Supplemental web site for
single-molecule interaction profiling of DNA barcoded proteins
Chao Li2, John Aach1, David E. Hill1,3, Marc Vidal1,3 & George M. Church1,2,*
of Genetics, Harvard Medical School, Boston, MA 02115, USA.
Institute for Biologically Inspired Engineering, Harvard University, Boston, MA
for Cancer Systems Biology (CCSB) and Department of Cancer Biology, Dana-Farber
Cancer Institute, Boston, Massachusetts 02215, USA.
* To whom
correspondence should be addressed: E-mail: email@example.com (L.G.); firstname.lastname@example.org
contrast with advances in massively parallel DNA sequencing, high-throughput
protein analyses are often limited by ensemble measurements, individual analyte
purification and hence compromised quality and cost-effectiveness. Single-molecule
(SM) protein detection achieved using optical methods is limited by the number
of spectrally nonoverlapping chromophores. Here, we introduce a single
molecular interaction-sequencing (SMI-Seq) technology
for parallel protein interaction profiling leveraging SM advantages. DNA
barcodes are attached to proteins collectively via ribosome display or
individually via enzymatic conjugation. Barcoded proteins are assayed en masse
in aqueous solution and subsequently immobilized in a polyacrylamide (PAA) thin
film to construct a random SM array, where barcoding DNAs are amplified into in
situ polymerase colonies (polonies) and analyzed by DNA sequencing. This method
allows precise quantification of various proteins with an array density
theoretically higher than one million polonies per square millimeter.
Furthermore, protein interactions can be measured based on the statistics of
colocalized polonies arising from barcoding DNAs of interacting proteins. Two
demanding applications, G-protein coupled receptor (GPCR) and antibody binding profiling, were demonstrated. SMI-Seq
enables “library vs. library” screening in a one-pot assay, simultaneously
interrogating molecular binding affinity and specificity.
This web site provides access to
supplemental data files and programs related to the analysis described in the
For scripts, programs, and related
documentation, go here to accept program license
terms and access the materials.
To obtain other data being released with this
study, go here.
Please contact John Aach
with any questions or suggestions related to this web site.
Last modified: 6/23/2014 4:48 PM