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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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Updated: Sep 18, 2025

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

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

Published on: June 14, 2018

Label-Free Single-Molecule Immunoassay.

Xiaoyan Zhou^1,2, Chao Chen^1,3, Shuang Zhou⁴

¹Center for Bioelectronics and Biosensors, The Biodesign Institute, Arizona State University, Tempe, AZ, 85287, USA.

Advanced Science (Weinheim, Baden-Wurttemberg, Germany)

|June 20, 2025

Summary

This summary is machine-generated.

This study introduces a novel label-free single-molecule immunoassay (LFSMiA) for real-time detection of blood biomarkers. This technique offers ultra-sensitive, direct protein detection in neat blood, improving early disease diagnosis.

Keywords:

digital immunoassay label‐free plasmonic scattering microscopy single‐molecule whole blood

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Last Updated: Sep 18, 2025

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

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

Published on: June 14, 2018

Development of a Lateral Flow Immunochromatographic Strip for Rapid and Quantitative Detection of Small Molecule Compounds

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Published on: November 13, 2021

A Label-free Technique for the Spatio-temporal Imaging of Single Cell Secretions

A Label-free Technique for the Spatio-temporal Imaging of Single Cell Secretions

Published on: November 23, 2015

Area of Science:

Biomedical Engineering
Analytical Chemistry
Molecular Diagnostics

Background:

Current single-molecule immunoassays often require labeling and suffer from matrix effects and autofluorescence.
Endpoint detection limits the precision and real-time monitoring of biomarker concentrations.

Purpose of the Study:

To develop a real-time, label-free single-molecule immunoassay (LFSMiA) for ultra-sensitive biomarker detection.
To overcome limitations of existing methods, including matrix effects and the need for signal amplification.

Main Methods:

Utilized plasmonic scattering microscopy-based mass imaging for real-time, label-free detection.
Employed a 2-step sandwich assay format with dynamic tracking of single binding events.
Applied a Bayesian Gaussian process model for enhanced measurement precision.

Main Results:

Achieved sub-femtomolar detection limits and an eight-log dynamic range for biomarkers like IL-6 and PSA in various blood matrices.
Demonstrated comparable performance using an inexpensive, miniaturized setup.
Validated LFSMiA with N-terminal pro-B-type natriuretic peptide in patient samples, showing strong correlation (r > 0.99) with clinical results.

Conclusions:

LFSMiA enables ultra-sensitive, direct, and real-time protein detection at the single-molecule level in complex biological samples like neat blood.
The technology shows significant potential for early disease diagnosis and point-of-care applications due to its sensitivity, dynamic range, and translational capabilities.