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Related Concept Videos

Enzyme-Linked Immunosorbent Assay01:33

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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Development and Validation of an Ultrasensitive Single Molecule Array Digital Enzyme-linked Immunosorbent Assay for Human Interferon-α
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Single molecule enzyme-linked immunosorbent assays: theoretical considerations.

Lei Chang1, David M Rissin, David R Fournier

  • 1Quanterix Corporation, Cambridge, MA 02139, USA.

Journal of Immunological Methods
|February 29, 2012
PubMed
Summary

We developed digital ELISA, a sensitive immunoassay detecting single molecules in femtoliter wells. This method accurately predicts and measures target proteins like prostate specific antigen (PSA), enhancing assay efficiency and specificity.

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Published on: September 7, 2018

Area of Science:

  • Biochemistry
  • Analytical Chemistry
  • Molecular Biology

Background:

  • Immunoassays are crucial for detecting biomarkers.
  • Existing methods face limitations in sensitivity and efficiency.
  • Digital ELISA offers a novel approach to single-molecule detection.

Purpose of the Study:

  • To provide the theoretical basis for digital ELISA design.
  • To predict immunocomplex formation and detection using bimolecular interaction equations.
  • To validate the assay's performance with prostate specific antigen (PSA) detection.

Main Methods:

  • Development of digital ELISA utilizing single enzyme-linked immunocomplex detection.
  • Employing single molecule arrays (SiMoA) for high-sensitivity detection.
  • Utilizing theoretical equations based on bimolecular interactions to predict assay outcomes.

Main Results:

  • Digital ELISA accurately predicts immunocomplex formation and detection by SiMoA.
  • Experimental data for PSA digital ELISA favorably compare with theoretical predictions.
  • The assay demonstrates efficiency across a range of antibody affinities and on-rates.

Conclusions:

  • Digital ELISA offers high efficiency, sensitivity, and specificity.
  • The theoretical framework enables accurate prediction of assay performance.
  • This technology enhances biomarker detection and reduces reagent concentrations.