PT  - JOURNAL ARTICLE
AU  - Jessie A.G. van Buggenum
AU  - Jan P. Gerlach
AU  - Sabine E.J. Tanis
AU  - Mark Hogeweg
AU  - Jesse Middelwijk
AU  - Ruud van der Steen
AU  - Cornelis A. Albers
AU  - Klaas W. Mulder
TI  - Immuno-Detection by sequencing (ID-seq) enables large-scale high-dimensional phenotyping in cells
AID  - 10.1101/158139
DP  - 2017 Jan 01
TA  - bioRxiv
PG  - 158139
4099  - http://biorxiv.org/content/early/2017/10/30/158139.short
4100  - http://biorxiv.org/content/early/2017/10/30/158139.full
AB  - 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 used ID-seq to simultaneously measure 84 (phospho-)proteins in hundreds of samples and screen the effects of ~300 kinase inhibitor probes on primary human epidermal stem cells to characterise the role of 225 kinases. Our work highlighted a previously unrecognized downregulation of mTOR signaling during differentiation and uncovered 13 kinases regulating epidermal renewal through distinct mechanisms.