Department of Bioengineering and Therapeutic Sciences and California Institute for Quantitative Biosciences (QB3), University of California, San Francisco, San Francisco, CA 94158, USA.
Department of Pharmaceutical Chemistry, University of California, San Francisco, San Francisco, CA 94158, USA.
Sci Rep. 2017 Mar 14;7:44447. doi: 10.1038/srep44447.
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.
蛋白质是细胞功能的主要效应物，包括细胞代谢、结构动态和信息处理。然而，由于可获得的蛋白质数量极少，因此在单细胞水平上对蛋白质进行定量表征具有挑战性。在这里，我们提出了 Abseq，这是一种在超高通量下检测和定量单细胞中蛋白质的方法。与流式细胞术和质谱细胞术一样，Abseq 使用特定的抗体来检测感兴趣的表位；然而，与这些方法不同的是，抗体被带有序列标签的标签标记，这些标签可以通过微流控条码和 DNA 测序读出。我们通过在单细胞水平上表征不同细胞类型的表面蛋白，并根据其蛋白表达谱来区分细胞，证明了这种新方法。DNA 标记的抗体为在单细胞中分析蛋白质提供了多个优势，包括能够扩增低丰度标签以使其可通过测序检测、使用分子指标进行定量结果以及实质上无限制的多重检测。
