title: 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.
authors.affiliation: Institute of Industrial Science, The University of Tokyo, 4-6-1, Komaba, Meguro-ku, Tokyo 153-8505, Japan.
authors.forename: Soo Hyeon
authors.initials: SH
authors.lastname: Kim
authors.forename: Satoko
authors.initials: S
authors.lastname: Yoshizawa
authors.forename: Shoji
authors.initials: S
authors.lastname: Takeuchi
authors.forename: Teruo
authors.initials: T
authors.lastname: Fujii
authors.forename: Dominique
authors.initials: D
authors.lastname: Fourmy
mesh_terms: D002474:Cell-Free System / Q000737:chemistry / Q000378:metabolism; D040081:Protein Array Analysis / Q000379:methods*; D014176:Protein Biosynthesis