Studies of the dynamics of the antibody-mediated immune response have been hampered by the absence of quantitative, high-throughput systems to analyze individual antibody-secreting cells. Here we describe a simple microfluidic system, DropMap, in which single cells are compartmentalized in tens of thousands of 40-pL droplets and analyzed in two-dimensional droplet arrays using a fluorescence relocation-based immunoassay. Using DropMap, we characterized antibody-secreting cells in mice immunized with tetanus toxoid (TT) over a 7-week protocol, simultaneously analyzing the secretion rate and affinity of IgG from over 0.5 million individual cells enriched from spleen and bone marrow. Immunization resulted in dramatic increases in the range of both single-cell secretion rates and affinities, which spanned at maximum 3 and 4 logs, respectively. We observed differences over time in dynamics of secretion rate and affinity within and between anatomical compartments. This system will not only enable immune monitoring and optimization of immunization and vaccination protocols but also potentiate antibody screening.

中文翻译：

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IgG分泌细胞的单细胞深表型分析，用于高分辨率免疫监测。

由于缺乏用于分析单个抗体分泌细胞的定量，高通量系统，因此阻碍了抗体介导的免疫反应动力学的研究。在这里，我们描述了一个简单的微流体系统DropMap，其中单个细胞在成千上万的40 pL液滴中分隔，并使用基于荧光重定位的免疫测定法在二维液滴阵列中进行分析。使用DropMap，我们在经过7周的方案的破伤风类毒素（TT）免疫的小鼠中表征了抗体分泌细胞，同时分析了从脾脏和骨髓中富集的超过50万个单个细胞中IgG的分泌率和亲和力。免疫导致单细胞分泌率和亲和力范围的急剧增加，分别跨越最大3和4个对数。我们观察到随着时间的变化，在解剖间隔内和之间的分泌速率和亲和力的动态变化。该系统不仅可以进行免疫监测以及优化免疫和接种方案，还可以增强抗体筛选。