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Detecting all Immunoglobulin Classes and Subclasses in a Multiplex 7 Color ImmunoSpot® Assay
Richard Caspell1, Paul V Lehmann2
1Cellular Technology Ltd., Shaker Heights, OH, USA.
Methods in Molecular Biology (Clifton, N.J.)
|June 30, 2018
Summary
Measuring memory B cells directly offers reliable immune response insights. A new multiplex assay simultaneously quantifies antigen-specific B cells across all major immunoglobulin classes, streamlining immune memory assessment.
Area of Science:
- Immunology
- Cellular Biology
Background:
- Antibody half-life in serum is short (approx. 20 days), making serum titers unreliable for assessing immunological memory.
- Memory B cells persist for decades, providing a direct measure of immune response to antigens.
- Traditional ELISPOT assays detect single immunoglobulin classes, necessitating multiple tests for comprehensive B cell memory analysis.
Purpose of the Study:
- To develop a novel multiplex assay for simultaneous enumeration of antigen-specific memory B cells across all major immunoglobulin classes and subclasses.
- To overcome the limitations of single-color ELISPOT assays for comprehensive B cell memory monitoring.
Main Methods:
- Development of a multiplex seven-color B cell ImmunoSpot assay.
- Simultaneous detection of antigen-specific B cells producing IgM, IgA, IgE, IgG1, IgG2, IgG3, and IgG4.
Main Results:
- The multiplex assay allows simultaneous quantification of antigen-specific B cells for all major antibody classes and subclasses.
- This method significantly reduces the required peripheral blood mononuclear cells (PBMCs), antigen, and labor compared to performing seven individual assays.
Conclusions:
- The developed multiplex seven-color B cell ImmunoSpot assay provides a comprehensive and efficient method for assessing B cell memory.
- This advancement enables more accurate and streamlined evaluation of long-term immune responses.
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