A flow-cytometry-based protocol using diverse cell types for detecting autoantibodies from human plasma and serum

Andrew Kam Ho Wong1, Deanna A Kulpa1, Guido Silvestri1

  • 1Department of Pathology and Laboratory Medicine, Emory University School of Medicine, Atlanta, GA 30322, USA.

STAR Protocols
|November 11, 2021
PubMed

Insights

This study presents a cost-effective flow cytometry protocol for detecting autoantibodies in human samples against extracellular antigens. This method offers a faster alternative to protein microarrays without needing prior target knowledge.

Area of Science:

  • Immunology
  • Biotechnology
  • Cell Biology

Background:

  • Autoantibodies play a crucial role in autoimmune diseases.
  • Current methods for autoantibody detection can be expensive and time-consuming.
  • Identifying novel autoantibodies is essential for understanding disease mechanisms.

Purpose of the Study:

  • To describe a novel, cost-effective protocol for detecting autoantibodies using flow cytometry.
  • To enable the detection of autoantibodies against a wide range of extracellular antigens.
  • To provide an accessible method for researchers without prior knowledge of potential targets.

Main Methods:

  • A cell-based flow cytometry protocol was developed for analyzing human plasma and serum samples.
  • The protocol utilizes standard flow cytometry equipment.
  • Antigen detection is limited to the cell types included in the assay.

Main Results:

  • The protocol allows for the detection of autoantibodies against numerous extracellular antigens.
  • This method is more economical and quicker than traditional protein microarrays.
  • No prior knowledge of specific autoantibody targets is required.

Conclusions:

  • This flow cytometry protocol offers a practical and efficient approach for autoantibody detection.
  • Further research is needed to correlate autoantibody detection with clinical outcomes.
  • The protocol expands the possibilities for autoantibody discovery in various research settings.

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