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3D-printed capillaric ELISA-on-a-chip with aliquoting.

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Lab on a Chip
|February 1, 2023
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Summary

This study introduces a new ELISA-on-a-chip that simplifies sample loading and pipetting for high-sensitivity SARS-CoV-2 detection. The device offers accurate aliquoting and outperforms rapid tests for point-of-care diagnostics.

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Area of Science:

  • Biotechnology
  • Microfluidics
  • Assay Development

Background:

  • Miniaturized sandwich immunoassays like ELISA are valuable but often require complex equipment.
  • Existing lab-on-a-chip ELISA formats face limitations in pipetting precision and surfactant tolerance.
  • Previous microfluidic ELISA approaches, such as microfluidic chain reaction, have shown promise but require further optimization for broader adoption.

Purpose of the Study:

  • To develop an automated, user-friendly ELISA-on-a-chip with integrated aliquoting functionality.
  • To enhance the robustness of lab-on-a-chip ELISA by accommodating higher surfactant concentrations and preventing undesired mixing.
  • To demonstrate a high-sensitivity quantitative assay for SARS-CoV-2 nucleocapsid protein in diluted saliva.

Main Methods:

  • A 3D-printed ELISA-on-a-chip device with a capillaric aliquoting circuit for precise reagent and sample volumes.
  • Integration of barrier channels to control solution flow and prevent premature mixing.
  • A modular assay design with a click-connectable module containing capture antibodies and a capillary pump.
  • Quantitative readout via digitized colorimetric signal on a nitrocellulose strip.

Main Results:

  • The device achieved >93% accuracy in autonomous aliquoting of reagents and sample.
  • The assay successfully quantified SARS-CoV-2 nucleocapsid protein in 4×-diluted saliva with a limit of detection of 91 pg mL⁻¹.
  • The system demonstrated a limit of detection of 54 pg mL⁻¹ in buffer, significantly outperforming rapid tests.
  • The entire assay was completed in 1.5 hours with simplified chip loading and pipetting.

Conclusions:

  • The developed ELISA-on-a-chip offers a simplified, automated, and high-sensitivity platform for immunoassays.
  • Its design overcomes limitations of previous microfluidic ELISA systems, including precise pipetting and surfactant tolerance.
  • The technology holds potential for both automated laboratory ELISA and point-of-care diagnostic applications, combining the convenience of rapid tests with the performance of central laboratory assays.