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Antibody Surface Coverage Drives Matrix Interference in Microfluidic Capillary Immunoassays.

Ana I Barbosa1,2, Alexander D Edwards2,3, Nuno M Reis2,4

  • 1Department of Chemical Engineering, Loughborough University, Loughborough LE11 3TU, United Kingdom.

ACS Sensors
|June 17, 2021
PubMed
Summary

Optimizing biosensor antibody surface coverage in serum, not buffer, minimizes sample interference. This novel approach enhances biosensor development for point-of-care diagnostics without added complexity or cost.

Keywords:
biosensorsmatrix effectmicrocapillary filmmicrofluidicsprotein biomarkers

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Area of Science:

  • Biomedical Engineering
  • Analytical Chemistry
  • Biosensor Technology

Background:

  • Biosensor performance is often inconsistent when moving from buffer optimization to real biological samples.
  • Existing methods to reduce sample matrix interference are complex, hindering automation and miniaturization.

Purpose of the Study:

  • To systematically analyze matrix interference in microfluidic immunoassay systems.
  • To identify key factors influencing serum matrix effects in biosensors.
  • To develop a novel strategy for mitigating matrix interference in biosensor applications.

Main Methods:

  • Systematic analysis of hundreds of microfluidic immunoassay fluoropolymer strips.
  • Exploration of matrix interference across diverse enzymatic immunoassays (direct, sandwich assays for PSA and IL-1β).
  • Investigation of antibody surface coverage and capillary diameter effects on serum matrix interference.

Main Results:

  • Serum matrix interference is significantly influenced by antibody surface coverage on microfluidic capillaries.
  • Low-affinity serum components (e.g., autoantibodies) competing for immobilized antibodies are identified as a primary cause of matrix effects.
  • Optimizing antibody surface coverage and incubation time in serum, rather than buffer, effectively manages matrix interference.

Conclusions:

  • Antibody surface coverage is a critical parameter for managing serum matrix interference in microfluidic biosensors.
  • Optimizing bioassays directly in serum offers a cost-effective and efficient method to overcome matrix effects.
  • This approach facilitates the development of advanced point-of-care tests and biosensors.