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A Simple Cell-based Immunofluorescence Assay to Detect Autoantibody Against the N-Methyl-D-Aspartate NMDA Receptor in Blood
Published on: January 9, 2018
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.
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.
Abstract:
Here, we describe a protocol for cell-based detection of autoantibodies from human plasma and serum samples using a standard flow cytometer. The protocol allows detection of autoantibodies against a wide array of extracellular antigens. Antigen coverage is limited to the cell types tested, and researchers will need to further determine if autoantibody-positive samples correlate with cytotoxic or clinical outcomes. This protocol is less expensive and faster to perform when compared to protein microarrays and requires no prior knowledge of potential targets. For complete details on the use and execution of this protocol, please refer to Wong et al. (2021).
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