Abseq: Ultrahigh-throughput single cell protein profiling with droplet microfluidic barcoding.
Abseq: Ultrahigh-throughput single cell protein profiling with droplet microfluidic barcoding.
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DOI：
10.1038/srep44447
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发表时间：
2017-03-14
影响因子：
4.6
通讯作者：
Abate AR
中科院分区：
文献类型：
--
作者：
Shahi P;Kim SC;Haliburton JR;Gartner ZJ;Abate AR
蛋白质是细胞功能的主要效应物，包括细胞代谢、结构动力学和信息处理。然而，在单细胞水平的蛋白质的定量表征是具有挑战性的，由于可用的蛋白质的量很少。在这里，我们提出了Abseq，一种在单细胞中检测和定量蛋白质的方法。与流式细胞术和质谱仪一样，Abseq使用特异性抗体来检测感兴趣的表位;然而，与这些方法不同的是，抗体标记有可以用微流体条形码和DNA测序读出的序列标签。我们通过在单细胞水平上表征不同细胞类型的表面蛋白并通过其蛋白质表达谱区分细胞来证明这种新方法。DNA标记的抗体为在单细胞中分析蛋白质提供了多种优势，包括扩增低丰度标签以使其可通过测序检测的能力，使用分子指标进行定量结果，以及基本上无限的多重化。
Proteins are the primary effectors of cellular function, including cellular metabolism, structural dynamics, and information processing. However, quantitative characterization of proteins at the single-cell level is challenging due to the tiny amount of protein available. Here, we present Abseq, a method to detect and quantitate proteins in single cells at ultrahigh throughput. Like flow and mass cytometry, Abseq uses specific antibodies to detect epitopes of interest; however, unlike these methods, antibodies are labeled with sequence tags that can be read out with microfluidic barcoding and DNA sequencing. We demonstrate this novel approach by characterizing surface proteins of different cell types at the single-cell level and distinguishing between the cells by their protein expression profiles. DNA-tagged antibodies provide multiple advantages for profiling proteins in single cells, including the ability to amplify low-abundance tags to make them detectable with sequencing, to use molecular indices for quantitative results, and essentially limitless multiplexing.