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Four-Colour B-Cell ELISPOT Assay for Detection of all Four Human IgG Subclasses

CTL’s enzyme-linked immunospot (ELISPOT) assays are a key research tool for enumerating antigen-specific B-cells. Resting memory B-cells can be detected in blood after polyclonal stimulation.

In contrast, the spontaneous production of immunoglobulins (lg) by B-cells in freshly isolated blood signifies recent and ongoing antigen stimulation in-vivo.

So far, ELISPOT assays have been performed in single colours, detecting each lg class or lgG subclass in a separate assay to detect antibody-secreting cells (ASC). The downside to this traditional approach is that for a comprehensive study of all four lg classes or subclasses, four times the amount of peripheral blood mononuclear cells (PBMC), antigen and labour is needed.

CTL reports here the developments of a four-colour B-cell ELISPOT assay that simultaneously detects all lgG subclasses.

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