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Related Concept Videos

Enzyme-Linked Immunosorbent Assay01:33

Enzyme-Linked Immunosorbent Assay

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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: Jul 29, 2025

Author Spotlight: Expanding the Scope of Multiplex Immunoassays for Lyme Borreliosis Diagnostics and Pathogen Research
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MultiSero: An Open-Source Multiplex-ELISA Platform for Measuring Antibody Responses to Infection.

Janie R Byrum1, Eric Waltari1, Owen Janson2,3

  • 1Chan Zuckerberg Biohub-San Francisco, San Francisco, CA 94158, USA.

Pathogens (Basel, Switzerland)
|May 27, 2023
PubMed
Summary
This summary is machine-generated.

We developed multiSero, an open-source multiplex ELISA platform for simultaneous antibody detection against multiple viral antigens. This accessible tool enhances serosurveillance studies for diseases like COVID-19.

Keywords:
ELISASARS-CoV-2multiplexopen-sourceserologyserosurveillance

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Area of Science:

  • Immunology
  • Virology
  • Biotechnology

Background:

  • Multiplexed enzyme-linked immunosorbent assays (ELISA) offer potential for expanded serosurveillance by detecting antibodies against multiple antigens simultaneously.
  • Existing multiplex ELISA platforms often lack the simplicity, robustness, and accuracy of conventional single-antigen ELISAs, limiting their widespread adoption.
  • There is a need for accessible, open-source multiplex ELISA solutions to advance the study of viral infections and immune responses.

Purpose of the Study:

  • To develop and validate multiSero, an open-source multiplex ELISA platform for measuring antibody responses to viral infections.
  • To assess the performance of multiSero in terms of sensitivity, specificity, and correlation with existing diagnostic methods.
  • To demonstrate the utility of multiSero for tracking antibody dynamics in response to vaccination.

Main Methods:

  • Development of a three-part multiplex ELISA system: (1) ELISA against a protein array in a 96-well format, (2) automated imaging using an open-source plate reader, and (3) automated optical density measurement via an open-source analysis pipeline.
  • Validation using 217 human serum samples, comparing antibody binding to Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) antigens.
  • Correlation analysis of multiSero-determined antibody titers with commercially available SARS-CoV-2 antibody tests.

Main Results:

  • The multiSero platform demonstrated high diagnostic accuracy for classifying seropositivity, with sensitivity of 0.978 and specificity of 0.977.
  • A strong correlation was observed between antibody titers determined by multiSero and those measured by commercial SARS-CoV-2 antibody tests.
  • Antigen-specific changes in antibody titer dynamics were successfully identified following vaccination, highlighting the platform's ability to track immune responses.

Conclusions:

  • The open-source multiSero platform provides a simple, robust, and accurate method for multiplexed antibody detection.
  • MultiSero's accessibility and performance make it a valuable tool for advancing serosurveillance studies for SARS-CoV-2 and other significant viral pathogens.
  • The platform facilitates detailed analysis of antibody responses, aiding in the understanding of infection and vaccination impacts.