widespread biotin use for hair/nail health is common, despite limited evidence for efficacy in healthy individuals.

Purpose of the Study:

To review the biochemistry of biotin.
To discuss biotin's role as a potential interferent in biotin-based immunoassay formats.
To highlight the clinical implications of biotin interference in diagnostic testing.

Main Methods:

Literature review of biotin biochemistry and immunoassay interference.
Analysis of biotin's impact on competitive and sandwich immunoassay formats.
Examination of diagnostic errors caused by biotin interference, particularly in thyroid function tests.

Main Results:

Elevated biotin levels can cause falsely high results in competitive immunoassays (e.g., FT3, FT4).
Elevated biotin levels can cause falsely low results in sandwich immunoassays (e.g., TSH).
These interferences can lead to misdiagnosis, notably in hyperthyroidism evaluation.

Conclusions:

Supplemental biotin use can significantly interfere with biotin-based immunoassays.
Understanding biotin interference mechanisms is crucial for accurate laboratory diagnostics.
Clinicians should consider biotin intake when interpreting immunoassay results, especially thyroid function tests.