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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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Snap Chip for Cross-reactivity-free and Spotter-free Multiplexed Sandwich Immunoassays

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Cross-talk-free multiplexed immunoassay using a disposable electrochemiluminescent immunosensor array coupled with a

Cuifang Li¹, Zhifeng Fu, Zongyun Li

¹Key Laboratory of Luminescence and Real-Time Analysis (Ministry of Education), College of Pharmaceutical Sciences, Southwest University, Chongqing, China.

Biosensors & Bioelectronics

|July 23, 2011

Summary

A novel disposable electrochemiluminescent (ECL) immunosensor array was developed for multiplexed immunoassay (MIA). This method offers a sensitive, low-cost, and time-saving approach for detecting multiple analytes simultaneously.

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Area of Science:

Electrochemistry
Biosensors
Immunochemistry

Background:

Multiplexed immunoassay (MIA) is crucial for simultaneous detection of multiple analytes.
Existing MIA methods often suffer from cross-talk and complexity.
Development of sensitive and cost-effective MIA platforms is needed.

Purpose of the Study:

To develop a novel disposable electrochemiluminescent (ECL) immunosensor array for multiplexed immunoassay (MIA).
To demonstrate the feasibility of site-selective antigen immobilization and competitive immunoassay format.
To evaluate the performance for simultaneous detection of multiple immunoglobulin G (IgG) analytes.

Main Methods:

Fabrication of a disposable immunosensor array on a screen-printed carbon electrode (SPCE) substrate.
Site-selective immobilization of multiple antigens on individual working electrodes.
Competitive immunoassay using tri(2,2'-bipyridyl)ruthenium(II)-labeled antibodies.
Sequential detection of ECL signals using a photomultiplier and a custom switch to prevent cross-talk.

Main Results:

The immunosensor array successfully performed MIA for human, rabbit, and mouse IgG.
Achieved linear ranges of 10-400 ng/mL, 20-400 ng/mL, and 20-400 ng/mL, respectively.
Obtained low detection limits of 2.9, 6.1, and 6.5 ng/mL (S/N=3).

Conclusions:

The developed ECL immunosensor array offers a simple, sensitive, and cost-effective approach for MIA.
The sequential detection strategy effectively eliminates cross-talk between immunosensors.
This technology holds significant potential for point-of-care testing and field analysis of bio-agents.