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Cells 2012, 1(2), 27-34; doi:10.3390/cells1020027

Comparison of ELISpot and FluoroSpot in the Analysis of Swine Flu-Specific IgG and IgA Secretion by in Vivo Activated Human B Cells

Mabtech AB, Box 1233, SE-13128, Nacka Strand, Sweden
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Received: 13 March 2012 / Revised: 4 April 2012 / Accepted: 6 April 2012 / Published: 20 April 2012
(This article belongs to the Special Issue ELISPOT Research)
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Abstract

We have evaluated a novel B-cell FluoroSpot assay for the analysis of antibody responses in healthy individuals vaccinated intramuscularly with Influenza A (H1N1) antigen (Pandemrix®, GlaxoSmithKline). Using the FluoroSpot assay and an ELISpot assay run in parallel for comparison, we measured the frequency of cells secreting antigen-specific as well as total IgG or IgA antibodies seven days post vaccination. The assays were based on high affinity monoclonal antibodies for capture and detection of human IgG and IgA. Whereas conventional ELISpot analyzes IgG- and IgA-secreting B cells separately, fluorescent detection enabled simultaneous enumeration of B cells secreting IgG or IgA in the same well. The FluoroSpot protocol was also simpler as the assay could be performed without the need for an amplifying detection step. While having all the advantages of a conventional ELISpot assay, including high sensitivity, robustness and ease of performance, the FluoroSpot assay adds further value in reducing costs, time and material. View Full-Text
Keywords: Influenza A (H1N1); vaccination; B cells; IgG and IgA secretion; FluoroSpot; ELISpot Influenza A (H1N1); vaccination; B cells; IgG and IgA secretion; FluoroSpot; ELISpot
This is an open access article distributed under the Creative Commons Attribution License (CC BY 3.0).

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MDPI and ACS Style

Kesa, G.; Larsson, P.H.; Ahlborg, N.; Axelsson, B. Comparison of ELISpot and FluoroSpot in the Analysis of Swine Flu-Specific IgG and IgA Secretion by in Vivo Activated Human B Cells. Cells 2012, 1, 27-34.

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