Search research articles

ABOUT JoVE

Overview Leadership Blog JoVE Help Center

AUTHORS

Publishing Process Editorial Board Scope & Policies Peer Review FAQ Submit

LIBRARIANS

Testimonials Subscriptions Access Resources Library Advisory Board FAQ

RESEARCH

JoVE Journal Methods Collections JoVE Encyclopedia of Experiments Archive

EDUCATION

JoVE Core JoVE Business JoVE Science Education JoVE Lab Manual Faculty Resource Center Faculty Site

Terms & Conditions of Use

Related Concept Videos

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...

You might also read

Related Articles

Articles linked to this work by shared authors, journal, and citation graph.

Sort by

Same author

OspA antibodies inhibit the <i>in vitro</i> transmigration of <i>Borrelia burgdorferi</i>.

Infection and immunity·2026

Same author

Broadly conserved protective epitopes on the lyme disease vaccine antigen, OspA.

PLoS pathogens·2026

Same author

An Fc-silent OspA monoclonal antibody passively protects mice from tick and intradermal Borrelia burgdorferi challenge.

PloS one·2026

Same author

Maternal expression and breast milk transfer of an mRNA- encoded monoclonal antibody in a murine model of cholera.

PLoS neglected tropical diseases·2026

Same author

A Chip-Based Surveillance Platform for Detecting Antibody-Based Immunity and Immune Escape Using SARS-CoV-2 Pseudovirus Surrogates.

Biotechnology and bioengineering·2025

Same author

Germline encoded residues dominate the interaction of a human monoclonal antibody with decorin binding protein A of <i>Borrelia burgdorferi</i>.

Frontiers in immunology·2025

Search research articles

Related Experiment Video

Updated: Sep 17, 2025

Peptide Scanning-assisted Identification of a Monoclonal Antibody-recognized Linear B-cell Epitope

Peptide Scanning-assisted Identification of a Monoclonal Antibody-recognized Linear B-cell Epitope

Published on: March 24, 2017

B Cell Epitope Mapping by High-Density Competition ELISAs.

David J Vance¹, Nicholas J Mantis²

¹Division of Infectious Disease, New York State Department of Health, Wadsworth Center, Albany, NY, USA. David.Vance@health.ny.gov.

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

|July 2, 2025

Summary

Competition ELISAs rapidly bin monoclonal (MAb) and single-domain (V_HH) antibodies against protein antigens. This method offers an efficient alternative to resource-intensive epitope mapping techniques.

Keywords:

Antibodies Competition ELISA Epitope Nanobodies

More Related Videos

Single-cell Screening Method for the Selection and Recovery of Antibodies with Desired Specificities from Enriched Human Memory B Cell Populations

Single-cell Screening Method for the Selection and Recovery of Antibodies with Desired Specificities from Enriched Human Memory B Cell Populations

Published on: August 22, 2019

A High Throughput MHC II Binding Assay for Quantitative Analysis of Peptide Epitopes

A High Throughput MHC II Binding Assay for Quantitative Analysis of Peptide Epitopes

Published on: March 25, 2014

Related Experiment Videos

Last Updated: Sep 17, 2025

Peptide Scanning-assisted Identification of a Monoclonal Antibody-recognized Linear B-cell Epitope

Peptide Scanning-assisted Identification of a Monoclonal Antibody-recognized Linear B-cell Epitope

Published on: March 24, 2017

Single-cell Screening Method for the Selection and Recovery of Antibodies with Desired Specificities from Enriched Human Memory B Cell Populations

Single-cell Screening Method for the Selection and Recovery of Antibodies with Desired Specificities from Enriched Human Memory B Cell Populations

Published on: August 22, 2019

A High Throughput MHC II Binding Assay for Quantitative Analysis of Peptide Epitopes

A High Throughput MHC II Binding Assay for Quantitative Analysis of Peptide Epitopes

Published on: March 25, 2014

Area of Science:

Immunology
Biochemistry
Structural Biology

Background:

Large-scale discovery of monoclonal (MAb) and single-domain (V_HH) antibodies against protein antigens requires efficient methods for antibody characterization.
Traditional epitope mapping techniques like cryogenic electron microscopy (EM) and X-ray crystallography are resource-intensive.

Purpose of the Study:

To describe the application of competition enzyme-linked immunosorbent assays (ELISAs) for rapid epitope binning of antibodies.
To present alternative competition ELISA strategies for improved antigen handling and reduced artifacts.

Main Methods:

Competition ELISAs were employed to compare the binding of query antibodies to immobilized antigens in the presence of known competitor antibodies.
Alternative strategies involving capture of soluble antigen, with and without biotin tagging, were explored.
Detection utilized enzyme-linked, species-specific secondary antibodies and colorimetric substrates.

Main Results:

Competition ELISAs provide a means to rapidly bin antibodies based on their epitopes.
Alternative ELISA formats address limitations of indirect competition ELISAs, such as antigen immobilization artifacts.
Optimized competition ELISAs can generate high-resolution epitope maps from large antibody collections.

Conclusions:

Competition ELISAs are a valuable tool for high-throughput antibody screening and epitope binning.
Modified competition ELISA formats enhance reliability and applicability for large-scale antibody characterization.
This approach facilitates efficient epitope mapping, aiding in antibody discovery and development.