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Multiplex and Spatiotemporal Detection of Single-Cell Antibody Secretions Using Protein-Patterned Microwells.

Julie Van Lent1,2, Karen Ven1,2, Maya Imbrechts3,4

  • 1Department of Biosystems, Biosensors Group, KU Leuven, Leuven 3001, Belgium.

Analytical Chemistry
|October 13, 2025
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Summary

This study introduces a microfluidic device for analyzing antibody-secreting cells. The technology enables simultaneous, time-resolved screening of antibodies against multiple targets, advancing cell-based discovery.

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Area of Science:

  • Biotechnology
  • Microfluidics
  • Immunology

Background:

  • Antibodies are crucial for therapeutics and diagnostics.
  • Current single-cell technologies lack temporal and multiplex screening capabilities.
  • Discovering novel antibodies requires efficient screening of antibody-expressing cells.

Purpose of the Study:

  • To develop a microfluidic device for in-depth analysis of antibody-secreting cells.
  • To enable combined temporal and multiplex screening of antibody secretion.
  • To overcome limitations of existing single-cell analysis methods.

Main Methods:

  • Utilized a microwell-based microfluidic device.
  • Micropatterned proteins of interest onto specific regions within wells.
  • Analyzed secretion profiles of single antibody-secreting cells over time.
  • Simultaneously identified cells secreting antibodies against different proteins.

Main Results:

  • Demonstrated temporal investigation of single-cell secretion profiles.
  • Successfully identified hybridomas secreting antibodies against SARS-CoV-2 receptor binding domain and nucleocapsid simultaneously.
  • Showcased the device's capability for multiplex antibody screening.

Conclusions:

  • The developed microfluidic device enables advanced analysis of antibody-secreting cells.
  • The strategy is adaptable for various secretome studies, including ligand and cell-adhesive protein patterning.
  • This technology holds significant promise for fundamental research and drug discovery.