Cell-based small molecule screening is an effective strategy leading to new medicines. Scientists in the pharmaceutical industry as well as in academia have made tremendous progress in developing both large-scale and smaller-scale screening assays. However, an accessible and universal technology for measuring large numbers of molecular and cellular phenotypes in many samples in parallel is not available. Here we present the immuno-detection by sequencing (ID-seq) technology that combines antibody-based protein detection and DNA-sequencing via DNA-tagged antibodies. We use ID-seq to simultaneously measure 70 (phospho-)proteins in primary human epidermal stem cells to screen the effects of ~300 kinase inhibitor probes to characterise the role of 225 kinases. The results show an association between decreased mTOR signalling and increased differentiation and uncover 13 kinases potentially regulating epidermal renewal through distinct mechanisms. Taken together, our work establishes ID-seq as a flexible solution for large-scale high-dimensional phenotyping in fixed cell populations.

中文翻译：

---

通过测序进行免疫检测可以在细胞中进行大规模的高维表型分析。

基于细胞的小分子筛查是开发新药的有效策略。制药行业和学术界的科学家在开发大规模和较小规模的筛选测定法方面都取得了巨大的进步。但是，尚无可并行使用的通用技术来测量许多样品中大量的分子和细胞表型。在这里，我们介绍了通过序列免疫检测（ID-seq）技术，该技术结合了基于抗体的蛋白质检测和通过带DNA标签的抗体的DNA测序。我们使用ID-seq同时测量主要人类表皮干细胞中的70种（磷酸化）蛋白，以筛选〜300种激酶抑制剂探针的作用，以表征225种激酶的作用。结果表明，降低的mTOR信号传导与增加的分化之间存在关联，并揭示了通过独特机制潜在调节表皮更新的13种激酶。综上所述，我们的工作将ID-seq建立为灵活的解决方案，用于固定细胞群中的大规模高维表型分析。