Issue 16, 2013

Ultra-high density protein spots achieved by on chip digitalized protein synthesis

Abstract

Current methodologies for arraying proteins using cell-free protein synthesis on a chip have spatial limitations that prevent reaching ultra-high density necessary for high throughput analysis. To circumvent this, we developed an on-chip method based on microcompartmentalization of protein synthesis. Proteins are synthesized in arrayed micrometer scale chambers from confined DNA template molecules. On-chip protein expression is highly efficient and the method can be used with a minimal amount of template i.e. single DNA molecules to perform digitalized cell-free protein synthesis (d-CFPS). A functionalized surface at the floor of the tightly sealed microchambers enables direct capture of expressed proteins. A density of 104 spots per mm2 was achieved, which represents a gain by more than 3 orders of magnitude over conventional methods. This technique of forming such densely arrayed small protein spots is the first step towards the development of a general method that would allow fabrication of ultra-high density protein arrays for high-throughput analysis.

Graphical abstract: Ultra-high density protein spots achieved by on chip digitalized protein synthesis

Supplementary files

Article information

Article type
Paper
Submitted
30 Dec 2012
Accepted
15 May 2013
First published
16 May 2013

Analyst, 2013,138, 4663-4669

Ultra-high density protein spots achieved by on chip digitalized protein synthesis

S. H. Kim, S. Yoshizawa, S. Takeuchi, T. Fujii and D. Fourmy, Analyst, 2013, 138, 4663 DOI: 10.1039/C3AN00976A

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