European Molecular Biology Laboratory (EMBL), Genome Biology Unit, Meyerhofstrasse 1, Heidelberg, Germany.
European Molecular Biology Laboratory (EMBL), Genome Biology Unit, Meyerhofstrasse 1, Heidelberg, Germany.
Cell Rep. 2018 Feb 20;22(8):2206-2215. doi: 10.1016/j.celrep.2018.01.071.
Monoclonal antibodies are a main player in modern drug discovery. Many antibody screening formats exist, each with specific advantages and limitations. Nonetheless, it remains challenging to screen antibodies for the binding of cell-surface receptors (the most important class of all drug targets) or for the binding to target cells rather than purified proteins. Here, we present a high-throughput droplet microfluidics approach employing dual-color normalized fluorescence readout to detect antibody binding. This enables us to obtain quantitative data on target cell recognition, using as little as 33 fg of IgG per assay. Starting with an excess of hybridoma cells releasing unspecific antibodies, individual clones secreting specific binders (of target cells co-encapsulated into droplets) could be enriched 220-fold after sorting 80,000 clones in a single experiment. This opens the way for therapeutic antibody discovery, especially since the single-cell approach is in principle also applicable to primary human plasma cells.
单克隆抗体是现代药物发现的主要参与者。有许多抗体筛选格式，每种格式都有其特定的优点和局限性。尽管如此，筛选与细胞表面受体（所有药物靶点中最重要的一类）结合的抗体，或者筛选与靶细胞而不是纯化蛋白结合的抗体仍然具有挑战性。在这里，我们提出了一种高通量液滴微流控方法，采用双色归一化荧光读数来检测抗体结合。这使我们能够使用每个测定仅 33 fg 的 IgG 获得关于靶细胞识别的定量数据。从过量释放非特异性抗体的杂交瘤细胞开始，在单个实验中对 80,000 个克隆进行分选后，能够富集 220 倍分泌特异性结合物（共包裹在液滴中的靶细胞）的单个克隆。这为治疗性抗体的发现开辟了道路，特别是因为单细胞方法原则上也适用于原代人血浆细胞。
