Single-Cell Biophysics: Measurement, Modulation, and Modeling

Single-Cell Biophysics: Measurement, Modulation, and Modeling

Poster Abstracts

128

60-POS

Board 30

Quantitative Single Analysis of C-Jun/AP-1 for Clinical Studies

Shao-Ju Lu

, Keith Willison, David Klug.

Imperial college, London, United Kingdom.

Both c-Jun and its dimerization partners in transcription factor AP-1 formation play important

roles in cell proliferation, apoptosis and cell transformation. AP-1/c-Jun is documented to be one

of the key activators in breast cancer. AP-1-mediated regulation determined cell fates critically

depends on the relative abundance of AP-1 subunits. However, bulk measurements of AP-1/c-

Jun levels do not record the intrinsic and extrinsic variation in cellular responses across a

population of cells, thereby masking the individual details.

We have developed a label-free, microfluidic antibody chip platform called the MAC chip

, to

quantify precisely the copy numbers of many proteins from a single cell. Utilizing microfluidic

strategies along with optical trapping and lysis techniques, we have established c-Jun assay

which is capable of quantification of c-Jun, phospho-c-Jun and its homodimer/ heterodimer

complex individually at the single cell level. Furthermore, the protein assays can now be

accomplished in a multiplexed single assay format.

We are working on investigations into protein noise in tumor cells isolated from biopsies of

cancer patients through the identification of biomolecular signatures.

Using patient-derived

colorectal cancer xenografts as proof-of-concept, we have demonstrated a potential route for

moving single cell proteomics into medical diagnostics. Our aim is to integrate the protein

measurements with single cell transcriptomics. In the future, we hope to gain deep insights into

c-Jun transcriptional and translational regulation at the single cell level.

1. E. Burgin, A. Salehi-Reyhani, M. Barclay, A. Brown, J. Kaplinsky, M. Novakova, M. A. A.

Neil, O. Ces, K. R. Willison, and D. R. Klug, Analyst, 2014, 139, 3235–44.

2. K. R. Willison and D. R. Klug, Curr. Opin. Biotechnol., 2013, 24, 745–51.