Anti-nuclear antibody (ANA) detection in culture supernatant of cloned B cells

Haque, S.; Watanabe, M.; Atisha-Fregoso, Y.; Diamond, B.

doi:10.1101/2023.05.12.540540

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bioRxiv · 10.1101/2023.05.12.540540

Anti-nuclear antibody (ANA) detection in culture supernatant of cloned B cells

Haque, S.·Watanabe, M.·Atisha-Fregoso, Y.·Diamond, B.

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Abstract

Anti-nuclear antibodies (ANA) of individuals with SLE contribute to disease pathogenesis and tissue injury. Systematic analysis of the fine specificity of ANA produced by individual B cells has been limited, in part, to the low concentration of immunoglobulin in single B cell cultures. We modified single B cell culture methods and developed an ANA ELISA to detect ANA at very low levels (15.0 ng/ml) of IgM in the supernatant of clonally expanded B cell cultures. This approach allows for the analysis of ANA reactivity of the IgM produced by approximately 60% of the single sorted naive B cells expanded in culture and permits a relatively high throughput evaluation at the single cell level of the ANA reactivity of naive B cells of healthy individuals and patients with autoimmune diseases.

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Haque, S., Watanabe, M., Atisha-Fregoso, Y., Diamond, B.. 2023-05-14. Anti-nuclear antibody (ANA) detection in culture supernatant of cloned B cells. https://doi.org/10.1101/2023.05.12.540540

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