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Related Experiment Video

Updated: Dec 27, 2025

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

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Customizable Multiplex Antibody Array Immunoassays with Attomolar Sensitivities.

Carmen I Tobos¹, Antony J Sheehan², David C Duffy¹

¹Quanterix Corporation, 900 Middlesex Turnpike, Building 1, Billerica, Massachusetts 01821, United States.

Analytical Chemistry

|March 4, 2020

Summary

This summary is machine-generated.

A new customizable multiplex immunoassay enables simultaneous measurement of up to five analytes with attomolar sensitivity. This enzyme-linked immunosorbent assay (ELISA) offers ultrasensitive protein detection for custom assay development and diagnostics.

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Last Updated: Dec 27, 2025

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Area of Science:

Biotechnology
Assay Development
Immunology

Background:

Traditional enzyme-linked immunosorbent assays (ELISA) often require specific capture antibodies for each target analyte.
Multiplex assays enable simultaneous detection of multiple analytes but can be limited by customization and sensitivity.
Existing methods for antibody printing in ELISA may require specialized precision instrumentation.

Purpose of the Study:

To develop a customizable, contact-printed multiplex immunoassay with attomolar sensitivity.
To create an enzyme-linked immunosorbent assay (ELISA) workflow using anchor antibodies and peptide coupling for enhanced flexibility.
To demonstrate the utility of this novel approach for simultaneous detection of multiple cytokines.

Main Methods:

Developed a contact-printed multiplex immunoassay utilizing unique anchor antibodies spotted in a circular pattern within microtiter plate wells.
Coupled specific peptides to unique assay capture antibodies, enabling a flexible multiplex immunoassay workflow.
Created a 5-plex assay to measure interleukin-5 (IL-5), IL-6, IL-10, IL-22, and tumor necrosis factor-alpha (TNF-α) in biological samples.

Main Results:

Achieved attomolar sensitivity for simultaneous measurement of up to five analytes.
Demonstrated strong correlation (r² values ranging from 0.75 to 0.99) between the novel anchor antibody method and traditional capture antibody printing for IL-5, IL-6, IL-10, IL-22, and TNF-α.
Validated the assay's performance in serum and plasma samples.

Conclusions:

The developed customizable contact-printed multiplex immunoassay provides ultrasensitive detection of multiple analytes.
This approach offers a flexible and accessible platform for custom assay development, antibody screening, and biomarker detection.
The technology is valuable for protein profiling in diagnostic applications, even without access to precision printing equipment.