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Multiplex bead binding assays using off-the-shelf components and common flow cytometers.

Takamitsu Hattori1, Akiko Koide2, Tatyana Panchenko3

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Journal of Immunological Methods
|December 28, 2020
PubMed
Summary

We developed a low-cost, high-throughput multiplex bead binding assay (MBBA) using standard cytometers. This method enables rapid protein-ligand interaction analysis and reduces reagent costs, making MBBAs more accessible for research.

Keywords:
Affinity determinationAntibody screeningAntibody-antigen interactionMultiplex assayPhage display

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Area of Science:

  • Biochemistry
  • Immunology
  • Biotechnology

Background:

  • Quantifying protein-ligand interactions is crucial in biology and medicine.
  • Multiplex bead binding assays (MBBAs) enable simultaneous analysis of multiple interactions.
  • Existing MBBA platforms often require specialized equipment and reagents, limiting accessibility.

Purpose of the Study:

  • To develop a cost-effective and accessible MBBA method.
  • To enable rapid experimentation without specialized instruments.
  • To facilitate high-throughput characterization of biomolecular interactions.

Main Methods:

  • Utilized the biotin-streptavidin interaction for bead conjugation.
  • Created fluorescently labeled streptavidin-coated magnetic beads using a biotin-conjugated dye.
  • Employed standard cytometers for data acquisition.

Main Results:

  • Demonstrated a novel MBBA method using low-cost reagents and standard cytometers.
  • Successfully characterized phage-displayed antibodies against SARS-CoV-2 antigens.
  • Achieved significantly improved throughput and reduced antigen consumption compared to phage ELISA.

Conclusions:

  • The developed MBBA method offers a broadly accessible, cost-effective, and high-throughput solution for analyzing protein-ligand interactions.
  • This approach simplifies experimental procedures and reduces resource requirements.
  • The method has significant implications for antibody discovery and characterization in research and development.