Mabtech AB, Box 1233, SE-13128, Nacka Strand, Sweden.
Cells. 2012 Apr 20;1(2):27-34. doi: 10.3390/cells1020027.
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.
我们评估了一种新型的 B 细胞 FluoroSpot 检测法，用于分析健康个体经肌肉注射接种甲型流感（H1N1）抗原（Pandemrix®，葛兰素史克）后的抗体反应。使用 FluoroSpot 检测法和并行运行的 ELISpot 检测法进行比较，我们在接种后 7 天测量了分泌抗原特异性以及总 IgG 或 IgA 抗体的细胞的频率。这些检测法基于高亲和力单克隆抗体，用于捕获和检测人 IgG 和 IgA。虽然传统的 ELISpot 分别分析 IgG 和 IgA 分泌 B 细胞，但荧光检测可以在同一孔中同时计数分泌 IgG 或 IgA 的 B 细胞。FluoroSpot 方案也更简单，因为该检测法无需扩增检测步骤即可进行。FluoroSpot 检测法除了具有常规 ELISpot 检测法的所有优势，包括高灵敏度、稳健性和易于操作外，还通过降低成本、时间和材料进一步增加了价值。
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